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Pituitary Iunctinn and mnrphn!

ngy
in dngs with primary hypnthyrnidism






Maria Mnntscrrat Diaz Espincira

2008


Voor Andre, lrun, Nurlu en MlgueI

Pituitary Iunctinn and mnrphn!ngy
in dngs with primary hypnthyrnidism


Functic cn mnrIn!ngic van dc hypnIysc
bi| hnndcn mct primairc hypnthyrcndic

(nci ccn sancntaiiing in nci Nc!cr|an!s)




PrncIschriIt

ler verkri|ging van de graad van doclor aan de Universileil Ulrechl o gezag van
de reclor magnificus, rof.dr. }.C.Sloof, ingevoIge hel besIuil van hel coIIege van
romolies in hel oenbaar le verdedigen o donderdag 18 december 2008
des middags le 12:45 uur

door

Maria Mnntscrrat Diaz Espincira

geboren o 27 |uIi 1966, II IerroI (San|e)
Iromolor: Irof.dr. A. Ri|nberk

Co-romoloren: Dr. H.S. Kooislra
Dr. }.A. MoI














The sludies in lhis lhesis vere conducled al and financiaIIy suorled by lhe
Dearlmenl of CIinicaI Sciences of Comanion AnimaIs, IacuIly of Velerinary
Medicine, Ulrechl Universily, Ulrechl, The NelherIands








IubIicalion of lhis lhesis vas made ossibIe by lhe generous suorl of


lntertet Scherlng-PIough AnlmuI HeuIth

].L. ]urrluunse Stlchtlng

Cnvcr: Nuria Bncrsma
I!!ustratinns: Yvnnnc Pn!!ak
Druk: Drukkcri| Grncn, Lcidcn










Diaz Isieira, Maria Monlserral



Iiluilary funclion and morhoIogy in dogs vilh rimary hyolhyroidism
Maria Monlserral Diaz Isieira, Ulrechl
Universileil Ulrechl, IacuIleil Diergeneeskunde
Thesis Universileil Ulrechl.- Wilh ref.- Wilh summary in Dulch
ISN: 978-90-393-4947-2
Key vords: hyolhyroidism, dogs, iluilary gIand, TSH, lhyroxine,
lransdifferenlialion, lhyrolroes, GH, lhyroid
Cnntcnts

Chaptcr 1 Aims and scoe of lhe lhesis 9

Chaptcr 2 GeneraI inlroduclion 13

Chaptcr 3 Assessmenl of lhyroid funclion in dogs 49
vilh Iov Iasma lhyroxine concenlralion

Chaptcr 4 Secrelion allern of lhyroid-slimuIaling hormone 69
in dogs during eulhyroidism and hyolhyroidism

Chaptcr 5 Irimary hyolhyroidism in dogs is associaled 85
vilh eIevaled grovlh hormone reIease

Chaptcr 6 Thyrolroin-reIeasing hormone-induced 101
grovlh hormone secrelion in dogs vilh
rimary hyolhyroidism

Chaptcr 7 IunclionaI and morhoIogicaI changes in 115
lhe adenohyohysis of dogs vilh induced
rimary hyolhyroidism: Ioss of TSH
hyersecrelion, hyersomalolroism,
hyoroIaclinemia, and iluilary enIargemenl
vilh lransdifferenlialion

Chaptcr 8 AdenohyohyseaI funclion in dogs vilh 145
rimary hyolhyroidism and non-lhyroidaI iIIness

Chaptcr 9 Summarizing discussion and concIusions 165

Chaptcr 10 Samenvallende besreking en concIusies 179

Dankwnnrd 195

Curricu!um vitac 200

List nI pub!icatinns 201

List nI abbrcviatinns 203


Chaptcr 1

Aims and scnpc nI thc thcsis






Chaler 1
_________________________________________________________________________________

10

Aims and scoe of lhe lhesis
_________________________________________________________________________________
11

Thyroid hormones have effecl on aImosl every organ syslem in lhe body.
This exIains lhe vide range of cIinicaI manifeslalions of rimary hyolhyroidism
in dogs. The signs and symloms are oflen nonsecific and lhe disease onsel is
insidious. Il may lherefore lake a Iong lime before hyolhyroidism is diagnosed.
On lhe olher hand, canine hyolhyroidism is frequenlIy diagnosed erroneousIy
because many non-lhyroidaI faclors (e.g. drug lheray, concurrenl disease) may
resuIl in a Iov Iasma lolaI lhyroxine (TT4) concenlralion.
The inlroduclion of homoIogous immunoassays for measuring canine
Iasma lhyroid-slimuIaling hormone (TSH) concenlralions in lhe 1990s enabIed lhe
assessmenl of lhe canine iluilary-lhyroid axis by measuring lhe hormone air T4
and TSH, vilh lhe diagnosis of rimary hyolhyroidism being eslabIished on lhe
basis of a Iov Iasma TT4 concenlralion and a high Iasma TSH concenlralion.
Hovever, in sludies using lhe TSH-slimuIalion lesl as goId slandard, il aeared
lhal Iasma TSH concenlralions vere nol eIevaled in as many as a lhird of dogs
vilh rimary hyolhyroidism.
The main aim of lhe sludies described in lhis lhesis vas lo lesl lhe
hyolhesis lhal rimary hyolhyroidism Ieads lo adenohyohyseaI aIleralions
lhal may exIain lhe occurrence of Iov Iasma TSH concenlralions. Il vas hoed
lhal lhese sludies vouId aIso bring aboul beller vays lo diagnose rimary
hyolhyroidism in dogs.
In lhe generaI inlroduclion of lhis lhesis (Chaptcr 2), an overviev is given
of 1) deveIomenl and analomy of lhe lhyroid gIand, 2) lhyroid hysioIogy, 3)
hyolhyroidism, 4) nomencIalure of lhe iluilary gIand, 5) funclionaI asecls of
lhe iluilary gIand, and 6) uIlradian uIsaliIe secrelion of adenohyohyseaI
hormones.
The Iack of secificily of lhe T4 assay and lhe Iack of sensilivily of TSH
assay may make il difficuIl lo differenliale belveen hyolhyroidism and non-
lhyroidaI iIIness in dogs. Chaptcr 3 describes lhe diagnoslic vaIue of 1) circuIaling
concenlralions of TT4, free T4, and TSH, 2) a TSH-slimuIalion lesl, 3) Iasma TSH
concenlralions afler slimuIalion vilh TSH-reIeasing hormone (TRH), 4) circuIaling
lhyrogIobuIin anlibodies (TgAbs), and 5) lhyroidaI
99m
TcO4
-
ulake, in
dislinguishing dogs vilh hyolhyroidism and dogs vilh non-lhyroidaI iIIness. The
finaI diagnosis in lhese dogs vas based uon hisloalhoIogic examinalion of
lhyroid lissue oblained by surgicaI biosy.
To delermine vhelher lhe occurrence of a Iov Iasma TSH concenlralion
in dogs vilh rimary hyolhyroidism may be due lo fIuclualions in lhe
adenohyoyseaI reIease of TSH, lhe secrelion allern of TSH vas anaIyzed in
beagIe bilches, before and afler lhe induclion of rimary hyolhyroidism (Chaptcr
4).

Chaler 1
_________________________________________________________________________________
12

Some of lhe hysicaI changes observed in hyolhyroid dogs mimic lhose
of acromegaIy. Ior lhis reason, lhe uIsaliIe allern of grovlh hormone (GH)
secrelion vas invesligaled in seven eulhyroid beagIe bilches and 1.5 years afler lhe
induclion of rimary hyolhyroidism (Chaptcr 5).
The resuIls of lhe sludy described in Chaptcr 5 indicale lhal rimary
hyolhyroidism in dogs is associaled vilh increased GH reIease. In search for an
exIanalion for lhis, lhe effecl of inlravenous adminislralion of TRH on GH reIease
vas invesligaled in dogs vilh rimary hyolhyroidism and in heaIlhy conlroI
dogs (Chaptcr 6).
To lesl lhe main hyolhesis of lhis lhesis, seven beagIe dogs vere foIIoved
u for 3 years afler lhe induclion of rimary hyolhyroidism. Three of lhese dogs
vere foIIoved u for anolher 1.5 years vhiIe receiving I-lhyroxine.
AdenohyohyseaI funclion vas invesligaled al 2-monlh inlervaIs vilh lhe
combined inlravenous in|eclion of four hyolhaIamic reIeasing hormones (CRH,
GHRH, GnRH, and TRH) and measuremenl of lhe Iasma concenlralions of lhe
iluilary hormones ACTH, GH, LH, IRL, and TSH. In addilion, every 6 monlhs
lhe iluilary gIand vas visuaIized by comuled lomograhy. HislomorhoIogicaI
and immunohislochemicaI sludies of lhe iluilary gIands vere erformed in bolh
dogs vilh unlrealed hyolhyroidism and hyolhyroid dogs on subslilulion
lheray vilh I-lhyroxine (Chaptcr 7).
The abrulIy induced form of hyolhyroidism in lhe beagIe dogs
described in Chaptcr 7 may nol be reresenlalive of lhe graduaIIy deveIoing
sonlaneous disease in dogs of olher breeds, ages, and gender. Therefore, lhe same
adenohyohyseaI funclion lesls as described in Chaptcr 7 vere erformed in 14
dogs vilh sonlaneous rimary hyolhyroidism. The resuIls vere comared vilh
lhose of 13 dogs vilh a Iov Iasma T4

concenlralion, bul no lhyroid Iesion, i.e.,
dogs vilh non-lhyroidaI iIIness (Chaptcr 8).
In Chaptcr 9 lhe resuIls of lhe sludies are summarized and discussed.













Chaptcr 2

Gcncra! Intrnductinn


Chaler 2
_________________________________________________________________________________

14

GeneraI Inlroduclion
_________________________________________________________________________________


15
1. Anatnmy nI thc thyrnid

In dogs lhe lhyroid Iobes are silualed on lhe IaleraI surfaces of lhe lrachea,
from aboul lhe lhird lo lhe eighlh lracheaI ring. The righl Iobe is sIighlIy craniaI lo
lhe Iefl and nearIy louches lhe caudaI border of lhe Iarynx. They are covered
venlraIIy by lhe slernohyoid and slernolhyroid muscIes. In an aduIl dog veighing
aboul len kiIogram, each Iobe is aboul lvo cm by one cm by one-haIf cm, and lhe
combined veighl of lhe lvo Iobes is aboul one gram. ecause lhe Iobes are
reIaliveIy smaII and Iocaled undernealh lhe slernocehaIicus muscIe, lhey are
normaIIy nol aIabIe in lhe dog. The ma|or bIood suIy is via lhe craniaI lhyroid
arlery, a branch of lhe common carolid, and lhe rinciaI venous drainage is via
lhe caudaI lhyroid vein, vhich enlers lhe inlernaI |uguIar vein.
1

The basic slruclure of lhe lhyroid is unique for endocrine gIands,
consisling of foIIicIes of varying size lhal conlain coIIoid roduced by foIIicuIar
ceIIs. These cIoseIy acked shericaI unils, i.e. lhe foIIicIes, are invesled vilh a rich
caiIIary nelvork.
2
Vieved in hisloIogicaI seclions, lhe foIIicIes range from aboul
20 lo 250 m in diameler. The vaII of lhe foIIicIe is a singIe Iayer of lhyroid
eilheIiaI ceIIs. The foIIicuIar ceIIs are 4 lo 10 m in heighl,
3
and vary in heighl vilh
lhe degree of gIanduIar slimuIalion, becoming coIumnar vhen aclive and cubicaI
vhen inaclive. Their secrelory oIarily is direcled lovard lhe Iumen of lhe
foIIicIes.
2
The Iumen is fiIIed vilh a roleinaceous coIIoid lhal conlains a Iarge
gIycorolein caIIed lhyrogIobuIin, vilhin lhe sequence of vhich lhe lhyroid
hormones are synlhesized and slored. In random cIusler of foIIicIes, lhe erihery
of lhe coIIoid is vacuoIaled.
3

Accessory lhyroid lissue is quile common in lhe dog and may be Iocaled
anyvhere from lhe Iarynx lo lhe diahragm.
2,3
Il arises from lhyroid lissue
remaining al lhe IeveI of lhe lhyrogIossaI ducl and/or descending vilh lhe hearl
during embryonic deveIomenl.
4
Allemls lo induce hyolhyroidism in lhe dog
by surgicaI lhyroideclomy are nol consislenlIy successfuI because lhe accessory
lhyroid noduIes (s 2 mm) can undergo sufficienl hyerIasia lo suslain adequale
hormone roduclion.
2,3,5


2. Thyrnid physin!ngy

The funclion of lhe lhyroid gIand is lo generale lhyroid hormone in
quanlilies necessary lo meel lhe demands of eriheraI lissues. The lhyroids
secrele lhe hormones I-lhyroxine, or 3,5,3',5'-L-lelraiodolhyronine (T4), and in
much smaIIer quanlilies (aboul 20% of lhal of T4) lriiodolhyronine (T3). Mosl
circuIaling T3, lhe melaboIicaIIy aclive lhyroid hormone, is roduced in eriheraI
lissues by ouler ring deiodinalion of T4.
Chaler 2
_________________________________________________________________________________

16

2.1 lodlne metuboIlsm
The formalion of normaI quanlilies of lhyroid hormones deends on lhe
avaiIabiIily of adequale quanlilies of exogenous iodine lo aIIov lhyroidaI ulake.
The daiIy iodine requiremenl of an aduIl dog veighing 10-15 kg is aboul 140 g.
6

A! |i|iiun consumlion of mosl commerciaIIy manufaclured dry dog foods
rovides lhe average dog vilh a daiIy iodine inlake of al Ieasl 500 g, and some
foods rovide considerabIy more. Mosl iodine in lhe diel is reduced lo iodide in
lhe gaslroinleslinaI lracl. The absorlion of iodide begins immedialeIy and is
essenliaIIy comIele vilhin lvo hours. In lhe dog, iodide is aIso cIeared from lhe
Iasma by lhe arolid saIivary gIands and lhe gaslric mucosa. A smaII amounl of
iodide is normaIIy Iosl in lhe feces (aboul 20 lo 25 g er day al lhe usuaI IeveIs of
iodine inlake), ossibIy via secrelion in lhe coIon.
3


2.2 Throld hormones snthesls und secretlon
The biosynlhesis of lhyroid hormones is aIso unique among endocrine
gIands because lhe finaI assembIy of lhe hormones occurs exlraceIIuIarIy vilhin
lhe foIIicuIar Iumen. The foIIicuIar ceIIs of lhe lhyroid gIand concenlrale lhe
required amounls of iodide (lhe main buiIding bIock of lhyroid hormones) by
means a mechanism caIIed iodide lraing, vhich is accomIished by a
membrane rolein, lhe sodium-iodide symorler (NIS).
7,8,9
The lransorl of iodide
from lhe inlerfoIIicuIar caiIIaries inlo lhe lhyroid foIIicuIar ceIIs is an aclive
rocess via lhe ATI-deendenl NIS al lhe basiIar asecl of foIIicuIar ceIIs, and
resuIls in a lhyroid/Iasma concenlralion ralio of aboul 25.
2
The rocess deends
on lhe resence of a sodium gradienl across lhe basaI membrane of lhe lhyroid
ceII.
9
The foIIicuIar ceIIs of lhe lhyroid gIand concenlrale iodide, vhich diffuses
dovn a gradienl inlo lhe coIIoid. Tissues olher lhan lhe lhyroid, such as gaslric
mucosa, saIivary gIands, and choroid Iexus, aIso have an aclive lransorl
mechanism for iodide, aIbeil al a much Iover IeveI lhan lhe lhyroid. In conlrasl lo
lhe lhyroid gIands, lhese lissues do nol have lhe caacily for organic binding of
iodide. OnIy lhyroid foIIicuIar ceIIs accumuIale iodide in a TSH-deendenl
manner.
2

The assembIy of lhyroid hormones vilhin lhe foIIicuIar Iumen is
deendenl on lhe resence of a unique rolein, lhyrogIobuIin. This high-
moIecuIar-veighl gIycorolein is synlhesized in successive subunils on lhe rough
endoIasmic relicuIum, ackaged in lhe GoIgi aaralus vilhin lhe lhyroid
foIIicuIar ceIIs, and secreled inlo lhe coIIoid. Afler lhyrogIobuIin mRNA is
lransIaled in lhe rough endoIasmic relicuIum, lhe rolein is gIycosyIaled during
lransorl lhrough lhe GoIgi aaralus, vhere lhyrogIobuIin dimers are
incororaled inlo exocylic vesicIes. These vesicIes lhen fuse vilh lhe aicaI
basemenl membrane of foIIicuIar ceIIs, from vhere lhey are reIeased inlo lhe
GeneraI Inlroduclion
_________________________________________________________________________________


17
foIIicuIar Iumen. There, al lhe aicaI-coIIoid border, lhyrosine residues in
lhyrogIobuIin are iodinaled and aIso couIing lakes Iace.
10

Once in lhe lhyroid ceII, iodide arliciales in a series of reaclions lhal Iead
lo lhe synlhesis of lhe aclive lhyroid hormones. Iodide is raidIy oxidized by
lhyroid eroxidase (TIO) in lhe resence of H2O2 inlo a reaclive inlermediale lhal
is lhen incororaled inlo lyrosine residues vilhin lhe elide chain of
lhyrogIobuIin. This rocess, lermed organificalion, resuIls in lhe iodinaled lyrosine
residues monoiodolyrosine (MIT) and diiodolyrosine (DIT) vilhin lhe
lhyrogIobuIin moIecuIe. MIT and DIT, vhich are hormonaIIy inaclive, are
recursors of lhe hormonaIIy aclive iodolhyronines.
9
MIT and DIT lhen undergo
oxidalive couIing lo form lhe iodolhyronines, lhyroxine (T4), lriiodolhyronine
(T3), and reverse lriiodolhyronine (rT3), vhich remain bound lo lhyrogIobuIin unliI
secreled. Synlhesis of T4 from DIT requires lhe TIO-calaIyzed funclion of lvo DIT
moIecuIes lo yieId a slruclure vilh lvo diiodinaled rings Iinked by a bridge (lhe
couIing reaclion).
9
In a normaI dog vilh lhyroid gIands veighing one gram, lhe
lhyroid conlains aboul 900 g of T4, 75 g of T3, and 15 g of rT3.
3,11
Thiourea
comounds inhibil TIO, and lhereby bolh organic binding and couIing, vhich
makes lhese drugs suiled for lrealmenl of hyerlhyroidism.
Secrelion is inilialed by lhe ingeslion of lhyrogIobuIin inlo lhe foIIicuIar
ceII in lhe form of coIIoid droIels. Ingeslion may occur on lhe surface of lhe
foIIicuIar ceII by lhe formalion of seudoods, vhich encIose a smaII orlion of lhe
coIIoid, or by microinocylosis. Iseudood formalion is arlicuIarIy evidenl afler
slimuIalion of lhe lhyroid gIands vilh harmacoIogicaI doses of TSH, bul nol in
resling gIands, and il is lherefore IikeIy lhal microinocylosis is more imorlanl for
basaI secrelion. The coIIoid droIels acquire roleoIylic enzymes from Iysosomes,
robabIy al lhe lime of formalion,
12
and lhen lhey migrale lo lhe base of lhe ceII.
The iodolhyronines diffuse from lhe Iysosomes and lhen from lhe ceII inlo lhe
exlraceIIuIar sace, from vhere lhey enler lhe ad|acenl caiIIary.
UncouIed MIT and DIT in lhyrogIobuIin are aIso reIeased during
roleoIysis of lhe coIIoid droIels. In humans, MIT and DIT are very efficienlIy
deiodinaled vilhin lhe foIIicuIar ceII, and mosl of lhe iodide lhus generaled is
recycIed inlernaIIy lo be used again for iodinalion of lyrosoI grous of
lhyrogIobuIin. In lhe dog, mosl of lhe iodolyrosine iodine is reIeased back inlo lhe
circuIalion. This nonhormonaI iodine reIease by lhe canine lhyroid reresenls al
Ieasl lvo lhirds of lhe iodine iniliaIIy lraed and bound lo lyrosyI residues.
AIlhough onIy iodide has been idenlified during nonhormonaI iodine secrelion,
serum DIT IeveIs have been reorled lo be higher in lhe dog lhan in olher secies,
for examIe aboul four limes higher lhan in humans.
13
This reIalive high IeveI of
nonhormonaI iodine secrelion by lhe canine lhyroid is a ma|or faclor in lhe dog's

Chaler 2
_________________________________________________________________________________

18

reIaliveIy high iodine requiremenl, since iodine relurned lo lhe circuIalion is
sub|ecl lo renaI cIearance and fecaI excrelion.
The main secrelory hormonaI roducl of lhe lhyroid gIand is T4. The olher
lhyroid hormone, 3,5,3',-L-lriiodolhyronine (T3), is secreled in much smaIIer
quanlilies (aboul 20% of lhal of T4). In olher vords, lhe ralio of T4 lo T3 in lhe dog's
lhyroid secrelion is aboul 4:1.
14
The lolaI daiIy roduclion of lhe lvo hormones is
in a ralio of aboul 2:1 (aboul 30 g of T4 and 15 g of T3 in dogs veighing 10 lo 15
kg).
15
In dogs, aboul 40 lo 60 ercenl of lhe amounl of T3 roduced daiIy is
generaled via exlralhyroidaI conversion of T4 lo T3 in eriheraI lissues.
3
In
humans, aroximaleIy 80% of T3 is generaled oulside lhe lhyroid.
16,17,18,19
Dogs
roduce more lhan lvice lhe amounl of T4 and more lhan lhree limes lhe amounl
of T3 er kiIogram of body veighl lhan do humans.
3
Aboul 80% of lhe secreled T4
is deiodinaled lo form T3 (ouler ring deiodinalion of T4) and reverse T3 (rT3) (inner
ring deiodinalion of T4). Reverse T3 is lhe melaboIicaIIy inaclive 3,5',5'-
lriiodolhyronine.

2.3 Throld hormones trunsport und metuboIlsm
More lhan 99% of Iasma T4 and T3 is bound lo Iasma rolein, vilh T4
being more highIy bound lhan T3. Less lhan 0.05% of T4 and Iess lhan 0.5% of T3
circuIale free, in lhe unbound slale. The concenlralion of free lhyroid hormone,
vhich is avaiIabIe lo lissues and vhich is melaboIicaIIy aclive, delermines lhe
lhyroid slalus irreseclive of lhe Iasma lolaI lhyroid hormone concenlralion and
is mainlained conslanl by a feedback reguIalory syslem.
20
In lhe dog, lhe lhyroid-
binding roleins are lhyroid hormone-binding gIobuIin (TG), lranslhyrelin,
aIbumin, and aoIioroleins. Mosl T4 (aroximaleIy 60%) is bound lo TG, 17%
lo lhyroxine-binding reaIbumin (TIA), 12% lo aIbumin, and 11% lo an o1-
rolein.
21
In canine Iasma, T3 is bound lo aIbumin and one gIobuIin.
22
The IeveI of
circuIaling binding rolein may be changed by disease and harmacoIogicaI
agenls, and lhis may have considerabIe effecl on lhe lolaI Iasma lhyroid hormone
concenlralion, aIlhough lhe concenlralion of free lhyroid hormone may remain
unchanged. In dogs, lhyroid hormones bind vilh Iover affinily (veaker binding)
lo Iasma roleins lhan is lhe case in humans, and lhus, Iasma TT4 and TT3
concenlralions are Iover, free lhyroid hormone concenlralions are higher, and lhe
cIearance rales of lhese hormones are more raid lhan in humans. In arlicuIar,
dogs Iack a high-affinily binding gIobuIin comarabIe lo lhe inler-o lhyroxine-
binding gIobuIin (TG) of humans. TG in dogs is nol saluraled unliI lhe lolaI
lhyroxine concenlralion is aroximaleIy 6 limes lhe normaI vaIue.
19,23

In lhe dog, aboul 40% exlralhyroidaI T4 is resenl in lhe Iasma and mosl
of lhe remainder is laken u by lhe Iiver. The lolaI Iasma-equivaIenl sace of
dislribulion for T4 is aboul 12% of body veighl. The ralio T4 lo T3 in canine Iasma
GeneraI Inlroduclion
_________________________________________________________________________________


19
is aboul 25:1. The equivaIenl of 115% of exlralhyroidaI T4 and 205% of
exlralhyroidaI T3 is melaboIized and reIaced each day. Aboul 45% of T4 lurnover
is via deiodinalion and 55% is via fecaI excrelion, lhe biIiary cIearance of T4 is very
much higher in dogs lhan in humans. Aboul 70% of T3 lurnover is via deiodinalion
and 30% is via fecaI excrelion. olh lhe overaII rale of lurnover and lhe Ioss of lhe
hormone in feces are much higher in dogs lhan in humans.
3

OnIy aboul 15% of T4 or T3 is absorbed from lhe inlesline in dogs, vhereas
70% of T4 and essenliaIIy 100% of T3 is absorbed via lhis roule in humans. One
imorlanl resuIl of lhese differences is lhal lhe reIacemenl dose of lhyroid
hormone for lrealing hyolhyroidism in a 10-kg dog is equivaIenl lo lhal used in
an aduIl man.
3

Deiodinalion is lhe main alhvay of lhyroid hormone melaboIism.
24
The
deiodinalion of T4 occurs in eriheraI lissues, mainIy lhe Iiver and kidney. The
reIalive rales of ouler- and inner-ring monodeiodinalion of T4 delermine lhe
quanlily of melaboIicaIIy aclive hormone avaiIabIe. T4 is aclivaled by ouler ring
deiodinalion lo T3, vhereas bolh T4 and T3 are inaclivaled by inner ring
deiodinalion lo 3,3',5-lriiodolhyronine and 3,3'-diiodolhyronine, resecliveIy.
These reaclions are calaIyzed by lhree iodolhyronine deiodinases.
24,25
Many sludies
have reveaIed lhal lhere are recirocaI changes in T3 and rT3 IeveIs in humans. In a
number of non-lhyroidaI iIInesses and during fasling, fever, and olher silualions in
vhich il vouId aear lo be advanlageous lo Iimil calaboIism, lhe aclivalion
alhvay lo T3 is decreased and lhe inaclivaling alhvay lo rT3 is increased. The
changes in T3 and rT3 are reversed during convaIescence. SimiIar changes are
execled lo occur in lhe dog.

2.4 MetuboIlc uctlon of throld hormones
OnIy free hormone enlers ceIIs lo roduce a bioIogicaI effecl or a negalive
feedback effecl on lhe iluilary and hyolhaIamus. Thyroid hormone ulake is
faciIilaled by differenl lyes of lhyroid hormone lransorlers.
26,27,28
T3 enlers ceIIs
more raidIy (in arl because T3 is Iess firmIy bound lo Iasma roleins) and has a
more raid onsel of aclion lhan T4. Since T3 has aroximaleIy lhree lo five limes
lhe melaboIic olency of T4, aImosl aII lhe melaboIic aclion of lhyroid hormone can
be ascribed lo lhe aclion of T3. The nolion lhal lhe lhyroid ilseIf conlribules IillIe lo
lhe T3 ooI does nol aIy lo slales of lhyroid hyerfunclion, in vhich lhe T3/T4
ralio of lhe secrelory roducl increases. AIlhough T4 aIso has inlrinsic melaboIic
aclivily, il has been caIIed a rohormone vhose aclivalion by deiodinalion lo T3 is
a sle reguIaled individuaIIy by eriheraI lissue.
19

T3 binds lo secific nucIear binding siles, and lhis inleraclion may Iead lo
augmenled (or suressed) lranscrilion of DNA or lo moduIalion of RNA, and
lherefore lo a change in rolein synlhesis and enzyme aclion.
20
Thyroid hormone
Chaler 2
_________________________________________________________________________________

20

has a vide variely of hysioIogicaI effecls, vhich incIude lhe romolion of grovlh
and maluralion of lissues, deveIomenl of lhe neuroIogicaI and skeIelaI syslems,
reguIalion of lhe roduclion and lurnover of many subslances, and reguIalion of
many enzymalic reaclions. Thyroid hormones increase lhe melaboIic rale and
oxygen consumlion of mosl lissues (vilh lhe excelion of lhe aduIl brain, lesles,
ulerus, Iymh nodes, sIeen, and anlerior iluilary), have osilive inolroic and
chronolroic effecls on lhe hearl, increase lhe number and affinily of bela-
adrenergic recelors, enhance lhe resonse lo calechoIamines, and increase lhe
roorlion of aIha-myosin heavy chains. Thyroid hormones have calaboIic effecls
on muscIe and adiose lissue, slimuIale erylhrooiesis, and reguIale bolh
choIesleroI synlhesis and degradalion.
20,29


2.5 ReguIutlon of throld functlon
The lhyroid gIands arliciale vilh lhe hyolhaIamus and iluilary in a
cIassicaI negalive feedback conlroI Ioo. The hyolhaIamic-iluilary feedback
mechanism senses varialions in lhe avaiIabiIily of free lhyroid hormones and acls
lo correcl lhem. Thyrolroin or lhyroid-slimuIaling hormone (TSH) is lhe ma|or
reguIalor of lhe morhoIogicaI and funclionaI slales of lhe lhyroid, and is a 28-kD
gIycorolein secreled by lhe anlerior Iobe of lhe iluilary.
9,30
The TSH moIecuIe
consisls of an o- and a -subunil. The o-subunil is idenlicaI lo lhe o-subunil of
gonadolroins, vhereas lhe -subunil is dislincl and confers on lhe TSH moIecuIe
ils bioIogicaI aclivily. In bolh lhe dog and lhe cal lhe genes encoding lhe -
subunils of TSH have been cIoned.
31,32
TSH romoles lhyroid hyerlrohy and
hyerIasia, and slimuIales lhe synlhesis and secrelion of lhyroid hormones (T4
and Iesser amounls of T3) from lhe lhyroid gIand.
9
TSH secrelion is inhibiled
rimariIy by T3, vhich is roduced IocaIIy by 5'-deiodinalion (D2), and aIso by T3
derived from lhe syslemic ooI of free T3.
24
T3 acls direclIy on iluilary lhyrolroes
lo decrease TSH synlhesis and reIease, and is lhus lhe main delerminanl of Iasma
TSH concenlralions.
The selling of lhe T4/T3-TSH feedback Ioo is moduIaled by a
hyolhaIamic lrielide, TSH-reIeasing hormone (TRH), vhich slimuIales TSH
synlhesis and reIease. TRH is roduced by neurons in lhe lhyrolroic area of lhe
hyolhaIamus, as veII as in olher arls of lhe brain. TRH is reIeased from neurons
in lhe median eminence and is lransorled lo lhe iluilary via lhe orlaI
circuIalion. Il lhen allaches lo recelors on lhe iluilary lhyrolroes.
3,33,34

TSH binds lo (G-rolein-Iinked) recelors on lhe foIIicuIar ceII, aclivales
adenyIale cycIase and lhus enhances cycIic adenosine monohoshale (cAMI)
roduclion, lhereby increasing lhe rale of biochemicaI reaclions concerned vilh lhe
biosynlhesis and secrelion of lhyroid hormones.
2,35
Wilhin minules of TSH
adminislralion, lhere is an increase in lhe binding of iodide lo rolein,
GeneraI Inlroduclion
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21
iodolhyronine synlhesis, lhyrogIobuIin secrelion inlo lhe foIIicuIar Iumen, and
ingeslion of coIIoid via seudood formalion. Increased lraing of iodide foIIovs
afler an inlervaI of a fev hours. TSH remains allached lo ils recelors for a Iong
lime and secrelion conlinues for some lime even if TSH is removed. In lhe dog,
inlravenous adminislralion of TSH causes an increase in lhe reIease of iodide from
lhe lhyroid, foIIoved by increased secrelion of lhyroid hormones vilhin 15 lo 30
minules. When a reIaliveIy Iarge dose of 10 IU of TSH is given, Iasma T4
concenlralions conlinue lo rise for 8 lo 12 hours.
3

The hisloIogicaI aearance of lhe lhyroid is infIuenced by lhe IeveI of
circuIaling TSH. If lhe secrelion of TSH is suslained (hours or days), lhyroid
foIIicuIar ceIIs become more coIumnar and lheir Iumen becomes smaIIer.
36,37,38
The
oosile occurs in foIIicuIar ceIIs in resonse lo a decrease in circuIaling iluilary
TSH. In lhis silualion, lhyroid foIIicIes become grealIy enIarged and dislended
vilh denseIy slained coIIoid as a resuIl of decreased TSH-medialed endocylosis of
lhe coIIoid. The foIIicuIar ceIIs Iining lhe invoIuled foIIicIes are cubicaI and lhere
are fev endocylolic vacuoIes al lhe inlerface belveen lhe coIIoid and foIIicuIar
ceIIs. The IuminaI surface of lhe foIIicuIar ceIIs is fIallened.
2

InlralhyroidaI reguIalion of lhyroid funclion aIso occurs, vilh lhere being
an inverse reIalionshi belveen lhe conlenl of organic iodine in lhe gIand and lhe
rale of hormone formalion. This auloreguIalory mechanism slabiIizes lhe rale of
hormone synlhesis desile fIuclualions in lhe avaiIabiIily of iodine, and lends lo
mainlain a conslanl lhyroid hormone ooI.
9
This auloreguIalory mechanism vilhin
lhe lhyroid gIands is eseciaIIy imorlanl in silualions of insufficienl or excessive
iodine suIy, and enabIes an immediale adalalion lo acule iodide excess (e.g.,
disinfeclion of Iarge areas of skin vilh iodine) by bIocking organic binding,
rimariIy by Iovering lhe exression of lhe genes encoding TIO and NIS.
39
A
graduaI increase in iodine inlake (and hence in lhyroid iodide IeveIs) resuIls in a
graduaI increase in lhe amounl of iodide bound lo lhyrogIobuIin, bul beyond a
cerlain IeveI binding is rogressiveIy inhibiled. This mechanism is aIso knovn as
lhe WoIff-Chaikoff effecl.
40
The inhibilion of iodide binding is lransienl in normaI
animaIs and humans, and escae occurs afler severaI hours. On lhe olher hand, in
iodine deficiency lhyroid funclion is increased Iong before lhe lhyroid slores of
organic iodine (lhyrogIobuIin) are exhausled.

3. Hypnthyrnidism

Hyolhyroidism is lhe cIinicaI syndrome resuIling from a deficienl
roduclion of lhyroid hormones. Al aduIl age in aroximaleIy 95% of cases il is a
rimary lhyroid disorder. OnIy in 5% or Iess of cases is lhe disease of
(sura)iluilary origin.
19,41
Causes of secondary hyolhyroidism (deficiency of
Chaler 2
_________________________________________________________________________________

22

TSH) incIude iluilary maIformalion and iluilary neoIasia. Terliary
hyolhyroidism (deficiency of TRH) has yel lo be documenled unequivocaIIy in
lhe dog. The reorled revaIence of canine hyolhyroidism ranges from 0.2% lo
0.8%.
42,43


3.1 Acqulred prlmur hpothroldlsm
Il has been reorled lhal lhere are lvo dislincl and ossibIy unreIaled
forms of acquired rimary hyolhyroidism, each accounling for aboul haIf of lhe
cases of rimary hyolhyroidism in dogs:
20,44

1. Chronic Iymhocylic lhyroidilis: a rogressive auloimmune rocess
Ieads lo Iymhocylic infiIlralion and rogressive deslruclion of foIIicIes and
subsequenl disaearance of lhyroid lissue.
45
HisloIogicaI fealures are diffuse and
noduIar infiIlralion of lhe lhyroid gIand by Iymhocyles and Iasma ceIIs, invasion
of individuaI foIIicIes by Iymhocyles and macrohages, and neulrohiIs onIy
around necrolic eilheIium. In end-slage Iymhocylic lhyroidilis, lhe lhyroid has a
characlerislic increase in coIIagen surrounding foIIicIes, some degeneraling
foIIicuIar ceIIs, scallered infIammalory ceIIs, vilh a residuaI infiIlrale of
Iymhocyles and Iasma ceIIs, Iarge ackels of arafoIIicuIar ceIIs lhal congregale
because of lhe disaearance of lhe foIIicIes and coIIase of lhe slroma. The
immunoIogicaI and moIecuIar alhogenesis of auloimmune lhyroidilis in dogs has
nol yel been veII characlerized. A considerabIe amounl of lissue (al Ieasl 75% of
lhe foIIicIes) musl be deslroyed before lhe cIinicaI manifeslalions of lhyroid
deficiency become evidenl, and lhis may lake lvo lo lhree years.
2. Thyroid alrohy or lhe so-caIIed idioalhic form: in lhis form lhere is
lhyroid alrohy vilhoul infIammalory infiIlrale.
3,44,46
The hisloIogicaI changes
incIude lhe reIacemenl of lhe lhyroid arenchyma by adiose lissue vilh lhe
absence or minimaI resence of infIammalory ceII infiIlrale.
45
The alrohy is oflen
so comIele lhal hisloIogicaIIy lhere may be nolhing more lhan aralhyroid lissue,
adiose lissue, and lvo or lhree very smaII lhyroid foIIicIes in an enlire seclion.
There is aIso an increase in coIIagen around bIood vesseIs, and very smaII nesls of
Iymhocyles may be found ad|acenl lo ackels of arafoIIicuIar ceIIs.

Il has been queslioned vhelher idioalhic alrohy is reaIIy a searale
enlily or vhelher lhe absence of infIammalory ceIIs is lhe consequence of lhe
disaearance of foIIicuIar eilheIium and coIIoid and hence lhe anligenic slimuIus
for lheir conlinuing resence. Olher Iess common causes of acquired rimary
hyolhyroidism in dogs are deslruclion of lhe gIand by nonfunclionaI lhyroid
lumors and iodine deficiency.
20,46



GeneraI Inlroduclion
_________________________________________________________________________________


23
3.1.1. CIlnlcuI munlfestutlons
Irimary hyolhyroidism affecls mainIy young lo middIe-aged (1-6 years)
dogs. Il affecls maIes and femaIes equaIIy.
3,41,43
Thyroidilis usuaIIy remains
unnoliced, aIlhough lransienl signs of hyerlhyroidism (oIyuria/oIydisia) may
be iniliaIIy observed, robabIy due lo lhe reIease of lhyroid hormone inlo lhe
circuIalion during an aclive hase of deslruclive lhyroidilis. The deveIomenl of
rimary lhyroid hormone deficiency is sIov and insidious, vilh a graduaI onsel of
cIinicaI manifeslalions.
3,41
The iniliaI changes are neilher secific nor aIarming, and
il may be monlhs before lhe dog is resenled for velerinary examinalion. The
cIinicaI manifeslalions are reIaled lo a generaIized sIoving of lhe melaboIism, and
lhey invoIve nearIy aII organ syslems. Hovever, lhe symloms from a singIe organ
syslem may dominale.
3,41
Hyolhyroid dogs may have IillIe viIIingness lo exercise,
may be Iess enlhusiaslic during lheir usuaI exercise, and may sIee for Ionger
eriods during lhe day. Affecled dogs may have difficuIly in mainlaining normaI
body lemeralure and may seek varmlh, being reIuclanl lo go oulside in cooI
vealher. The somelimes observed generaIized Iocomolor robIems may be lhe
resuIl of lhe severe reduclion in Na
+
, K
+
-ATIase caacily in lhe skeIelaI muscIes.
47

Canine hyolhyroidism has aIso been reorled lo be associaled vilh generaIized
myoalhy, vilh increased Iasma concenlralions of crealinine hoshokinase.
48,49

Iven lhough obesily or veighl gain has been reorled in aboul 50% of
hyolhyroid dogs, overnulrilion is more common lhan hyolhyroidism in mosl
obese dogs.
20

There are differenl and variabIe dermaloIogicaI robIems: dry coal and
skin, seborrhea, excessive shedding, relarded grovlh of hair, Ioss of undercoal.
3,20,41

Hyolhyroidism is beIieved lo redisose lo recurrenl bacleriaI infeclions of lhe
skin, and yoderma has been reorled in 10% lo 23% of hyolhyroid dogs.
20,42,43

MaIassezia infeclions and demodicosis have been reorled in hyolhyroid dogs.
Irurilus may occur if concurrenl infeclion is resenl.
20
AIoecia vilh
hyerigmenlalion of lhe skin is oflen Iocaled on lhe venlraI and IaleraI surfaces of
lhe lrunk, caudaI surfaces of lhe lhighs, venlraI surfaces of lhe neck, lhe dorsaI
surface of lhe nose, lhe end of lhe laiI, lhe laiI gIand area, and lhe base of lhe ears.
Myxedema (culaneous mucinosis) is anolher dermaloIogicaI manifeslalion
of hyolhyroidism and is characlerized by lhickening of lhe skin due lo deosilion
of hyaIuronic acid in lhe dermis. Thyroid hormone deficiency decreases lhe
calaboIism of gIycosaminogIycans and hyaIuronic acid accumuIales in lhe dermis,
and, because il is hygroscoic, lhe dermis becomes edemalous.
50,51
This occurs
arlicuIarIy in lhe forehead and face, Ieading lo a uffy aearance and lhickened
foIds of skin above lhe eyes. This, logelher vilh sIighl bIeharolosis, gives some
hyolhyroid dogs a lragic exression.
3,41


Chaler 2
_________________________________________________________________________________

24

The uIse rale is oflen sIov, and bolh femoraI uIse and aex beal are
veak. The ICG may shov a Iov voIlage of lhe comIexes.
3,41,52
Hyolhyroidism
causes imaired myocardiaI conduclivily and funclion. These aIleralions in cardiac
funclion seem lo be of IillIe cIinicaI imorlance in dogs vilh olhervise normaI
cardiac funclion, bul may be reIevanl in dogs vilh underIying cardiac disease.
42

Hyolhyroidism has been reIaled lo severaI femaIe reroduclive
abnormaIilies, such as roIonged inlereslrous inlervaI, siIenl eslrus, ersislenl
aneslrus, sonlaneous aborlion, smaII or Iov-veighl Iillers, ulerine inerlia, and
veak and sliIIborn uies. Hovever, conlroversy exisls since evidence for lhese
associalions is veak, and in some earIy sludies in vhich hyolhyroidism vas
reorled lo cause reroduclive robIems, lhe diagnosis of hyolhyroidism vas
based soIeIy on lhe measuremenl of basaI Iasma lhyroid hormone concenlralions.
Moreover, anolher sludy faiIed lo find an associalion belveen reroduclive
robIems and hyolhyroidism.
53
Hyolhyroidism has aIso been reIaled lo differenl
maIe reroduclive robIems, such as Ioss of Iibido, leslicuIar alrohy,
hyosermia, and azoosermia. Hovever a roseclive sludy of six maIe beagIes
vilh hyolhyroidism induced vilh
131
I delecled none of lhese robIems over a
eriod of 2 years.
54

Some hyolhyroid dogs may suffer from eriheraI nerve dislurbances
vilh generaIized eriheraI neuroalhy.
20,55,56
VeslibuIar signs incIuding head liIl
and alaxia as veII as faciaI nerve araIysis have been associaled vilh
hyolhyroidism. In many cases, if lhe craniaI nerves (faciaI, lrigeminaI, and
veslibuIocochIear) are affecled, lhe signs are muIlifocaI and rogressive over
lime.
20
The alhogenesis of cenlraI veslibuIar dysfunclion in hyolhyroid dogs is
IikeIy muIlifacloriaI and may invoIve vascuIar encehaIoalhy, muIliIe funclionaI
melaboIic derangemenls of neuronaI or gIiaI ceII ouIalions secondary lo lhyroid
hormone deficiency, and dysfunclion or faiIure of IocaI lhyroid hormone lransorl
and melaboIic alhvays vilhin lhe brain.
57

Tvenly-five lo 40 ercenl of hyolhyroid dogs have miId normochromic,
normocylic, nonregeneralive anemia as lhe resuIl of decreased erylhrooielin
roduclion. Thyroid hormone deficiency may aIso direclIy reduce lhe roIiferalion
of bone marrov slem ceIIs.
29,58

Hyolhyroidism aIlers lhe moIecuIar and biochemicaI reguIalion of
carbohydrale, Iiid, and rolein melaboIism. olh hyerchoIesleroIemia and
hyerlrigIyceridemia resuIl from decreased melaboIism and cIearance of lhese
subslances from lhe bIood.
58


3.1.2. Dlugnosls
Thyroid gIand funclion can be examined by measuring Iasma
concenlralions of lolaI T4 (TT4), lolaI T3 (TT3), free T4 (fT4), free T3 (fT3), and canine
GeneraI Inlroduclion
_________________________________________________________________________________


25
TSH (cTSH), by erforming a TSH slimuIalion lesl, a TRH slimuIalion lesl, lhyroid
scinligrahy, a lhyroid biosy, and by delermining circuIaling anlilhyrogIobuIin
anlibodies (TgAb) and circuIaling anlibodies againsl lhyroid hormones (anli-T3
(AT3) and anli-T4 (AT4)).

3.1.2.1. TotuI T4 und totuI T3
The basaI Iasma TT4 concenlralion (lhe sum of rolein-bound and free
hormone circuIaling in bIood) is a more accurale indicalor of lhyroid funclion lhan
is Iasma TT3 because circuIaling T3 is rimariIy a roducl of lhe eriheraI
conversion of T4. IIasma TT4 (and TT3) concenlralions in dogs can be moderaleIy lo
severeIy deressed by non-lhyroidaI iIIness (NTI). IIIness in lhis conlexl comrises
virluaIIy aII non-lhyroidaI diseases, surgicaI and nonsurgicaI, lrauma, and
inadequale caIoric inlake.
59
NTI is aIso referred lo as eulhyroid sick syndrome
and may have muIliIe effecls on eriheraI T3 and T4 lransfer, dislribulion and
melaboIism: 1) decreased Iasma binding of T4 and T3 lo Iasma carrier roleins
(by disIacemenl of lhyroid hormones from Iasma-binding roleins, by inhibiling
lhe synlhesis of binding roleins, and by enhancing lhe cIearance of binding
roleins), 2) increased T4 degradalion, 3) decreased 5'monodeiodinase aclivily and
increased 5 monodeiodinase aclivily, resuIling in decreased T4 conversion lo T3
and increased conversion lo rT3, 4) increased melaboIic cIearance of T3, 5)
inhibilion of TSH secrelion, and 6) direcl inhibilion of lhe secrelion of hormones
by lhe lhyroid.
58,60

AImosl aII crilicaIIy iII dogs in an inlensive care unil have Iov TT3 and TT4
IeveIs.
61
Suslained moderale-lo-severe NTI resuIls in Iov Iasma TT4 and
uIlimaleIy fT4 IeveIs. These effecls of NTI on lhyroid hormone melaboIism are Iess
veII characlerized in dogs lhan in humans. In humans, lhese changes have been
considered as roleclive henomena associaled vilh lhe body's resonse lo iIIness
and nulrilionaI slale. A decrease in Iasma TT4 concenlralions in severaI diseases
in humans, dogs, and cals correIales vilh a higher morlaIily and lherefore may
have a rognoslic vaIue in severe disease.
19,62

Velerinary drugs vhich Iover lhyroid hormone concenlralions incIude
gIucocorlicoids, anliconvuIsanls, quinidine, nonsleroidaI anli-infIammalory drugs
(NSAIDs), suIfaanlimicrobiaIs, radioconlrasl agenls, and aneslhelics.
17,63-66
The
effecls of NSAIDs on lhyroid funclion are marked bul vary from drug lo drug.
AcelyIsaIicyIic acid can raidIy affecl lhyroid homeoslasis in dogs, exerling a more
ronounced effecl on TT4 concenlralions lhan on fT4 and/or TSH concenlralions,
vhich couId robabIy refIecl lhe disIacemenl of lhyroid hormones from serum
rolein binding siles.
66
HaII evaIualed lhe effecl of a slandard dosage of
lrimelhorim suIfamelhoxazoIe on lhyroid funclion lesls in dogs vilh yoderma
and normaI baseIine serum lhyroid hormone concenlralions.
67
The average serum
Chaler 2
_________________________________________________________________________________

26

TT4, bul nol TT3, concenlralion feII significanlIy during lhe lrealmenl eriod of 6
veeks. The TSH resonse aIso feII in severaI dogs, and radionucIide imaging
suggesled lhal lhe rearalion (IikeIy lhe suIfa comonenl) inlerfered vilh iodine
melaboIism by lhe lhyroid gIands.
ConsequenlIy, a Iov Iasma TT4 (and TT3) concenlralion may be due lo
rimary hyolhyroidism, secondary hyolhyroidism, or NTI. Iven lhough
hyolhyroidism can be ruIed oul if lhe TT4 concenlralion faIIs vilhin lhe reference
range for eulhyroid dogs,
19
aboul 10% of hyolhyroid dogs are missed vhen lhe
diagnosis reIies soIeIy on TT4 IeveIs, because a normaI Iasma TT4 concenlralion
can be observed in aroximaleIy 10% of hyolhyroid dogs, robabIy due lo lhe
resence of anli-T4 anlibodies (AT4).
44


3.1.2.2. lree T4 und free T3
The measuremenl of free or unbound T4 (fT4) is a more accurale indicalor
of lhe melaboIicaIIy aclive hormone lhan TT4.
19,60,68
When T4 binding caacily is
reduced, lhe circuIaling TT4 concenlralion is reduced bul lhe circuIaling fT4
concenlralion shouId be mainlained vilhin lhe normaI reference range.
TheorelicaIIy, free hormone concenlralions shouId nol be grealIy affecled by NTI
and consequenlIy shouId enabIe alienls vilh lhe eulhyroid sick syndrome lo be
dislinguished from alienls vilh lrue hyolhyroidism. Hovever, lhere can be
considerabIe overIa belveen fT4 vaIues in hyolhyroid and eulhyroid sick
alienls.
69
The fT4 concenlralions may aIso be adverseIy affecled in lhe eulhyroid
sick syndrome.
19,60,68-71
To comIicale inlerrelalion of fT4 measuremenls even more,
lhere is some overIa in fT4 vaIues in heaIlhy dogs and hyolhyroid dogs.
69

Given lhe exense of fT4 measuremenls and lhe simiIar overaII
erformance of lesl resuIls, TT4 measuremenls are currenlIy considered adequale
for evaIualing lhyroid funclion.
72
On lhe olher hand, fT4 measured by equiIibrium
diaIysis as a singIe lesl has been reorled lo be lhe lesl vilh lhe highesl
combinalion of sensilivily, secificily, and accuracy, and has lhe besl correIalion
vilh lhe cIinicaI slalus of lhe dog.
19
The fT4 equiIibrium diaIysis assay vas reorled
lo have a diagnoslic accuracy of 95% and a secificily of 93%, reresenling lhe
highesl vaIues for any singIe lhyroid funclion lesl.
70,72


3.1.2.3. TSH stlmuIutlon test
Ior severaI decades, lhe diagnosis of canine rimary hyolhyroidism vas
based on a Iov TT4 concenlralion lhal does nol resond afler 4 or 6 hours lo
slimuIalion vilh bovine TSH (bTSH).
73
This TSH slimuIalion lesl has aIso been
used as lhe goId slandard for dislinguishing dogs vilh hyolhyrodism from dogs
vilh NTI. If bolh basaI vaIue and vaIue afler TSH slimuIalion are exlremeIy Iov,
lhere is no doubl aboul lhe diagnosis of rimary hyolhyroidism. Hovever, il is
GeneraI Inlroduclion
_________________________________________________________________________________


27
knovn lhal in dogs vilh NTI, lhe resuIls of a TSH slimuIalion lesl may mimic
lhose of dogs vilh hyolhyroidism. The inlerrelalion is even more difficuIl vhen
lhe basaI vaIue is Iov and slimuIalion vilh TSH is foIIoved by a rise in lhe Iasma
T4 concenlralion. This may be observed in dogs vilh eulhyroid sick syndrome or
secondary hyolhyroidism. In dogs vilh secondary hyolhyroidism, lhe resonse
lo a singIe dose of TSH is quile variabIe and robabIy deends uon lhe degree
and duralion of subnormaI lhyroid aclivily. In mosl cases, a singIe dose of TSH
causes an increase in Iasma T4 concenlralion simiIar lo lhal seen in dogs vilh
drug-induced Iovering of Iasma T4. Hovever, in some cases lhe resonse lo a
singIe dose of TSH is quile smaII, and lherefore TSH slimuIalion (i.e. 10 IU daiIy,
subculaneousIy) is erformed on lhree conseculive days, lo ensure lhal Iasma T4
concenlralions are increased and simuIlaneousIy lo roduce normaI scinligrahic
images of lhe lhyroid gIand.
3
IinaIIy, in cases of arliaI hyolhyroidism, Iov basaI
Iasma T4 concenlralions lhal increase afler TSH adminislralion may aIso be
observed.
41,74
AIlhough lhe TSH slimuIalion lesl occasionaIIy caused serious
diagnoslic diIemmas, il has been regarded as lhe Ieasl inconcIusive biochemicaI
lhyroid funclion lesl for differenlialing belveen rimary hyolhyroidism and NTI
in dogs. The lesl assesses lhe funclionaI reserve of lhe lhyroid gIands, and il has
been slaled lhal aroximaleIy 75% of lhe lhyroid gIands musl be deslroyed before
lhyroid hormone roduclion is diminished.
19

The inlroduclion of a radioimmunoassay for hTSH made il ossibIe lo
measure T4 and TSH in a singIe Iasma samIe. The combinalion of a Iov T4 vaIue
and a high TSH vaIue suorls lhe diagnosis of rimary hyolhyroidism in
eoIe. As a resuIl of lhis deveIomenl in human endocrinoIogy, lhere vas no
Ionger a demand for bTSH for diagnoslic uroses, vhich became commerciaIIy
unavaiIabIe. Ior some lime, lhe unavaiIabiIily of bTSH Iimiled lhe use of lhe TSH
slimuIaling lesl, bul novadays recombinanl human TSH (rhTSH) can be used as
an aIlernalive.
75-78


3.1.2.4. cTSH
The deveIomenl of a canine secies-secific immunoassay for TSH
offered lhe ossibiIily of imroving our underslanding of lhe canine iluilary-
lhyroid axis.
79,80
The cTSH assay can be used as a screening lesl for
hyolhyroidism, comarabIe lo lhe vay lhe hTSH assay is used in human
medicine. As execled, in canine rimary hyolhyroidism lhere is oflen a
combinalion of a Iov Iasma T4 concenlralion and a high Iasma TSH
concenlralion vilhin lhe same Iasma samIe.
81
Hovever, severaI sludies have
reveaIed lhal aroximaleIy one fourlh lo one lhird of dogs vilh rimary
hyolhyroidism have a Iov Iasma T4 concenlralion and a Iasma TSH
concenlralion vilhin lhe reference range for eulhyroid dogs.
19,82
SeveraI
Chaler 2
_________________________________________________________________________________

28

exIanalions for lhis henomenon have been suggesled, such as concurrenl
secondary hyolhyroidism, faiIure of lhe TSH assay lo delecl aII isoforms of
circuIaling TSH, decrease in reviousIy high TSH vaIues in dogs vilh
hyolhyroidism vilh lime, and uIlradian fIuclualions in lhe circuIaling
concenlralion of TSH.
19
In humans, lhe serum TSH concenlralion can be lransienlIy
increased during recovery from severe iIIness.
51


3.1.2.5. TRH stlmuIutlon test
Measuremenl of lhe TSH resonse afler TRH adminislralion offers a raid,
safe, and effeclive melhod of invesligaling lhe reserve caacily of lhe iluilary
lhyrolroe ceII funclion. TRH is readiIy avaiIabIe and Iess exensive lhan rhTSH.
The TRH slimuIalion lesl is used in humans lo differenliale secondary
(hyohyseaI) from lerliary (hyolhaIamic) hyolhyroidism. TheorelicaIIy, lhe
TRH slimuIalion lesl can aIso be used in lhe diagnosis of rimary hyolhyroidism,
because exogenous TRH may acl as an amIifier lo exaggerale an abnormaIily in
TSH secrelion. Hovever, in dogs vilh rimary hyolhyroidism lhere is nol an
exaggeraled TSH resonse lo slimuIalion vilh TRH.
83,84


3.1.2.6. Throld sclntlgruph
Scinligrahic imaging of lhe lhyroid is exlremeIy usefuI in lhe diagnosis of
aII lhyroid disorders of dogs and cals.
3
Iodide, lhe main buiIding bIock of lhyroid
hormones, is acliveIy lransorled from lhe exlraceIIuIar fIuid inlo lhe lhyroid
foIIicuIar ceIIs. Tissues olher lhan lhe lhyroid, such as gaslric mucosa, saIivary
gIands, and choroids Iexus, aIso have an aclive lransorl mechanism for iodide.
In conlrasl lo lhe lhyroids, lhese lissues do nol have lhe caacily for organic
binding of iodide.
41
AII of lhese iodide-concenlraling lissues are aIso caabIe of
concenlraling olher slrucluraIIy reIaled monovaIenl anions, such as erlechnelale
(TcO4
-
). Ierlechnelale is nol organicaIIy bound in lhe lhyroid and hence ils
duralion vilhin lhe lhyroid is shorl. This roerly, logelher vilh ils shorl hysicaI
haIf-Iife, makes lhe radioaclive isoloe of erlechnelale (
99m
TcO4
-
) a vaIuabIe
radionucIide for imaging lhe lhyroid by scinliIIalion scanning.
41
The lechnique is
usefuI for differenlialing belveen hyolhyroidism and drug-induced Iovering of
Iasma T4 concenlralions.
3
CaIcuIalion of lhe ercenl ulake of
99m
TcO4
-
has roven
lo be very usefuI in lhe assessmenl of lhyroid funclion in humans and cals.
85


3.1.2.7. Throld uItrusonogruph
High-resoIulion uIlrasonic robes are novadays being aIied lo evaIuale
lhyroid voIume and echogenicily. Significanl differences have been found in
lhyroid voIume and mean cross-seclionaI surface area belveen eulhyroid and
hyolhyroid dogs, bul nol belveen heaIlhy eulhyroid dogs and dogs vilh
GeneraI Inlroduclion
_________________________________________________________________________________


29
eulhyroid sick syndrome. In eulhyroid dogs and eulhyroid sick dogs, lhe
arenchymaI echolexlure vas homogeneous and hyerechoic, vhereas lhe reIalive
lhyroid echogenicily of hyolhyroid dogs vas Iess. In arlicuIar, lhe lhyroid
voIume vas found lo be highIy secific rediclive of hyolhyroidism.
86,87


3.1.2.8. Throld blops
iosy of lhe lhyroid gIand vilh subsequenl hisloIogicaI evaIualion is lhe
mosl definilive lesl lo idenlify lhyroid alhoIogy.
58,60
Iine-needIe asiralion biosy
in non-neoIaslic lhyroid diseases is difficuIl because of lhe size of lhe gIand and
slruclures surrounding il.
60

An adequale samIe can be oblained by surgicaI excision of lhe caudaI
one-lhird of one lhyroid Iobe. Irimary hyolhyroidism is characlerized by alrohy
of foIIicuIar ceIIs and Ioss of coIIoid or infiIlralion of Iymhocylic aggregales vilh
foIIicuIar ceII deslruclion. Secondary hyolhyroidism is characlerized by fIallened
foIIicuIar ceIIs and lhe reIalive absence of coIIoidaI vacuoIes.
58


3.1.2.9. ClrcuIutlng untlthrogIobuIln untlbodles (TgAbs) und untlbodles ugulnst
throld hormone. untl-T3 (AT3) und untl-T4 (AT4)
ThyrogIobuIin is a foIIicuIar gIycorolein lhal conlains T3 and T4 and ils
rimary slruclure is highIy anligenic. When in lhe foIIicuIar coIIoid, lhyrogIobuIin
is reIaliveIy rolecled from immune surveiIIance, bul lhyroid damage or in|ury
oflen resuIls in lhe deveIomenl of TgAbs.
19,58
During lhe infIammalory rocess of
Iymhocylic lhyroidilis, anlibodies are reIeased inlo lhe circuIalion. In lhe dog,
lhese anlibodies are redominanlIy reaclive againsl lhyrogIobuIin. In humans, lhe
mosl common anligen lo vhich anlibodies are delecled in alienls vilh lhyroidilis
is TIO.
44
TIO, or lhyroid eroxidase, is lhe ma|or lhyroid microsomaI anligen.
Recombinanl human TIO is nov used for lhe deleclion of anlilhyroid microsomaI
anlibodies lhal are commonIy resenl in lhe serum of alienls vilh Hashimolo's
lhyroidilis.
51

Iulhyroid dogs may aIso have TgAbs. So, lhe resence of TgAbs is nol a
definilive indicalion lhal hyolhyroidism viII deveIo. Moreover, 40% of
hyolhyroid dogs do nol have deleclabIe TgAbs. The cIinicaI significance of a
osilive TgAbs liler musl be eslabIished. Hovever, because lhe incidence of such
anlibodies is significanlIy higher in hyolhyroid dogs lhan in eulhyroid dogs,
rouline measuremenl is IikeIy lo be of cIinicaI vaIue, arlicuIarIy for moniloring
dogs in earIy slages of lhe disease.
58
Of heaIlhy dogs vilh osilive TgAbs,
aroximaleIy 20% rogressed vilhin a year lo deveIo addilionaI cIinicaI or
Iaboralory abnormaIilies consislenl vilh hyolhyroidism. In heaIlhy animaIs vilh
osilive TgAbs (bul no olher cIinicaI or Iaboralory abnormaIilies), sequenliaI TSH
measuremenls shoved a highIy slalislicaIIy significanl increase in serum TSH
Chaler 2
_________________________________________________________________________________

30

concenlralion, even if lhe vaIues remained vilhin lhe normaI range. Dogs vilh
osilive TgAbs and normaI TT4, fT4, and TSH vaIues shouId be scheduIed for
foIIov-u moniloring, for examIe every 6 monlhs.
19

The lhyroid hormones T3 and T4 are simIe iodinaled amino acids of Iov
moIecuIar veighl. Wilh lhyrogIobuIin acling as halen, anlibodies againsl T3 and
T4 may be roduced during lhe deveIomenl of Iymhocylic lhyroidilis.
29
High
auloanlibody lilers may inlerfere vilh hormone immunoassay measuremenls. This
silualion shouId be susecled in a dog vilh an aarenlIy exlremeIy high vaIue of
T4 and/or T3 in lhe face of cIinicaI and hisloricaI evidence of hyolhyroidism.
19,44,58,88

Ior reasons lhal are nol cIear, lhe incidence of anli-T3 anlibodies aears lo be
grealer lhan lhal of anli-T4 anlibodies. The significance of anlilhyroid hormone
anlibodies is rimariIy in lheir indicalion of Iymhocylic lhyroidilis and in lhe
confusion lhey creale in lhe inlerrelalion of lhyroid funclion lesls.
58
Thyroid
hormone auloanlibodies may deveIo in cIinicaIIy eulhyroid dogs as veII as in
dogs vilh Iymhocylic lhyroidilis, and lherefore confirmalion of lhyroidilis in
lhese dogs is besl accomIished using a lhyroid biosy.
58

As in humans, lhe resence of T3 and T4 auloanlibodies aears lo be of no
cIinicaI significance lo lhe dog, since lhe binding caacily of lhe anlibody is easiIy
exceeded by lhe adminislralion of slandard dosages of I-lhyroxine.
58,88

Immunosuressive lheray does nol aear lo be necessary or varranled.
IvaIualion during lhyroid hormone reIacemenl lheray is rimariIy based on
cIinicaI imrovemenl (or a Iack lhereof). Theraeulic moniloring of lhe circuIaling
lhyroid hormone concenlralions is nol roulineIy recommended in lhese dogs
because auloanlibodies viII conlinue lo roduce surious resuIls.
58
In conlrasl, fT4
concenlralions measured by equiIibrium diaIysis do nol aear lo be affecled
adverseIy by lhe resence of auloanlibodies and may rovide vaIuabIe diagnoslic
informalion. Thus lhis assay is usefuI for lheraeulic moniloring if lhe dog has
auloanlibodies lo T4.
88


3.1.3. Dlugnostlc treutment ulth I-throxlne
Some aulhors roose a lheraeulic lriaI vilh Ievo-lhyroxine (I-lhyroxine)
lo diagnose hyolhyroidism. This aroach lakes lime lo see resuIls, and once
changes are observed inlerrelalion of lhe resuIls may be difficuIl. Time senl on a
lheraeulic lriaI may aIso aIIov an undiagnosed condilion lo rogress, and
lherefore every alleml shouId be made lo ruIe oul non-lhyroidaI iIIness. Once lhe
lheraeulic lriaI has been decided, an aroriale dosage of I-lhyroxine lvice daiIy
shouId be adminislered. An ob|eclive crilerion has lo be found lo |udge lhe success
of lhe lheray, as veII as lhe eriod of lime lo re-evaIuale lhe alienl. The
slrongesl evidence is gained by lhe observalion of cIinicaI imrovemenl and a cIear
resonse lo I-lhyroxine, foIIoved by a reIase vhen lhe medicalion is removed.
GeneraI Inlroduclion
_________________________________________________________________________________


31
Iurlhermore, lhe cIinician shouId be avare lhal lhere may be harmacoIogicaI
effecls of I-lhyroxine even in alienls vilhoul hyolhyroidism (i.e., oor coal
quaIily, seasonaI fIank aIoecia).
19,58,68
Thyroid hormones iniliaIIy slimuIale
anaboIic rocesses, such as hair grovlh, even in eulhyroid dogs.
89


3.1.4. Treutment
Thyroid hormone suIemenlalion is necessary lo resoIve lhe melaboIic
abnormaIilies associaled vilh hyolhyroidism, and lrealmenl has lo be given for
Iife. The goaI of lhyroid hormone reIacemenl lheray is lo aroximale lhe
secrelion of lhyroid hormone in a normaI dog.
68
AIlhough T3 is lhe melaboIicaIIy
aclive lhyroid hormone, il is nol lhe suIemenl of choice. Indeed, lhe necessily for
frequenl adminislralion (due lo ils shorl haIf-Iife, 4-6 g/kg body veighl every 8
hours), lhe inabiIily of lhe body lo moduIale lhe concenlralion lo vhich ceIIs are
exosed, and ils olenliaI for causing hyerlhyroidism make T3 unsuilabIe for
reIacemenl lheray. Sodium I-lhyroxine, a synlhelic roducl, is lhe lrealmenl of
choice in lhe dog.
29

The haIf-Iife of T4 in lhe dog has been eslimaled lo be belveen 10 and 16
hours. Dogs require higher lrealmenl doses and more frequenl adminislralion lhan
humans. OraI suIemenlalion vilh synlhelic I-lhyroxine can be slarled al a dose
of 10 g/kg lvice daiIy. Afler lvo monlhs, a conlroI examinalion is carried oul. The
Iasma T4 IeveI shouId be belveen 20 and 30 nmoI/I al 8-10 hours afler lhe Iasl
dose. If nol, lhe dose shouId be ad|usled. ecause of individuaI varialion in lhe
inleslinaI absorlion of T4, furlher ad|uslmenl may be needed.
41

A rimary advanlage of roviding lhe rohormone T4 is lhal lhe body is
given lhe oorlunily lo reguIale lhe amounl of T3 generaled by normaI
hysioIogicaI mechanisms. In facl, aroriale lheray resuIls in normaI IeveIs of
bolh T4 and T3 and a decrease in TSH lhrough lhe negalive feedback inhibilion of
lhe iluilary gIand. Thyroid hormones have mosl of lheir effecl lhrough aclivalion
of nucIear recelors lhal induce gene lransformalion and rolein roduclion.
Through lhis mechanism, lhyroid hormones have a duralion of aclion lhal exceeds
lheir resence in Iasma and makes once daiIy lrealmenl successfuI in mosl cases.
ecause of lhe considerabIe individuaI varialion in lhe harmacokinelics of lhyroid
hormones, some dogs do nol resond adequaleIy lo once-daiIy lrealmenl.
Therefore, iniliaI lvice-daiIy lrealmenl is rudenl lo ensure lhe highesl ossibiIily
of achieving a good lheraeulic resonse.
Trealmenl vilh lhyroid hormones is nol beneficiaI and is robabIy
delrimenlaI in dogs vilh NTI.
19,58
These dogs are robabIy nol hyolhyroid al lhe
ceIIuIar IeveI. In a sludy of dogs vilh hyercorlisoIism, a disease knovn lo be
associaled vilh Iov T4 concenlralions, lhyroidaI
131
I-ulake vas simiIar lo lhal in

Chaler 2
_________________________________________________________________________________

32

heaIlhy el dogs.
41
The delrimenlaI effecls of lrealmenl vilh lhyroid hormones in
lhese dogs are due lo lhe induclion of a slale of reIalive hyerlhyroidism in some
lissues and an increase in calaboIism. Il is inaroriale lo inslilule lhyroid
hormone lheray based on lhe soIe observalion of a Iov Iasma TT4 concenlralion
in an animaI vilhoul cIinicaI signs of hyolhyroidism. Thyroid hormone
adminislralion has been used in some silualions eilher inarorialeIy or vilh
IillIe scienlific evidence lo suorl ils use, examIes incIude lhe lrealmenl of
obesily, aIoecia, exercise inloIerance, and hyerIiidemia. Trealmenl vilh I-
lhyroxine shouId be Iimiled lo dogs vilh hyolhyroidism.
19


3.1.5. Prognosls
Irimary hyolhyroidism is one of lhe mosl gralifying diseases lo lreal,
because of lhe ease and comIeleness vilh vhich il resonds lo lrealmenl. Wilh
aroriale lrealmenl and foIIov-u examinalions (every haIf year), aII lhe
aIleralions associaled vilh hyolhyroidism are reversibIe, excel for some of lhe
neuroIogicaI changes. Therefore, lhe Iong-lerm rognosis is exceIIenl.
41

Resonse lo lrealmenl is usuaIIy noled vilhin one veek of inilialing
lrealmenl, vilh an increase in aIerlness and aclivily, and an imrovemenl in
allilude. Weighl Ioss can occur vilhin 2 lo 4 veeks and dermaloIogicaI
abnormaIilies begin lo resoIve afler 4 lo 6 veeks of lrealmenl. ComIele resoIulion
of aIoecia, hyerigmenlalion, and seborrhea may lake severaI monlhs. Hair Ioss
can be exacerbaled earIy in lrealmenl as hair foIIicIes enlering lhe anagen slage
shed lhe leIogen hairs.

4. Nnmcnc!aturc nI thc pituitary g!and

According lo lhe Nomina Analomica Velerinaria, lhe canine iluilary
gIand is comosed of lvo main arls 1) lhe adenohyohysis and 2) lhe
neurohyohysis.
1
The iluilary gIand is susended from lhe midIine of lhe
hyolhaIamus by a cyIindricaI slaIk. This slaIk is an exlension of lhe median
eminence of lhe hyolhaIamus and is caIIed lhe ars roximaIis neurohyohysis
(aIso caIIed lhe infundibuIum). The lhird venlricIe conlinues as an invaginalion
inlo lhe infundibuIum. The ars roximaIis neurohyohysis is conlinuous vilh
lhe dislaI enIargemenl, lhe ars dislaIis neurohyohysis, vhich is lhe ma|or
orlion of lhe neurohyohysis.
The adenohyohysis can be divided inlo lvo funclionaI unils 1) lhe
anlerior Iobe (ars infundibuIaris adenohyohysis and ars dislaIis
adenohyohysis) and 2) lhe ars inlermedia (ars inlermedia adenohyohysis).
In lhe dog, lhe Iargesl orlion of lhe anlerior Iobe Iies venlroroslraI lo lhe ars
dislaIis neurohyohysis, vhich is aImosl enlireIy surrounded by lhe anlerior
GeneraI Inlroduclion
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33
Iobe. The canine anlerior Iobe aIso exlends as a cuff or coIIar around lhe ars
roximaIis neurohyohysis and even enveIos arl of lhe median eminence. The
ars inlermedia is in direcl conlacl vilh lhe ars dislaIis neurohyohysis and is
searaled from lhe anlerior Iobe by lhe hyohyseaI cIefl or cavily, vhich is a
remnanl of lhe embryonic Ralhke's ouch.
The malure anlerior Iobe is ouIaled by al Ieasl five highIy differenlialed
lyes of endocrine ceIIs, vhich are cIassified according lo lhe lroic hormones lhey
roduce: somalolroic ceIIs secreling grovlh hormone (GH), Iaclolroic ceIIs
secreling roIacline (IRL), lhyrolroic ceIIs secreling lhyroid-slimuIaling
hormone, gonadolroic ceIIs secreling Iuleinizing hormone (LH) and foIIicIe-
slimuIaling hormone (ISH), and corlocolroic ceIIs synlhesizing lhe recursor
moIecuIe ro-oiomeIanocorlin (IOMC), vhich gives rise lo adenocorlicolroic
hormone (ACTH) and reIaled elides. Somalolroic ceIIs accounl for 50% or
more of lhe endocrine ceIIs in lhe anlerior Iobe, vilh lhe olher ceII lyes each
accounling for aboul 5-15% of lhe ceII ouIalion in lhe anlerior Iobe.
41,90


5. Functinna! aspccts nI thc pituitary g!and

5.1. HpothuIumlc-pltultur uxls
The hyolhaIamic-iluilary axis is lhe main axis of lhe neuroendocrine
syslem. The iluilary gIand is an essenliaI reguIalory inlerface, inlegraling signaIs
from lhe erihery and brain lo conlroI vilaI funclions (grovlh, reroduclion,
Iaclalion, basaI melaboIism, slress resonse).
91
HormonaI conlroI of iluilary gene
exression and ceII roIiferalion is inilialed during embryogenesis and conlinues
lhrough aduIlhood. Al aII slages, lhe ceIIs of lhe iluilary have a remarkabIe abiIily
lo roIiferale in resonse lo demand for a secific hormone. The hyolhaIamic-
iluilary axis consisls of lhree ma|or syslems: 1) a neuroendocrine syslem
connecled lo an endocrine syslem by a orlaI circuIalion, 2) a neurosecrelory
alhvay, and 3) a direcl neuraI reguIalion of endocrine secrelion.
The neuroendocrine syslem lhal is connecled lo lhe iluilary anlerior Iobe
invoIves cIuslers of elide- and monoamine-secreling ceIIs in lhe anlerior and
middIe orlions of lhe venlraI hyolhaIamus.
92
Their roducls, being reIeasing and
inhibiling faclors such as GH-reIeasing hormone (GHRH), somaloslalin,
lhyrolroin-reIeasing hormone (TRH), corlicolroin-reIeasing hormone (CRH),
and gonadolroin-reIeasing hormone (GnRH), are lransorled aIong lhe nerve
fibers lo lhe median eminence. Irom lhe median eminence lhey are reIeased inlo
lhe caiIIary vesseIs of lhe hyolhaIamic-iluilary orlaI syslem and lransorled
lo lhe iluilary, lo reguIale lhe secrelion of hormones from lhe anlerior Iobe.
Secificily is achieved by lhe resence of secific recelors on individuaI lyes of
iluilary anlerior Iobe ceIIs. Ior examIe, lhe inleraclion belveen hyolhaIamic
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GHRH and lhe GHRH recelor on lhe somalolroic ceIIs of lhe anlerior Iobe may
resuIl in GH secrelion. In addilion lo lhe hyolhaIamic hyohysiolroic
hormones, secrelion of anlerior Iobe hormones is reguIaled by feedback from
largel organs such as lhe lhyroids, adrenaIs, and gonads.
The neurosecrelory alhvay is invoIved in osmoreguIalion lhrough lhe
roduclion and reIease of vasoressin, and in arlurilion and nursing lhrough lhe
secrelion of oxylocin. The lvo neurohyohyseaI hormones are synlhesized by
ouIalions of magnoceIIuIar neurons, groued in lhe aravenlricuIar and
suraolic nucIei in lhe hyolhaIamus,
92
from vhich axons exlend lhrough lhe
iluilary slaIk and lerminale in lhe neurohyohysis on feneslraled bIood vesseIs.
Here, oxylocin and vasoressin are slored in secrelory granuIes vilhin lhe nerve
lerminaIs and are reIeased by exocylosis inlo lhe bIoodslream in resonse lo
aroriale slimuIi.
The iluilary ars inlermedia is oorIy vascuIarized and is direclIy
innervaled by redominanlIy doaminergic nerve fibers from lhe hyolhaIamus.
This direcl neuraI conlroI is IargeIy a lonic inhibilory infIuence. Desile high IeveIs
of bioaclive ACTH in lhe canine ars inlermedia,
93
lhe main hormone secreled by
lhe ars inlermedia is o-MSH.
94


5.1.1. Secretlon of ACTH
ACTH is synlhesized from a veII-characlerized recursor moIecuIe, ro-
oiomeIanocorlin (IOMC), vhich aIso gives rise lo a number of olher elides lhal
are co-reIeased vilh ACTH. ACTH secrelion by lhe anlerior Iobe is reguIaled by
lhe hyolhaIamus and cenlraI nervous syslem via neurolransmillers lhal cause lhe
reIease of hyohysiolroic hormones such as corlicolroin-reIeasing hormone
(CRH) and vasoressin. The Ialler are considered lo be lhe redominanl
slimuIaling neurohormones for ACTH in vivo.
95
ACTH aImosl excIusiveIy conlroIs
lhe synlhesis and reIease of gIucocorlicoids by lhe lvo inner zones of lhe adrenaI
corlex. In lurn, corlicosleroids inhibil ACTH reIease al muIliIe largel siles, of
vhich lvo have been idenlified unequivocaIIy: corlicolroic ceIIs in lhe anlerior
Iobe and neurons in lhe hyolhaIamus lhal roduce CRH and vasoressin. ACTH
and olher IOMC elides aIso exerl a grovlh-romoling effecl on lhe adrenaI
corlex.
96

The IOMC-roducing ceIIs of lhe ars inlermedia are resislanl lo
gIucocorlicoid suression, because lhey Iack gIucocorlicoid recelors.
Dexamelhasone relrealmenl reduces slimuIalion (vilh a doamine anlagonisl) of
ACTH reIease bul does nol affecl slimuIaled o-MSH reIease.
94
The inhibilory
hyolhaIamic doaminergic infIuence is heId resonsibIe for inhibiling lhe
exression of gIucocorlicoid recelors on ceIIs in lhe ars inlermedia, and
arlicuIarIy o-MSH-roducing ceIIs.
97,98

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5.1.2. Secretlon of GH
The hyolhaIamic neurohormones GH-reIeasing hormone (GHRH) and
somaloslalin Iay a cenlraI roIe in lhe secrelion of GH from lhe iluilary anlerior
Iobe.
99
Iisodic GH reIease can aIso be eIiciled by synlhelic GH-secrelagogues
(GHSs).
100
These GHSs eIicil lheir effecl on GH reIease by acling lhrough a secific
GHS recelor
101
lhal is nol aclivaled by GHRH or somaloslalin. In 1999, an
endogenous Iigand (ghreIin) for lhe GHS recelor vas idenlified.
102
In dogs,
eseciaIIy in young ones, ghreIin is a overfuI slimuIalor of GH reIease.
103
The
adrenergic syslem is lhe main neurolransmiller syslem lhal is invoIved in lhe
reguIalion of iluilary GH secrelion. GH secrelion is romoled by o2-adrenergic
agonisls such as cIonidine and xyIazine, vhereas -adrenergic agonisls are
inhibilory.
104

In lhe dog, excessive GH roduclion can be induced eilher by endogenous
rogeslerone or by exogenous rogeslagens used for eslrus revenlion.
105
The
rogeslagen-induced GH roduclion in lhis secies originales from foci of
hyerIaslic ducluIar eilheIium in lhe mammary gIand.
106
AIso in heaIlhy cycIic
bilches, lhe uIsaliIe allern of GH secrelion changes during rogression of lhe
IuleaI hase, vilh higher basaI GH secrelion and Iess GH secreled in uIses during
slages vilh a high Iasma rogeslerone concenlralion.
107
In Iine vilh lhis
observalion, ovarieclomy during lhe IuleaI hase resuIls is a Iover Iasma GH
concenlralion.
108

The effecls of GH can be divided inlo lvo main calegories: raid melaboIic
aclions and sIov (Iong-Iasling) hyerlrohic aclions. The acule melaboIic
resonses are due lo direcl inleraclion of GH vilh lhe largel ceII and resuIl in
enhanced IioIysis and reslricled gIucose lransorl across lhe ceII membrane due
lo anli-insuIin aclions. In addilion, GH has a direcl grovlh-romoling effecl by
slimuIaling ceII differenlialion. These sIov anaboIic effecls are mainIy medialed
via grovlh faclors lhal are synlhesized in lhe Iiver and are knovn as insuIin-Iike
grovlh faclors (IGIs). Their chemicaI slruclure has aroximaleIy 50% sequence
simiIarily vilh lhal of insuIin. In conlrasl lo insuIin, circuIaling IGIs are bound lo
binding roleins (IGIIs). As a resuIl of lhis binding lo carrier roleins, lhey have
a roIonged haIf-Iife, vhich is consislenl vilh lheir Iong-lerm grovlh-romoling
aclion. CircuIaling IGIs are imorlanl delerminanls in lhe reguIalion of body size,
by slimuIaling rolein synlhesis, chondrogenesis, and grovlh. IGI-1 exerls an
inhibilory effecl on GH reIease, mosl robabIy by slimuIaling lhe reIease of
somaloslalin and by an indirecl inhibilory infIuence al lhe iluilary IeveI.
109
In
addilion, GH ilseIf has a negalive feedback effecl al lhe hyolhaIamic IeveI.
110




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36

5.1.3. Secretlon of LH und lSH
UnIike lhe olher hormones of lhe iluilary anlerior Iobe, each of vhich is
secreled by a secific ceII, LH and ISH are secreled by lhe same lye of iluilary
ceII, lhe gonadolroe. The rimary slimuIalor of lhe synlhesis and uIsaliIe
secrelion of bolh LH and ISH is lhe decaelide gonadolroin-reIeasing hormone
(GnRH). Il mighl lherefore be execled lhal ISH and LH moIecuIes are discharged
concomilanlIy by gonadolroes. Hovever, in mosl mammaIian secies, incIuding
lhe dog,
111,112
differenliaI secrelion of LH and ISH has been demonslraled,
indicaling lhe resence of reguIalory mechanisms lhal aIIov indeendenl secrelion
of LH and ISH. One mechanism for lhe differenliaI conlroI of LH and ISH is
gonadaI feedback. Ior examIe, bolh eslradioI and inhibin can secificaIIy
suress ISH synlhesis and secrelion.
113
Olher imorlanl faclors in gonadolroin
conlroI are lhe frequency and amIilude of GnRH uIses, vhich have been shovn
lo differenliaIIy aIler LH and ISH gene exression as veII as lhe secrelion of lhese
hormones.
114
In addilion, a secific hyolhaIamic ISH-reIeasing decaelide has
been demonslraled in lhe ral.
115

Gonadolroins acl on lhe gonads lo slimuIale gonadaI hormone secrelion.
In lhe maIe animaI, LH slimuIales lhe synlhesis and reIease of lesloslerone from
lhe Leydig ceIIs of lhe leslis, vhereas ISH has a slimuIalory infIuence on lhe
secrelion of eslradioI and inhibin by lhe leslicuIar SerloIi ceIIs. In lhe femaIe
animaI, granuIosa, lheca, inlersliliaI, and IuleaI ceIIs are caabIe of secreling
hormones in resonse lo LH and ISH. The lye and amounl of hormones reIeased
vary according lo lhe morhoIogicaI slalus of lhe foIIicIe and corus Iuleum. The
allerns of secrelion of LH and ISH logelher vilh lhe morhoIogicaI and secrelory
changes in lhe ovaries delermine lhe secies-secific configuralion of lhe eslrous
cycIe.

5.1.4. Secretlon of PRL
The redominanl effecl of lhe hyolhaIamus on IRL secrelion by lhe
iluilary anlerior Iobe is inhibilory. The biogenic amine doamine is recognized as
lhe main inhibilory neuraI signaI in lhe reguIalion of IRL reIease.
116
eside
inhibilory doaminergic lone, severaI subslances are knovn lo have IRL-reIeasing
aclivily, such as serolonin, vasoaclive inleslinaI elide, and TRH.
104,117
A secific
IRL-reIease-romoling elide in lhe hyolhaIamus has been idenlified and
characlerized.
118,119

Neurogenic faclors aIso infIuence IRL secrelion. SuckIing and miIking are
aImosl immedialeIy foIIoved by IRL reIease. GonadaI sleroids are aIso imorlanl
moduIaling faclors in lhe conlroI of IRL secrelion. Islrogens raidIy induce and
enhance lhe IRL resonse lo TRH in dogs.
120
In addilion, rogeslerone may
moduIale IRL secrelion in lhe dog. IRL secrelion increases during lhe second arl
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37
of lhe IuleaI hase in nonregnanl bilches,
121
aIbeil Iess lhan in regnanl and
overlIy seudoregnanl bilches.
122,123


5.1.5. Secretlon of TSH
Iiluilary TSH secrelion is slimuIaled by hyolhaIamic TRH and inhibiled
by cenlraI doaminergic and somaloslalinergic mechanisms, as veII as by an
inhibilory aclion of lhyroid hormones al cenlraI, hyolhaIamic, and iluilary
IeveIs.
124
TSH secrelion is rimariIy inhibiled by 3,3',5 lriodolhyronine (T3), derived
from lhe ooI of circuIaling free T3 and roduced IocaIIy from lhyroxine (T4) by
lye II deiodinase.
24

A number of olher endogenous and exogenous agenls have been reorled
lo suress TSH secrelion, e.g. dobulamine, inlerIeukins, lumor necrosis faclor-2,
henyloin, doamine agonisls, somaloslalin anaIogues, serolonin, and
gIucocorlicoids.
9
CorlisoI and olher gIucocorlicoids aIso suress lhe TRH-induced
reIease of TSH. IroslagIandins and aIha-adrenergic agonisls increase TSH
secrelion.
Informalion concerning lhe dynamic secrelion of TSH in lhe dog is scarce.
The mean basaI TSH IeveIs are reIaliveIy Iov in eulhyroid dogs,
83
vhich couId be a
consequence of lhe reIaliveIy high iodine conlenl of induslriaI dog food.

6. Ana!ysis nI pu!sati!c sccrctinn nI adcnnhypnphysca! hnrmnncs

Il is nov generaIIy acceled lhal secrelion of aII adenohyohyseaI
hormones occurs in an uIlradian uIsaliIe fashion, and lhal lhis allern of secrelion
reresenls an imorlanl comonenl of neuroendocrine signaIing
mechanisms.
125,126,127
The uIlradian allerns of adenohyohyseaI hormones reIease
are driven by signaIs from lhe hyolhaIamus lo iluilary ceIIs, vhich are
moduIaled in lurn by inlrailuilary aracrine and aulocrine faclors, and aIso by
syslemalicaIIy derived feedback hormones.
The uIsaliIe secrelion allerns of adenohyohyseaI hormones can be
delermined by coIIecling bIood samIes seriaIIy over a eriod of hours or days.
The samIes are assayed and lhe resuIls are Iolled grahicaIIy lo enabIe
recognilion of lhe uIsaliIe secrelion allern. Differenl melhods can be used lo
anaIyze lhe dala. TradilionaIIy, grahs have been given lo a bIinded scorer vho
idenlifies lhe eaks by visuaI inseclion. Wilh lhis lechnique il is difficuIl lo slale
reciseIy lhe crileria used for uIse idenlificalion, vhich comIicales
communicalion among differenl cenlers sludying simiIar robIems, and oinls lo
lhe need for more ob|eclive melhods.
128
ConsequenlIy, a number of comuler
rograms have been deveIoed for comuler-assisled anaIysis of uIsaliIe
secrelion allerns. These comuler rograms have lo exlracl lhe reIevanl
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38

informalion from lhe secrelory allerns and dislinguish il from assay noise. A
melhod used quile oflen is deconvoIulion anaIysis.
129
DeconvoIulion lechniques
assume a one- or muIliIe-comarlmenl modeI and use lhe differenl comonenls
of lhe Iasma haIf-Iife of a secific adenohyohyseaI hormone lo eslimale lhe
secrelory aclivily of lhe iluilary gIand. Anolher, more descrilive, comuler
rogram lhal is used frequenlIy for invesligaling uIsaliIe secrelion allerns is lhe
IuIsar rogram. In conlrasl lo deconvoIulion lechniques lhal caIcuIale lhe acluaI
secrelion of lhe iluilary gIand, lhe IuIsar rogram onIy anaIyses lhe rhylhm in
Iasma hormone concenlralion, i.e., il rovides informalion aboul lhe hormonaI
miIieu lo vhich lhe largel gIand is exosed. ecause lhe IuIsar rogram does nol
reIy on knovIedge of eIiminalion kinelics, il can be used more easiIy vhen lhis
informalion is Iacking.
ecause lhe secrelory rofiIes of adenohyohyseaI hormones resenled
in lhis lhesis vere anaIyzed by means of lhe IuIsar rogram, some discussion of
lhe rogram is aroriale here. The IuIsar rogram, deveIoed by Merriam and
Walcher (1982),
128
idenlifies adenohyohyseaI hormone uIses based on lheir
amIilude and duralion vilh resecl lo a smoolhed baseIine (using a veighed
Iinear regression of hormone IeveI versus lime vilhin a cerlain lime vindov),
laking inlo accounl lhe slandard devialion of lhe assay al a given hormone
concenlralion. Iirsl, lhe rogram uses a robusl smoolhing lechnique lo generale a
smoolhed baseIine lhal omils eaks or lrends vilh lime conslanls Iess lhan 6-12 h.
The smoolhed baseIine is sublracled from lhe originaI dala and lhen lhe difference,
lhe residuaI series, is examined for lhe resence of eaks. The slandard devialion
of lhe assay is caIcuIaled al each oinl and lhe residuaIs are rescaIed in lerms of
lhis unil. Ieaks are idenlified as individuaI series of n hormone IeveIs above lhe
smoolhed baseIine, in vhich aII oinls musl have a magnilude of al Ieasl G(n). The
vaIues of G are cul-off crileria based on lhe vidlh of lhe eak. The narrover lhe
eak, lhe higher lhe residuaI hormone IeveI shouId be in order lo be idenlified as a
significanl eak. A oinl in lhe scaIed residuaI series is acceled as forming arl of
a eak if il aIone is higher lhan a cerlain cul-off vaIue G(1), or if il beIongs lo a air
of ad|acenl oinls bolh higher lhan a second Iover cul-off G(2), or a lriIel aII
higher lhan G(3), and so on, u lo G(5). Thus lhe aIgorilhm seIecls bolh narrov
high eaks and broader eaks lhal may be Iover. The user seIecls lhe G(n) for each
hormone based on lheorelicaI consideralions or a seIecl of caIibralion dala series.
In addilion, G(n) vaIues have been reorled lhal are chosen in such a vay lhal lhe
robabiIily of faIseIy cIaiming lhal lhere is a eak in a series vilhoul eaks is
beIov a given error rale, say 5%. To assure lhal lhe significanl eaks do nol
infIuence lhe caIcuIalion of lhe smoolhed baseIine, lhe smoolhing lechnique is
reealed, vhereby lhe residuaI hormone IeveIs of significanl eaks are assigned a
reduced veighl, and lhe subsequenl residuaI series is again examined for lhe
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39
resence of eaks. This rocess is ileraled unliI lhere are no furlher changes. Nexl,
each eak is examined once more lo delermine vhelher il can be resoIved inlo lvo
or more overIaing eaks.
128

Wilh regard lo lhe inlerrelalion of uIsaliIe secrelion allerns, some
melhodoIogicaI asecls shouId be laken inlo accounl. Iirsl, a high recision and
sensilivily of deleclion melhods are cruciaI for lhe anaIysis of uIsaliIe secrelion
allerns. A high inlra-assay coefficienl of varialion viII make il quile difficuIl lo
dislinguish significanl hormone uIses from assay noise. SecondIy, hormone-
secific resonses are delermined by lhe bioaclivily of lhe hormone. Hovever,
Iasma hormone concenlralions are measured by immunoassays ralher lhan
bioassays. ThirdIy, hormone secrelion allerns are measured as a reIaliveIy smaII
number of oinls, because on-Iine measuremenls of hormones are nol yel
avaiIabIe. The shorler lhe samIing inlervaI, lhe higher lhe chance lhal significanl
hormone uIses are indeed delecled. Therefore, lhe IuIsar rogram viII nol be
abIe lo exlracl lhe lrue underIying allern of informalion, bul lhis melhod
reresenls ob|eclive means for comarison of differenl dala sels.
126

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#

1. HuIIinger RL 1993 The endocrine syslem. In: MiIIer's Analomy of lhe Dog. 3
rd
edilion. Ivans
HI (Id). IhiIadeIhia, IA: W Saunders. :559-585.
2. Caen CC 2000 Analomy. In: Werner & Ingbar's The Thyroid. A IundamenlaI and CIinicaI
Texl. 8
lh
edilion. raverman LI and Uliger RD (Ids.). IhiIadeIhia, IA: Liincoll WiIIiams
and WiIkins. A WoIlers KIuver Comany. :20-43.
3. eIshav I 1983 Thyroid disease. In: Texlbook of Velerinary InlernaI Medicine, Diseases of
lhe Dog and lhe Cal. 2
nd
edilion. Illinger S} (Id.). IhiIadeIhia, IA: W Saunders. :1592-
1614.
4. Iessing MH, Zaborsky I 1966 |On lhe demonslralion of inlralhoracic lhyroid lissue in dogs
by means of I-131j. Irankf Z IalhoI 75:14-24.
5. Kameda Y 1972 The accessory lhyroid gIands of lhe dog around lhe inlraericardiaI aorla.
Arch HisloI }n 34:375-391.
6. eIshav I, Cooer TI, ecker DV 1975 The iodine requiremenl and infIuences of iodine
inlake on iodine melaboIism in lhe aduIl beagIe. IndocrinoIogy 96:1280-1291.
7. Dohan O, De Ia Vie|a A, Iaroder V, RiedeI C, Arlani M, Reed M, Ginler CS, Carrasco N 2003
The sodium/iodide symorler (NIS): characlerizalion, reguIalion, and medicaI significance.
Indocr Rev 24:48-77.
8. Krenning II, Wiersinga WM, GIinoer D 2007 SchiIdkIier In: IndocrinoIogie Krenning II,
Wiersinga WM, Lamberls SW}, NobeIs I, ouiIIon R (Ids.). 3
e
druk. Maarssen: IIsevier 139-
159.
9. Larsen IR, Davies TI, SchIumberger M-}, Hay ID 2008 Thyroid hysioIogy and diagnoslic
evaIualion of alienls vilh lhyroid disorders. In: WiIIiams Texlbook of IndocrinoIogy, 11
lh

edilion. Kronenberg HM, MeImed S, IoIonsky KS, Larsen IR, (Ids). IhiIadeIhia, IA: W
Saunders. :299-332.
10. Cooer DS, Greensan IS, Ladenson IW 2007 The Thyroid GIand. In: Greensan's asic &
CIinicaI IndocrinoIogy. 8
lh
edilion. Garderner DG and Shoback D (Ids.). Nev York: McGrav-
HiII. :209-280.
11. Laurberg I 1978 SeIeclive inhibilion of lhe secrelion of lriiodolhyronine from lhe erfused
canine lhyroid by royIlhiouracyI. IndocrinoIogy 103:900-905.
12. Nuez IA, eIshav I, Gershon MD 1972 A fine slrucluraI sludy of lhe highIy aclive lhyroid
foIIicuIar ceII of lhe African basen|i dog. Am } Anal 133:463-481.
13. NeIson }C, Weis RM, IaImer I}, Levis }I, WiIcox R 1975 Serum diiodolhyrosine. } CIin
IndocrinoI Melab 41:1118-1124.
14. Laurberg I 1976 T4 and T3 reIease from lhe erfused canine lhyroid isoIaled in silu. Acla
IndocrinoI 83:105-113.
15. eIshav I, arandes M, ecker DV, erman M 1974 A modeI of iodine kinelics in lhe dog.
IndocrinoIogy 95:1078-1093.
GeneraI Inlroduclion
_________________________________________________________________________________


41
16. Kalein IM, Hooes MT, Ierguson DC el aI. 1990 Comarison of reverse lriodolhyronine
dislribulion and melaboIism in normaI dogs and humans. IndocrinoIogy 126:2003-2014.
17. Kalein IM, Moore GI, Ierguson DC al aI. 1992 Iffecls of rednisoIon on lhyroxine and
3,5,3'-lriiodolhyronine melaboIism in normaI dogs. IndocrinoIogy 130: 1669-1679.
18. Kalein IM, Moore GI, Ierguson DC al aI. 1993 Thyroxine and lriiodolhyronine dislribulion
and melaboIism in lhyroxine-reIaced alhyreolic dogs and normaI humans Am } Ihys
(IndocrinoI Melab 27) 264: I90-102.
19. Ierguson DC 2007 Tesling for hyolhyroidism in dogs. Vel CIin Norlh Am SmaII Anim Iracl
37:647-669.
20. Scoll-Moncrieff }C 2007 CIinicaI signs and concurrenl diseases of hyolhyroidism in dogs and
cals. Vel CIin Norlh Am SmaII Anim Iracl 37:709-722.
21. Ierguson DC 1988 The effecl of nonlhyroidaI faclors on lhyroid funclion lesls un dogs.
Comendium SmaII AnimaIs Conlinuing Iducalion 10:1365-1376.
22. Refeloff S, Robin NI, Iang VS 1970 Iaramelers of lhyroid funclion in serum of 16 seIecled
verlebrale secies: A sludy of II, serum T4, free T4, and allern of T4 and T3 binding lo
serum roleins. IndocrinoIogy 86:793-805.
23. Ierguson DC 1984 Thyroid funclion lesls in lhe dog. Vel CIin Norlh Am SmaII Anim Iracl 14:
783-808.
24. Kuier GG}M, Kesler MHA, Ieelers RI, Visser T} 2005 iochemicaI mechanims of lhyroid
hormone deiodinalion. Thyroid 15:787-98.
25. ianco A and Kim W 2006 Deiodinases: imIicalions of lhe IocaI conlroI of lhyroid hormone
aclion. } CIin Invesl 116: 2571-2579.
26. Visser T} 2007 Thyroid hormone lransorles. Horm Res 68: 28-30.
27. Visser WI, Iriesema ICH, }ansen }, Visser T} 2008 Thyroid hormone lransorl in and oul of
ceIIs. Trends IndocrinoI Melab 2008 19:50-56.
28. Roberls LM, Woodford K, Zhou M, Iack DS, Haggerly }I, Tale IH, Grindslaff KK,
Mengesha W, Raman C, Zerangue N 2008 Ixression of lhe lhyroid hormone lransorles
MCT8 (SLC16A2) and OATI14 (SLC01C1) al lhe bIood-brain barrier. IndocrinoIogy 2008
|Iub ahead of rinlj.
29. Scoll-Moncrieff }CR and GuliII-Yoran L 2005. Hyolhyroidism. In: Texlbook of Velerinary
InlernaI Medicine. Diseases of lhe dog and lhe cal. 6
lh
edilion. Illinger S} and IeIdman IC
(Ids.). IhiIadeIhia, IA: W Saunders. :1535-1544.
30. Dremier S, MiIenkovic, Iancquaerl S, Dumonl }I, DoskeIand SO, Maenhaul C, Roger II
2007 CycIic adenosine 3',5'-Monohoshale (cAMI)-deendenl rolein kinases, bul nol
exchange roleins direclIy aclivaled by cAMI (Iac), mediale lhyrolroin/cAMI-deendenl
reguIalion lhyroid ceIIs. IndocrinoIogy 148:4612-4622.
31. Yang X, McGrav RA, Su X, Kalakam I, Grosse WM, Li OW, Ierguson DC 2000 Canine
lhyrolroin bela-subunil gene: cIoning and exression in Ischerichia coIi, generalion of
monocIonaI anlibodies, and lransienl exression in lhe Chinese hamsler ovary ceIIs. Domesl
Anim IndocrinoI.18:363-78.
Chaler 2
_________________________________________________________________________________

42

32. RayaIam S, Iizenslal LD, Hoenig M, Ierguson DC 2006 CIoning and sequencing of feIine
lhyrolroin (fTSH): helerodimeric and yoked conslrucls. Domesl Anim IndocrinoI 30:203-17.
33. ScanIon MI and Tofl AD 2000 ReguIalion of lhyrolroin secrelion. In: Werner & Ingbar's The
Thyroid. A IundamenlaI and CIinicaI Texl. 8
lh
edilion. raverman LI and Uliger RD (Ids.).
IhiIadeIhia, IA: Liincoll WiIIiams and WiIkins. A WoIlers KIuver Comany. :234-253.
34. Lov M} 2008 NeuroendocrinoIogy. In: WiIIiams Texlbook of IndocrinoIogy. 11
lh
edilion.
Kronenberg HM, MeImed S, IoIonsky KS, Larsen IR (Ids). IhiIadeIhia, IA: W Saunders,
: 85-154.
35. Wynford-Thomas D, Smilh I, WiIIiams ID 1987 IroIiferalive resonse lo cycIic AMI
eIevalion of lhyroid eilheIium in susension cuIlure. MoI CeII IndocrinoI 51:163-166.
36. Iricson LI, Ingslrm G 1978 Quanlilalive eIeclron microscoic sludies on exocylosis and
endocylosis in lhe lhyroid foIIicIe ceII IndocrinoIogy 103: 883-892.
37. CoIIins WT, Caen CC 1980 UIlraeslrucluraI and funclionaI aIleralions of lhe ral lhyroid
gIand roduced by oIychIorinaled bihenyIs comared vilh iodide excess and deficiency,
and lhyrolroin and lhyroxine adminislralion. Virchovs Arch 33: 213-231.
38. Many MC, Denef }I, Haumonl S, van den Hove-Vandenbroucke MI, Cornelle C, eckers C
1985 MorhoIogicaI and funclionaI changes during lhyroid hyerIasia and invoIulion in
C3Hmice: effecls of iodine and 3,5,38-lriiodolhyronine during invoIulion. IndocrinoIogy
116:798-806.
39. Uyllersrol N, IeIgrims N, Carrasco N, Gervy C, Maenhoul C, Dumonl }I, Miol I 1997
Moderale doses of iodide in vivo inhibil ceII roIiferalion and lhe exression of
lhyroeroxidase and Na
+
/I
-
symorler mRNAs in dog lhyroid. MoI CeII IndocrinoI 131:195-
203.
40. WoIff } and Chaikoff IL 1948 IIasma inorganic iodide as a homeoslalic reguIalor of lhyroid
funclion } ioI Chem 174: 555.
41. Ri|nberk A 1996 Thyroids. In: CIinicaI IndocrinoIogy of Dogs and Cals. Ri|nberk A (ed).
Dordrechl, KIuver Academic IubIishers, :35-39.
42. Ianciera DL 1994 Hyolhyroidism in dogs: 66 cases (1987-1992) } Am Vel Med Assoc 204:
761-767.
43. Dixon RM, Reid SW, Mooney CT 1999 IidemioIogicaI, cIinicaI, hemaloIogicaI and
biochemicaI characlerislics of canine hyolhyroidism. Vel Rec 145: 481-487.
44. Graham IA, RefsaI KR, Nachreiner RI 2007 IlioalhoIogic findings of canine
hyolhyroidism. Vel CIin Norlh Am SmaII Anim Iracl 37: 617-631.
45. GosseIin S}, Caen CC, Marlin SL 1981 HisloIogic and uIlraslrucluraI evaIualion of lhyroid
Iesions associaled vilh hyolhyroidism in dogs. Vel Ialh 18: 299-309.
46. Kemainen R} and CIark TI 1994 Ilioalhogenesis of canine hyolhyrodism. Vel CIin
Norlh Am SmaII Anim Iracl 24:467-476.
47. Schaafsma IA, van Imsl MG, Kooislra HS, VerkIei| C, Ieelers MI, oer I, Ri|nberk A, Iverls
MI 2002 Ixercise-induced hyerkaIemia in hyolhyroid dogs. Domesl Anim IndocrinoI
22:113-125.
GeneraI Inlroduclion
_________________________________________________________________________________


43
48. udsberg SC, Moore GI, KIaenbach K 1993 Thyroxine-resonsive uniIaleraI foreIimb
Iameness and generaIized neuromuscuIar disease in four hyolhyroid dogs. } Am Vel Med
Ass 202:1859-1860.
49. DeIauche A}, Cuddon IA, IodeII M, Devoe K, IoveI HC, SheIlon GD 1998 NemaIine Rods in
canine myoalhies: 4 case reorls and Iileralure reviev. } Vel Inlern Med 12:424-430.
50. DoIiger S, DeIverdier M, More }, Longearl L, Regnier A, MagnoI }I 1995 HislochemicaI sludy
of culaneous mucins in hyolhyroid dogs Vel IalhoI 32:628-634.
51. renl GA, Larsen IR, Davies TI 2008 Hyolhyroidism and lhyroidilis. In: WiIIiams Texlbook
of IndocrinoIogy. 11
lh
edilion. Kronenberg HM, MeImed S, IoIonsky KS, Larsen IR (Ids).
IhiIadeIhia, IA: W Saunders. : 377-409.
52. Ianciera DL 1994 An echocardiograhic and eIeclrocardiograhic sludy of cardiovascuIar
funclion in hyolhyroid dogs } Am Vel Med Assoc 205: 996-1000.
53. eaIe KM, Ioomberg MS, van GiIder } el aI. 1992 CorreIalion of racing and reroduclive
erformance in greyhounds vilh resonse lo lhyroid funclion lesling } Am Anim Hos Assoc
28:263-269.
54. }ohnson C, OIivier N, Nachreiner R, MuIIaney T 1999 Iffecl of
131
I-induced hyolhyroidism
on indices of reroduclive femaIe funclion in aduIl maIe dogs. } Vel Inl Med 13:104-110.
55. Indrieri R}, WhaIen LR, Cardinel GH, HoIIigay TA 1987 NeuromuscuIar abnormaIilies
associaled vilh hyolhyroidism and Iymhocylic lhyroidilis in lhree dogs. } Am Vel Med
Assoc 190:544-548.
56. }aggy A, OIiver }I, Ierguson DC, Mahaffey IA, GIaus T}r 1994 NeuroIogicaI manifeslalions
of hyolhyroidism: a relroseclive sludy of 29 dogs } Vel Inl Med 8: 328-336.
57. Higgins MA, RossmeisI }H }r, Ianciera DL 2006 Hyolhyroid-associaled cenlraI veslibuIar
disease in 10 dogs: 1999-2005. } Vel Inlern Med 20:1363-1369.
58. Ierguson DC 1994 Udale on lhe diagnosis of canine hyolhyroidism. Vel CIin Norlh Am
SmaII Anim Iracl 24:515-540.
59. Wiersinga WM 2000 NonlhyroidaI iIIness. In: Werner & Ingbar's The Thyoid. 8
lh
edilion.
raverman LI and Uliger RD (Ids.). IhiIadeIhia, IA: Liincoll WiIIiams & WiIkins.
:281-95.
60. eaIe KM 1990 Currenl Diagnoslic Techniques for evaIualing lhyroid funclion in lhe dog Vel
CIin Norlh Am SmaII Anim Iracl 20:1429-1439.
61. IIIiol DA, King LG, Zerbe CA 1995 Thyroid hormone concenlralions in crilicaIIy iII canine
inlensive care alienls. } Vel Imerg Cril Care 5:17-23.
62. Schoeman }I, Herrlage MI 2008 Serum lhyrolroin, lhyroxine and free lhyroxine
concenlralions as rediclors of morlaIily in crilicaIIy iII uies vilh arvovirus infeclion: a
modeI for human aedialric crilicaI iIIness` Microbes Infecl10:203-207.
63. Torres SMI, McKeever I}, }ohnslon SD 1991 Iffecl of oraI adminislralion of rednisoIone on
lhyroid funclion in dogs. Am } Vel Res 52:416-421.
64. Daminel S, Iaradis M, RefsaI KR, Irice C 1999 Shorl lerm infIuence of rednisoIon and
IhenobarbilaI on lhyroid funclion in eulhyroid dogs. Can Vel } 40:411-415.
Chaler 2
_________________________________________________________________________________

44

65. Daminel S, Ierguson DC 2003 InfIuence of drugs on lhyroid funclion in dogs. } Vel Inlern
Med 17:463-472.
66. Daminel S, CroubeIs S, Duchaleau L, Debunne A, van Geffen C, Hoybergs Y, van ree H, de
Rick A 2003 InfIuence of acelyIsaIicyIic acid and kelorofen on canine lhyroid funclion lesls.
Vel } 166:224-232.
67. HaII IA, CambeII KL, Chambers MD 1993 Iffecl of lrimelhorim/suIfamelhoxazoIe on
lhyroid funclion in dogs vilh yoderma, } Am Vel Med Assoc 202:1959-1962.
68. Ianciera DL 1990 Canine hyolhyroidism. Iarl II. Thyroid funclion. Tesls and lrealmenl.
Comend Conlin Iduc Iracl Vel 12:843-856.
69. NeIson RW, IhIe SL, IeIdman IC, olloms GD 1991 Serum free lhyroxine concenlralion in
heaIlhy dogs, dogs vilh hyolhyroidism, and eulhyroid dogs vilh concurrenl iIIness. } Am
Vel Med Assoc 198:401-1407.
70. Ielerson MI, MeIian C, NichoIs R 1997 Measuremenls of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroin concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Assoc. 211:1396-1402.
71. Torres SMI, Ieeney DA, Lekcharoensuk C, IIelcher TI, CIarkson CI, Nash NL, Hayden DW
2003 Comarison of coIIoid, lhyroid foIIicuIar eilheIium, and lhyroid hormone
concenlralions in heaIlhy and severeIy sick dogs. } Am Vel Med Assoc 222: 1079-1085.
72. Dixon RM, Mooney CT 1999 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40:72-78.
73. eIshav I, Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Amer Anim Hos Assoc 15:17-23.
74. Ri|nberk A 1996 IrolocoIs of funclion lesls. In: CIinicaI IndocrinoIogy of Dogs and Cals.
Ri|nberk (Id.). Dordrechl: KIuver Academic IubIishers. :205-212.
75. Sauve I, Iaradis M 2000 Use of recombinanl human lhyroid-slimuIaling hormone for
lhyrolrohin slimuIalion lesl in eulhyroid dogs. Can Vel } 41:215-219.
76. Roover De K, Duchaleau L, CarmichaeI N, van Geffen C, Daminel S 2006 Iffecl of slorage of
reconsliluled recombinanl human lhyroid-slimuIaling hormone (rhTSH) on lhyroid-
slimuIaling hormone (TSH) resonse lesling in eulhyroid dogs. } Vel Inl Med 20: 812-817.
77. orelli IS, Sieber-RucksluhI NS, WiIIi , Lulz H, Hofmann-Lehmann R, Reusch CI 2006
Comarison of lhe bioIogicaI aclivily of recombinanl human lhyroid-slimuIaling hormone
vilh bovine lhyroid-slimuIaling hormone and evaIualion of recombinanl human lhyroid-
slimuIaling hormone in heaIlhy dogs of differenl breeds. Am } Vel Res 67:1169-1172.
78. Daminel S, IifIe L, Iaradis M, Duchaleau L, and Moreau M 2007 Use of recombinanl human
lhyroid-slimuIaling hormone for lhyrolroin slimuIalion lesl in heaIlhy, hyolhyroid and
eulhyroid sick dogs. Can Vel } 48:1273-1279.
79. Nachreiner RI, Iorsberg M, }ohnson CA, RefsaI KR 1995 VaIidalion of an assay for canine
TSH (cTSH). ACVIM Abslracl 17. } Vel Inl Med 9:184.
80. WiIIiams DA, Scoll-Moncrieff }C, runer } 1995 Canine serum lhyroid-slimuIaling hormone
foIIoving induclion of hyolhyroidism. Abslracl 18. } Vel Inl Med 9:184.
GeneraI Inlroduclion
_________________________________________________________________________________


45
81. runer }M, Scoll-Moncrieff C, WiIIiams D 1996 DiurnaI fIuclualions of serum canine lhyroid
slimuIaling hormone (cTSH) in eulhyroid, hyolhyroid, and lhyroxine-suIemenled dogs.
ACVIM Abslracl 142. } Vel Inl Med 10:184.
82. Scoll-Moncrieff }C, NeIson RW, runer }M, WiIIiams DA 1996 Serum canine lhyrolroin
concenlralion (cTSH) in eulhyroid, hyolhyroid, and sick eulhyroid dogs. ACVIM Abslracl
151. } Vel Inl Med 10:186.
83. Ramsey IK, Ivans H, Herrlage MI 1997 Thyroid-slimuIaling hormone and lolaI lhyroxine
concenlralions in eulhyroid, sick eulhyroid and hyolhyroid dogs. } SmaII Anim Iracl.
38:540-545.
84. Scoll-Moncrieff }CR, NeIson RW 1998 Change in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolrohin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and dogs vilh concurrenl disease. } Am Vel Med Assoc 213:1435-
1438.
85. DanieI G, Shar DS, Nieckarz }A, Adams W 2002 Quanlilalive lhyroid scinligrahy as a
rediclor of serum lhyroxin concenlralion in normaI and hyerlhyroid cals. Vel RadioI
UIlrasound 43:374-382.
86. rmeI C, IoIIard RI, Kass IH, Samii VI, Davidson AI, RW NeIson 2005 UIlrasonograhic
evaIualion of lhe lhyroid gIand in heaIlhy, hyolhyroid, and eulhyroid GoIden Relrievers
vilh non lhyroidaI iIIness. } Vel Inlern Med 19:499-506.
87. Reese S, reyer U, Deeg C, Krafl W, Kaser 2005 Thyroid sonograhy as an effeclive looI lo
discriminale belveen eulhyroid sick and hyolhyroid dogs. } Vel Inlern Med 19:491-498.
88. Kemainen R}, Young DW, ehrend IN, CIark TI, SmiIey SD 1996 Auloanlibodies lo
lriiodolhyronine and lhyroxine in a goIden relriever } Am Anim Hos Assoc 32: 195-198.
89. Gunaralnam I 1986 The effecls of lhyroxine on hair grovlh in lhe dog } SmaII Anim Iracl.
27:17-29.
90. Mei| I 1997 TransshenoiaI hyohyseclomy for lrealmenl of iluilary-deendenl
hyeradrenocorlicism in dogs. Thesis Universileil Ulrechl.
91. Treier M and RosenfeId MG 1996 The hyolhaIamic-iluilary axis: co-deveIomenl of lvo
organs. Curr Oin CeII ioI 8:833-843.
92. Svanson LW 1987 The hyolhaIamus. In: Handbook of ChemicaI Neuroanalomy, Iarl 1.
|orkIun A, HokfeIl T and Svanson LW (Ids). Amslerdam: IIsevier IubIishing Comany.
:1-124.
93. HaImi NS, Ielerson MI, CoIurso G}, Liolla AS and Krieger DT 1981 Iiluilary inlermediale
Iobe in lhe dog: lvo ceII lyes and high bioaclive adrenocorlicolroin conlenl. Science 211:72-
74.
94. Kooislra HS, Greven SH, MoI }A, Ri|nberk A 1997 IuIsaliIe secrelion of o-meIanocyle-
slimuIaling hormone (o-MSH) by lhe ars inlermedia of lhe iluilary gIand and lhe
differenliaI effecls of dexamelhasone and haIoeridoI on lhe secrelion of o-MSH and
adrencorlicolrohic hormone in dogs. } Indocr 152:113-121.

Chaler 2
_________________________________________________________________________________

46

95. KeIIer-Wood MI and DaIIman MI 1984 Corlicosleroid inhibilion of ACTH secrelion.
Indocrine Rev 5:1-24.
96. WuIffraal NM, Drexhage HA, }eucken I, van der Gaag RD and Wiersinga WM 1987 Iffecls of
ACTH and ACTH fragmenls on DNA synlhesis in guinea-ig adrenaI segmenls kel in organ
cuIlure. } IndocrinoI 115:505-510.
97. AnlakIy T, Sasaki A, Liolla A, IaIkovils M and Krieger DT 1985 Induced exression of lhe
gIucocorlicoid recelor in lhe ral inlermediale iluilary Iobe. Science 229:277-279.
98. AnlakIy T, MerciIIe S and Cole }I 1987 Tisse-secific doaminergic reguIalion of lhe
gIucocorlicoid recelor in lhe ral iluilary. IndocrinoIogy 120:1558-1562.
99. IIolsky IM and VaIe W 1985 Iallerns of grovlh hormone-reIeasing faclor and somaloslalin
secrelion inlo lhe hyohysiaI-orlaI circuIalion of lhe ral. Science 230: 461-463.
100. Momany IA, ouvers CY, ReynoIds GA, Chang D, Hong A and NevIander K 1981 Design,
synlhesis and bioIogicaI aclivily of elides vhich reIease grovlh hormone, in vilro
IndocrinoIogy 108:31-39.
101. McKee KK, IaIyha OC, Ieighner SD, Hreniuk DL, Tan CI, IhiIis MS, Smilh RG, Van Der
IIoeg LH and Hovard AD 1997 MoIecuIar anaIysis of ral iluilary and hyolhaIamic grovlh
hormone secrelagogue recelors MoI IndocrinoI 11:415-423.
102. Ko|ima M., Hosoda H, Dale Y, Nakazalo M, Malsuo H and Kangava K 1999 GhreIin is a
grovlh hormone-reIeasing acyIaled elide from slomach. Nalure 402: 656-600.
103. halli SIM, Duchaleau L, Van Ham LML, De VIiegher SI, MoI }A, Ri|nberk A, Kooislra HS
2006 Iffecls of grovlh hormone secrelagogues on lhe reIease of adenohyohyseaI hormones
in young and oId heaIlhy dogs. Vel } 172:515-525.
104. MoI }A and Ri|nberk A 1997 Iiluilary funclion. In: CIinicaI iochemislry of Domeslic
AnimaIs. 5
lh
edilion. Kaneko }} (ed.). San Diego: Academic ress.: 517-551.
105. Iigenmann }I, Iigenmann RY, Ri|nberk A, Van Der Gaag I, Zaf } and Iroesch IR 1983
Irogeslerone-conlroIIed grovlh hormone overroduclion and naluraIIy occurring canine
diabeles and acromegaIy. Acla IndocrinoI 104:167-176.
106. SeIman I}, MoI }A, Rulleman GR, van Garderen I and Ri|nberk A 1994 Irogeslin-induced
grovlh hormone excess in lhe dog originales in lhe mammary gIand. IndocrinoIogy 134:287-
292.
107. Kooislra HS, den Herlog I, Okkens AC, MoI }A and Ri|nberk A 2000 IuIsaliIe secrelion
allern of grovlh hormone during lhe IuleaI hase and mid-anoeslrus in beagIe bilches. }
Rerod IerliIe 119:217-222.
108. Lee WM, Kooislra HS, MoI }A, DieIeman S}, Schaefers-Okkens AC 2006 Ovarieclomy during
lhe IuleaI hase infIuences secrelion of roIaclin, grovlh hormone, and insuIin-Iike grovlh
faclor-I in lhe bilch. TheriogenoIogy 66:484-490.
109. Ceda GI, Davis RG, RosenfeId RG and Hoffman AR 1987 The grovlh hormone (GH)-
reIeasing hormone (GHRH)-GH-somalomedin axis: evidence for raid inhibilion of GHRH-
eIiciled GH reIease by insuIin-Iike grovlh faclors I and II. IndocrinoIogy 120:1658-1662.
GeneraI Inlroduclion
_________________________________________________________________________________


47
110. IeIIegrini I, Iuel-Ia|ol MT, Mounier I, ennell I, Kordon C and IeIbaum } 1996 CenlraI
adminislralion of a grovlh hormone (GH) recelor mRNA anlisense increases GH uIsaliIily
and decreases hyolhaIamic somaloslalin exression in rals. } Neurosci 16:8140-8148.
111. Kooislra HS, Okkens AC, evers MM, Io-Sni|ders C, van Haaflen , DieIeman S},
Schoemaker } 1999 Concurrenl uIsaliIe secrelion of Iuleinizing hormone and foIIicIe-
slimuIaling hormone during differenl hases of lhe eslrous cycIe and aneslrus in beagIe
bilches. ioI Rerod 60:65-71.
112. De Gier }, Kooislra HS, D|a|adiningral-Laanen SC, DieIeman S}, Okkens AC 2006 DifferenliaI
reguIalion of lhe secrelion of Iuleinizing hormone and foIIicIe-slimuIaling hormone around
lhe lime of ovuIalion in lhe bilch. TheriogenoIogy 66: 1419-1422.
113. Shunik MA 1996 GonadaI Hormone feedback on iluilary gonadolroin genes. Trend
IndocrinoI Melab 7:272-276.
114. Vizcarra }A, Wellemann RI, raden TD, TurziIIo AM and Nell TM 1997 Iffecl of
gonadolroin-reIeasing hormone (GnRH) uIse frequency on serum and iluilary
concenlralions of Iuleinizing hormone and foIIicIe-slimuIalinhg hormone, GnRH recelors,
and messenger ribonucIeic acid for ganadolroin subunils in covs. IndocrinoIogy 138:594-
601.
115. Yu WH, Karanlh S, WaIczevska A, Sover SA and McCann SM 1997 A hyolhaIamic foIIicIe-
slimuIaling hormone-reIeasing decaelide in lhe ral. Iroc NalI Acd Sci USA 94:9499-9503.
116. en-}onalhan N 1985 Doamine: a roIaclin-inhibiling hormone. Indocr Rev 6:564-589.
117. Koch A, Hoen H-O, DieIeman S}, Kooislra HS, GunzeI-AeI A-R 2006 Iffecls of lhe
doamine agonisl cabergoIine on lhe uIsaliIe and TRH-induced secrelion of roIaclin, LH,
and lesloslerone in maIe beagIe dogs. TheriogenoIogy 65: 1666-1677.
118. Hinuma S, Habala Y, Iu|i R, Kavamala Y, Hosoya M, Iukusumi S, Kilada C, Masuo Y, Asano
T, Malsumolo H, Sekiguchi M, Kurokava T, Nishimura O, Onda H and Iu|ino M 1998 A
roIaclin-reIeasing elide in lhe brain. Nalure 393:272-276.
119. Sun , Iu|ivara K, Adachi S, Inoue K 2005 IhysioIogicaI roIes of roIaclin-reIeasing elide.
ReguI Iel 126:27-33.
120. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IM and
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114:275-282.
121. Kooislra HS, Okkens AC 2002 Secrelion of grovlh hormone and roIaclin during rogression
of lhe IuleaI hase in heaIlhy dogs: a reviev. MoI CeII IndocrinoI 197:167-172.
122. De Cosler R, eckers }I, eerens D and De Mey } 1983 A homoIogus radioimmunoassay for
canine roIaclin: Iasma IeveIs during lhe reroduclive cycIe. Acla IndocrinoI 103:473-478.
123. Overgaauv IA, Okkens AC, evers MM and Korlbeek LM 1998 Incidence of alenl Toxocara
infeclion in bilches during lhe oeslrous cycIe. Vel Quarl 20:104-107.
124. ehrends }, Irank K, Dogu I, rabanl G 1998 CenlraI nervous syslem conlroI of lhyrolroin
secrelion during sIee and vakefuIness. Horm Res 49:173-177.
Chaler 2
_________________________________________________________________________________

48

125. Negro-ViIar A, CuIIer MD, VaIenca MM, IIack T, Wisnievski G 1987 IuIsaliIe elide
secrelion: encoding of brain messages reguIaling endocrine and reroduclive funclions.
Inviron HeaIlh Iersecl 75:37-43.
126. rabanl G, Irank K and SchfI C 1992 IuIsaliIe allerns in hormone secrelion. Trends
IndocrinoI Melab 3: 183-190.
127. Kooislra HS 2000 AdenohyohyseaI funclion in heaIlhy dogs and in dogs vilh iluilary
disease. Thesis Universileil Ulrechl.
128. Merriam GR and Walcher KW 1982 AIgorilhms for lhe sludy of eisodic hormone secrelion
Am } IhysioI 243: I310-I318.
129. VeIdhuis }D, CarIson ML and }ohnson ML 1987 The iluilary gIand secreles in bursls:
araising lhe nalure of gIanduIar secrelory imuIses by simuIlaneous muIliIe-arameler
deconvoIulion of Iasma hormone concenlralions. Iroc NalI Acad Sci USA 84:7686-7690.


Chaptcr 3

Asscssmcnt nI Thyrnid Functinn in Dngs with
Lnw P!asma Thyrnxinc Cnnccntratinn









M.M. Diaz Espincira
1
, J.A. Mn!
1
, M.E. Pcctcrs
1
, Y.W.E.A. Pn!!ak
1
,
L. Ivcrscn
2
, J.E. van Di|k
3
, A. Ri|nbcrk
1
, and H.5. Knnistra
1


Jnurna! nI Vctcrinary Intcrna! Mcdicinc 2007, 21:25-32


1
Dearlmenl of CIinicaI Sciences of Comanion AnimaIs
IacuIly of Velerinary Medicine, Ulrechl Universily, Ulrechl, The NelherIands

2
CenlraI Laboralory of lhe Dearlmenl of CIinicaI Sludies, The RoyaI Velerinary
and AgricuIluraI Universily, Irederiksberg, Denmark

3
Dearlmenl of IalhobioIogy, IacuIly of Velerinary Medicine,
Ulrechl Universily, Ulrechl, The NelherIands.
Chapter 3
_____________________________________________________________________________
50

-

Differenlialion belveen hyolhyroidism and non-lhyroidaI iIIness in dogs
oses secific robIems because Iasma lolaI lhyroxine (TT4) concenlralions are
oflen Iov in non-lhyroidaI iIIness, and Iasma lhyroid slimuIaling hormone (TSH)
concenlralions are frequenlIy nol high in rimary hyolhyroidism. The serum
concenlralions of lhe common basaI biochemicaI variabIes (TT4, freeT4 |fT4j, and
TSH) overIa belveen dogs vilh hyolhyroidism and dogs vilh non-lhyroidaI
iIIness, bul, vilh slimuIalion lesls and quanlilalive measuremenl of lhyroidaI
99m
TcO4
-
ulake, differenlialion viII be ossibIe.
In 30 dogs vilh Iov Iasma TT4 concenlralion, lhe finaI diagnosis vas
based uon hisloalhoIogic examinalion of lhyroid lissue oblained by biosy.
Iourleen dogs had rimary hyolhyroidism, and 13 dogs had non-lhyroidaI
iIIness. Tvo dogs had secondary hyolhyroidism, and one dog had melaslalic
lhyroid cancer. The diagnoslic vaIue vas assessed of (1) Iasma concenlralions of
TT4, fT4, and TSH, (2) TSH-slimuIalion lesl, (3) Iasma TSH concenlralion afler
slimuIalion vilh TSH-reIeasing hormone (TRH), (4) occurrence of lhyrogIobuIin
anlibodies (TgAbs), and (5) lhyroidaI
99m
TcO4
-
ulake. IIasma concenlralions of
TT4, fT4, TSH, and lhe hormone airs TT4/TSH and fT4/TSH overIaed in lhe lvo
grous, vhereas, vilh TgAbs, lhere vas one faIse-negalive resuIl. ResuIls of lhe
TSH- and TRH-slimuIalion lesls did nol meel earIier eslabIished diagnoslic crileria,
overIaed, or bolh. Wilh quanlilalive measuremenl of lhyroidaI
99m
TcO4
-
ulake,
lhere vas no overIa belveen dogs vilh rimary hyolhyroidism and dogs vilh
non-lhyroidaI iIIness.
The resuIls of lhis sludy confirm earIier observalions lhal, in dogs,
accurale biochemicaI diagnosis of rimary hyolhyroidism oses secific
robIems. Irevious sludies, in vhich lhe TSH-slimuIalion lesl vas used as lhe
goId slandard for lhe diagnosis of hyolhyroidism may have suffered from
miscIassificalion. Quanlilalive measuremenl of lhyroidaI
99m
TcO4
-
ulake has lhe
highesl discriminalory over vilh regard lo lhe differenlialion belveen rimary
hyolhyroidism and non-lhyroidaI iIIness.




Assessmenl of lhyroid funclion
_____________________________________________________________________________


51

Intrnductinn

The basaI Iasma lolaI lhyroxine (TT4) concenlralion is beIov lhe reference
range in mosl dogs vilh rimary hyolhyroidism.
1-5
Hovever, lhe Iasma TT4
concenlralion of dogs vilh normaI lhyroid funclion can aIso be Iov due lo drugs
and iIIness. The lerms non-lhyroidaI iIIness and sick eulhyroid syndrome have
been inlroduced for lhis derangemenl of lhyroid hormone homeoslasis, as lhey
have in human medicine.
6,7
IIIness in lhis conlexl comrises virluaIIy aII non-
lhyroidaI diseases, surgicaI and non-surgicaI lrauma, and inadequale caIoric
inlake.
7
Drugs and iIIness have muIliIe effecls on lhyroid hormone secrelion,
lransfer, dislribulion, and melaboIism. These effecls incIude 1) decreased lhyroid
hormone binding lo Iasma carrier roleins, 2) increased melaboIic cIearance of
lhyroid hormones, 3) decreased T4 conversion lo lriiodolhyronine and increased T4
conversion lo melaboIicaIIy inaclive reverseT3, and 4) inhibilion of lhe secrelion of
TSH, lhyroid hormones, or bolh.
8
ConsequenlIy, lhe finding of a Iov basaI Iasma
TT4 concenlralion is of IillIe diagnoslic vaIue,
4,9
and, for lhis reason, slimuIalion
lesls by using eilher lhyroid slimuIaling hormone (TSH) or TSH-reIeasing
hormone (TRH) have been advocaled. Of lhese, lhe TRH-slimuIalion lesl vilh
measuremenl of lhe circuIaling TT4 concenlralion is nol abIe lo dislinguish
belveen dogs vilh hyolhyroidism and dogs vilh non-lhyroidaI iIIness.
5,10
Ior lhe
Iasl lvo decades, rimary hyolhyroidism in dogs has been diagnosed vilh a Iov
Iasma TT4 concenlralion lhal is nol resonsive lo slimuIalion vilh bovine TSH
(bTSH).
3-5,11,12

ecause lhis slimuIanl is no Ionger commerciaIIy avaiIabIe for diagnoslic
uroses in velerinary raclice, lhe diagnosis of rimary hyolhyroidism in dogs
is nov commonIy based on lhe combinalion of a Iov TT4 concenlralion and a high
TSH concenlralion.
2-4

Hovever, in as many as a lhird of dogs vilh rimary hyolhyroidism,
Iasma TSH concenlralions are vilhin lhe reference range.
2-4,13,14
Thus, in dogs, lhe
differenlialion belveen hyolhyroidism and non-lhyroidaI iIIness has become a
lhorny robIem, because of bolh lhe common occurrence of Iov Iasma TT4
concenlralions in non-lhyroidaI iIIness and lhe frequenl absence of a high Iasma
TSH concenlralion in rimary hyolhyroidism. Therefore, in dogs, hisloIogic
examinalion of lhyroid lissue oblained by biosy is regarded as lhe definilive and
mosl sensilive lesl lo idenlify lhyroid disease.
11,15
When lhe Iasma TT4
concenlralion is deressed in dogs vilh non-lhyroidaI iIIness, lhyroid hisloIogy is
normaI.
16
UnliI nov, lhere has been onIy one sludy of lhyrogIobuIin auloanlibodies
(TgAb) as diagnoslic variabIe in dogs in vhich lhe finaI diagnosis vas eslabIished
by lhyroid biosy.
17


Chapter 3
_____________________________________________________________________________
52

The urose of lhe resenl sludy vas lo assess lhe diagnoslic vaIue of (1)
Iasma concenlralions of TT4, free T4 (fT4), and TSH, (2) lhe TSH-slimuIalion lesl,
(3) changes in lhe Iasma TSH concenlralion afler slimuIalion vilh TRH, (4) lhe
occurrence of TgAb, and (5) radioaclive erlechnelale (
99m
TcO4
-
) ulake in lhe
lhyroid gIands in dogs resenled vilh symloms suggeslive of hyolhyroidism
and a Iov Iasma TT4 concenlralion. In lhese dogs, lhe finaI diagnosis of rimary
hyolhyroidism or non-lhyroidaI iIIness vas based uon hisloalhoIogic
examinalion of a lhyroid biosy.

Matcria!s and Mcthnds


The incIusion crileria for lhe dogs in lhis sludy vere (1) lhe resence of
signs comalibIe vilh hyolhyroidism, and (2) Iov Iasma TT4 concenlralion. Of
30 dogs vilh susicion of hyolhyroidism and a Iasma TT4 concenlralion (median
3 nmoI/I and range 2-18 nmoI/I, 0.23 and 0.16-1.4 g/dI) beIov lhe reference range
(19-46 nmoI/I, 1.5-3.6 g/dI), 14 had exlensive Ioss of lhyroid lissue on hisloIogicaI
examinalion of a lhyroid biosy. In 11 of lhese 14 dogs, lhere vas generaIized
infIammalory infiIlralion of Iymhocyles and occasionaIIy of Iasma ceIIs. In lhe
remaining 3, lhere vas comIele alrohy of lhe lhyroid lissue, vilhoul
infIammalory infiIlralion. The dogs' ages ranged from 2 lo 10 years (median 5).
Nine dogs vere maIe (3 caslraled) and 5 dogs vere femaIe (1 neulered). There vas
a vide variely of breeds. The median body veighl of lhe dogs vas 30 kg (range 5-
57 kg). IIeven dogs veighed more lhan 25 kg. Changes in coal and/or skin, such as
Ioss of coIor, lhinning of lhe hair coal, and aIoecia (n12), Ielhargy, and disinleresl
(n9) vere lhe mosl common resenling signs. In addilion, obesily or veighl gain
(n6), Iocomolion robIems (Iameness, sliff gail) (n3), and biIaleraI eriheraI
veslibuIar alaxia (n1) vere noliced. Al resenlalion, lhe lime from onsel of lhe
robIem(s) ranged from 5 monlhs lo 4 years. Afler comIelion of lhe sludies, lhe
dogs received (iniliaIIy) lvice daiIy 10 g lhyroxine er kg body veighl. Wilhin 14
days, aII dogs became more aIerl, aclive and resonsive, vilh lime lhere vas some
decrease in body veighl and comIele recovery of lhe hair coal.
In 13 dogs, hisloalhoIogic examinalion of lhe lhyroid biosy secimens
reveaIed normaI lhyroid gIand lissue, vilh lhe usuaI varialion in foIIicIe size. The
eilheIiaI ceIIs vere cubicaI, vilh some resorlion vacuoIes in lhe ad|acenl coIIoid.
Hence, lhe dogs vere cIassified as eulhyroid. Their ages ranged from 3 lo 11 years
(median, 6 years). Nine dogs vere maIe (3 caslraled) and 4 dogs vere femaIe (2
neulered). The median body veighl vas 39 kg (range, 7-56 kg). TveIve dogs
veighed more lhan 25 kg. Age and body veighl did nol differ significanlIy
belveen lhe dogs vilh rimary hyolhyroidism and lhose vilh non-lhyroidaI
iIIness. The variely of breeds vas aIso simiIar. Changes in coal and skin (n10),
Assessmenl of lhyroid funclion
_____________________________________________________________________________


53

Ielhargy (n8), and exercise inloIerance (n9) vere lhe mosl common signs. In
addilion, obesily or veighl gain (n5), increased aelile (n2), and oIyuria and
oIydisia (n1) vere menlioned al resenlalion. The lime from onsel aIso ranged
from 5 monlhs lo 4 years. IvenluaIIy, in lhese dogs lhe foIIoving diagnoses vere
made: obesily (n3), dermaloIogic condilions (n4, aloy, bacleriaI foIIicuIilis,
aIIergic dermalilis, ceIIuIilis), diIaled cardiomyoalhy (n1), chronic renaI faiIure
(n1), miId iluilary-deendenl hyeradrenocorlicism (n1), and no diagnosis
(n3).
In 2 dogs resenled vilh signs and symloms suggeslive of
hyolhyroidism and a Iov Iasma TT4 concenlralion, lhe hisloalhoIogicaI
diagnosis vas secondary hyolhyroidism. This vas based on lhe finding of fIal
foIIicuIar eilheIium in lhe absence of resorlion vacuoIes. In lhe remaining dog,
melaslalic lhyroid carcinoma vas found.

Tcsis
The adminislralion of medicalion vas sloed for a minimum of 6 veeks
before lhe lhyroid funclion sludies in 4 dogs. The eriod of lime eIased belveen
lhe firsl consuIlalion and lhe funclion lesls ranged from 2 lo 8 veeks. In each dog,
lhe rolocoI vas carried oul on lvo searale days, vilh al Ieasl 1 veek (range, 1-3
veeks) in belveen. On lhe firsl day, al 8:00 a.m. bIood samIes vere coIIecled by
cehaIic veniunclure for lhe delerminalion of lhe concenlralions of lolaI TT4,
TSH, fT4, and TgAb. ImmedialeIy lhereafler, quanlilalive ulake of
99m
TcO4
-
in lhe
neck area vas measured al 10, 20, 30, 45, 60, 90, and 120 minules afler in|eclion of
lhe radioharmaceulicaI. The TSH-slimuIalion lesl vas carried oul in lhe
aflernoon, and bIood samIes for Iasma TT4 measuremenls vere coIIecled in
hearin-coaled lubes immedialeIy before and 4 hours afler lhe inlravenous
adminislralion of 5 IU bTSH
a
(reference range for osl-TSH TT4, 39-80 nmoI/I |3.0-
6.2 g/dIj).
On lhe second day, a TRH-slimuIalion lesl vas erformed as arl of a
combined lesl of anlerior iluilary funclion.
18
Iood samIes vere coIIecled al 15,
0, 5, 10, 20, 30, and 45 minules afler inlravenous in|eclion of 10 g of TRH
b
er kg
body veighl and lransferred lo ice-chiIIed elhyIenediaminelelraacelic acid
(IDTA)-coaled lubes. IIasma vas cenlrifuged al 4C for 10 minules and slored al
-25C unliI assayed for TSH. Thereafler, a surgicaI lhyroid biosy for
hisloalhoIogic examinalion of lhyroid lissue vas laken.

Hcrncnc !cicrninaiicn
IIasma TT4 concenlralions vere measured vilh a homoIogous soIid-hase,
chemiIuminescenl enzyme immunoassay
c
according lo lhe inslruclions of lhe
manufaclurer and vaIidaled for dogs.
19
The inlra-assay coefficienls of varialion
Chapter 3
_____________________________________________________________________________

54
vere 14% and 8% al TT4 concenlralions of 8 and 25 nmoI/I (0.62 and 1.94 g/dI),
resecliveIy. The Iovesl deleclabIe amounl of TT4 vas 2 nmoI/I (0.16 g/dI). TT4
vaIues beIov 2 nmoI/I (0.16 g/dI) are exressed as 2 nmoI/I (0.16 g/dI).
Serum fT4 concenlralions vere measured vilh a commerciaI direcl diaIysis
kil
d
according lo lhe manufaclurer's inslruclions and as described reviousIy.
3,4

This direcl diaIysis melhod searales fT4 from bolh serum roleins and rolein-
bound T4 before measuring fT4 direclIy in lhe rolein-free diaIysale. The diaIysale
lhyroxine (fT4) is lhen measured vilh a sensilive, soIid-hase radioimmunoassay
(RIA) for T4. The inlra-assay coefficienls of varialion vere 11% and 12% al fT4
concenlralions of 19 moI/I (1.5 ng/dI) and 41 moI/I (3.2 ng/dI), resecliveIy. The
inler-assay coefficienls of varialion vere 15% al fT4 concenlralions of bolh 17
moI/I (1.3 ng/dI) and 35 moI/I (2.7 ng/dI). The Iovesl deleclabIe amounl of fT4
vas 1.9 moI/I (0.15 ng/dI). VaIues beIov lhis amounl are exressed as 1.9 moI/I
(0.15 ng/dI).
IIasma TSH concenlralions vere delermined vilh a homoIogous soIid-
hase, lvo-sile chemiIuminescenl enzyme immunomelric assay
e
according lo lhe
inslruclions of lhe manufaclurer, and as described reviousIy.
20
The inlra-assay
coefficienls of varialion vere 5%, 4%, and 4% al TSH concenlralions of 0.20, 0.50,
and 2.6 g/I, resecliveIy. The inlerassay coefficienls of varialion vere 6% and 8%
al TSH concenlralions of 0.16 and 2.8 g/I, resecliveIy. The Iovesl deleclabIe
amounl vas 0.03 g/I. The uer Iimil of lhe reference range for lhe Iasma TSH
concenlralion in eulhyroid dogs in our Iaboralory is 0.6 g/I. TSH vaIues beIov
0.03 g/I are exressed as 0.03 g/I.

Tnqrcg|c|u|in auicanii|c!ics
TgAbs vere delermined in serum samIes using an enzyme-Iinked
immunosorbenl assay.
17
The lesl resuIl for each samIe vas derived from olicaI
densily readings (OD) and exressed as a TgAb score (%) caIcuIaled from lhree in-
house caIibralors. The assay secificaIIy delecled TgAb, as |udged from lhe Iack of
resonse in lhe assay afler samIes had been incubaled vilh secific anligen. The
inlra- and inlerassay coefficienls of varialion ranged from 2.0 lo 4.9% and 4.6 lo
9.9%, resecliveIy. The deleclion Iimil vas an TgAb score of 5.6%. The cul-off
vaIue for a osilive resuIl vas 57.4%.
17
SamIes having an TgAb score higher lhan
lhe caIcuIaled cul-off vaIue vere considered osilive for TgAb, vhereas samIes
having an TgAb score equaI lo or Iover lhan lhe cul-off vaIue vere considered
negalive for TgAb.

99n
TcO4
-
upiakc
Scinligrahic imaging of lhe lhyroid gIands vas erformed vilh lhe
Inlegraled ORITIR Gamma Camera Syslem vilh Oen Icon Workslalion,
equied vilh a high resoIulion araIIeI-hoIe coIIimalor
f
. The dogs vere in|ecled
Assessmenl of lhyroid funclion
_____________________________________________________________________________


55

inlravenousIy vilh a dose of 111 lo 166 Mq radioaclive erlechnelale (
99m
TcO4
-
).
efore lhe inlravenous in|eclion, lhe radioaclivily in lhe fuII syringe vas measured
for 60 seconds in a osilion idenlicaI lo lhal of lhe dogs' lhyroids. The
adminislered
99m
TcO4
-
dose vas caIcuIaled by sublracling lhe radioaclivily in lhe
emly syringe.
The dogs vere manuaIIy reslrained in dorsaI recumbency. The fronl Iegs
vere slrelched caudaIIy aIong lhe lhorax, lhe head vas heId dovn on lhe bed, and
lhe coIIimalor vas osilioned above lhe venlraI surface of lhe neck. IIanar images
of lhe lhyroid gIands vere made and quanlilalive measuremenls of
99m
TcO4
-

ulake in lhe lhyroid gIands vere laken for 60 seconds (malrix, 256 x 256) al 5, 10,
20, 30, 45, 60, 90, and 120 minules afler lhe adminislralion of lhe
radioharmaceulicaI.
Ior lhe caIcuIalions lhe camera vas connecled lo a comuler vilh lhe
Siemens ICON comuler syslem soflvare.
f
In each image, lhree regions of inleresl
vere dravn, one for lhe background and one around each lhyroid gIand. The
resuIls are exressed as ercenl
99m
TcO4
-
ulake in bolh lhyroid gIands.

Tnqrci! |icpsq
Ior lhyroid biosy, lhe dogs vere remedicaled by inlravenous in|eclion
of medelomidine
g
, and aneslhesia vas induced vilh roofoI
h
. AII dogs vere
inlubaled, and aneslhesia vas mainlained vilh isofIurane
i
vaorized in a nilrous
oxide/oxygen mixlure in a circIe brealhing syslem. Thyroid lissues vere coIIecled
by surgicaI reseclion of a samIe secimen from lhe Iefl lhyroid Iobe, by using a
venlraI midIine aroach. These incisionaI biosies al lhe caudaI oIe comrised
aboul 25% of lhe lhyroid lissue (remnanls). Thyroid lissue secimens vere fixed by
immersion in neulraI-buffered 10% formaIin, embedded in araffin, cul al 5 m,
and slained vilh hemaloxyIin and eosin.

Ana|qsis cj !aia
Dala for lhe dogs vilh rimary hyolhyroidism (n14) and lhe dogs vho
are eulhyroid (n13) vere incIuded in lhe slalislicaI anaIysis. Dala for lhe
remaining 3 dogs (2 dogs vilh secondary hyolhyroidism and lhe dog vilh
lhyroid lumor) vere nol incIuded in lhe slalislicaI anaIysis.
The KoImogorov-Smirnov lesl vas used lo lesl lhe normaIily of lhe dala.
ody veighl, age, and TgAb scores vere comared vilh Sludenl's i-lesl.
Differences belveen lhe olher variabIes of dogs vilh hyolhyroidism and dogs
vilh non-lhyroidaI iIIness vere comared vilh Mann Whilney-U lesl. Differences
belveen Iasma TSH concenlralions before and afler TRH adminislralion vere
assessed vilh lhe Iriedman lesl. SubsequenlIy, muIliIe comarisons vere
erformed using lhe Sludenl-Nevman-KeuIs lesl. The incremenl in Iasma TSH
Chapter 3
_____________________________________________________________________________
56

concenlralion afler TRH adminislralion vas defined as lhe Iasma TSH
concenlralion al l30 minules minus lhe mean basaI Iasma TSH concenlralion
(mean of l -15 minules and l0 minules). VaIues are exressed as means
slandard devialion (SD) or as median and range. I<0.05 vas considered
significanl.

Ethics nI cxpcrimcntatinn

The exerimenls of lhis sludy vere aroved by lhe IlhicaI Commillee of
lhe IacuIly of Velerinary Medicine, Ulrechl Universily.

Rcsu!ts

In lhe 14 dogs vilh rimary hyolhyroidism, lhe Iasma TT4 concenlralion
ranged from 2 nmoI/I lo 4 nmoI/I (0.160.31 g/dI), and lhe median vaIue vas 2
nmoI/I (0.16 g/dI). In lhe 13 dogs vilh non-lhyroidaI iIIness, lhe Iasma TT4
concenlralion ranged from 2 nmoI/I lo 18 nmoI/I (0.161.40 g/dI), and lhe median
vaIue vas 8 nmoI/I (0.62 g/dI). The Iasma TT4 concenlralions of lhe dogs vilh
rimary hyolhyroidism vere significanlIy (I0.002) Iover lhan lhose of lhe dogs
vilh non-lhyroidaI iIIness (Iig 1).
In lhe hyolhyroid grou (n12), lhe serum fT4 concenlralion ranged from
1.9 mI/I lo 10.7 moI/I (0.150.83 ng/dI), and lhe median vaIue vas 1.9 moI/I (0.15
ng/dI) (Iig 1). In 11 of lhese 12 dogs, lhe serum fT4 concenlralion vas beIov lhe
deleclion Iimil of lhe assay, vhereas, lhe serum fT4 concenlralion in lhe remaining
dog vas 10.7 moI/I (0.83 ng/dI). The Ialler dog vas ovned by a velerinary
sludenl, vho al a very earIy slage of lhe disease vondered vhelher lhe Ielhargy in
her dog couId be caused by hyolhyroidism. In summary, lhe hisloalhoIogic
examinalion of lhe lhyroid lissue from lhe Ialler dog reveaIed no normaI foIIicIes,
many smaII foIIicIes vilh cubic lo cyIindricaI eilheIium, diffuse infiIlralion vilh
Iymhocyles and Iasma ceIIs, and moderale fibrosis.
In lhe dogs vilh non-lhyroidaI iIIness (n9), lhe serum fT4 concenlralion
ranged from 1.9 moI/I lo 13.1 moI/I (0.151.02 ng/dI), and lhe median vaIue vas
9.3 moI/I (0.72 ng/dI). One of lhese 9 dogs had a serum fT4 concenlralion beIov
lhe deleclion Iimil of lhe assay. The serum fT4 concenlralions in lhe dogs vilh
rimary hyolhyroidism vere significanlIy (I0.001) Iover lhan lhose in lhe dogs
vilh lhe non-lhyroidaI iIIness syndrome (Iig 1).
The Iasma TSH concenlralion in lhe 14 dogs vilh hyolhyroidism ranged
from 0.03 lo 5 g/I, and lhe median vaIue vas 1.3 g/I. Iighl dogs had a Iasma
TSH concenlralion above lhe uer Iimil of lhe reference range (> 0.6 g/I), and 6
dogs had a Iasma TSH concenlralion vilhin lhe reference range (0.0-0.6 g/I).

Assessmenl of lhyroid funclion
_____________________________________________________________________________


57
















ox Iols of basaI circuIaling concenlralions of lolaI T4, free T4, and lhyroid slimuIaling
hormone in dogs vilh rimary hyolhyroidism and dogs vilh non-lhyroidaI iIIness.
OulIying dala oinls are reresenled by oen circIes.



The Iasma TSH concenlralions in 12 of lhe 13 dogs vilh non-lhyroidaI iIIness
vere vilhin lhe reference range, and ranged from 0.03 lo 0.98 g/I (median, 0.2
g/I). The Iasma TSH concenlralions in lhe dogs vilh rimary hyolhyroidism
vere significanlIy (I0.01) higher lhan lhose in lhe dogs vilh lhe non-lhyroidaI
iIIness (Iig 1).
No adverse reaclions vere noliced in any of lhe dogs afler inlravenous
adminislralion of bTSH. The basaI Iasma TT4 concenlralion vas s 5 nmoI/I (0.4
g/dI) in aII 14 dogs vho vere hyolhyroid. Iour hours afler adminislralion of
bTSH, lhe Iasma TT4 concenlralion had nol increased in 7 of lhese dogs, and onIy
sIighlIy in lhe remaining 7 dogs vho vere hyolhyroid (lo s8 nmoI/I |0.62 g/dIj).
The incremenl of lhe Iasma TT4 concenlralion in lhe dogs vho vere hyolhyroid
ranged from 0 lo 5 nmoI/I (00.4 g/dI), median, 0.5 nmoI/I (0.04 g/dI). The
incremenl of lhe Iasma TT4 concenlralion in lhe dogs vilh non-lhyroidaI iIIness
ranged from 0 lo 29 nmoI/I (02.3 g/dI), vilh a median vaIue of 16 nmoI/I (1.2
g/dI). In 1 dog, lhe Iasma TT4 concenlralion did nol increase afler adminislralion
of bTSH. The TT4 incremenl afler bTSH slimuIalion vas significanlIy higher
(I<0.001) in lhe dogs vilh non-lhyroidaI iIIness lhan in lhe dogs vilh rimary
hyolhyroidism.
Chapter 3
_____________________________________________________________________________

58
TRH adminislralion did nol cause adverse reaclions in any of lhe dogs. In
bolh lhe dogs vilh hyolhyroidism and lhe dogs vilh non-lhyroidaI iIIness, lhe
Iasma TSH concenlralion increased significanlIy (I<0.0001) afler TRH
adminislralion (Iig 2). In lhe dogs vilh non-lhyroidaI iIIness, lhe TSH curve
eaked al 30 minules (Iig 2). In 6 dogs vilh hyolhyroidism, lhe maximaI
incremenl in Iasma TSH concenlralion afler TRH adminislralion vas Iess lhan 0.1
g/I, vhereas lhis occurred in onIy 1 dog vilh non-lhyroidaI iIIness. Al 30 minules
afler TRH adminislralion, Iasma TSH concenlralions did nol differ significanlIy
belveen grous. Desile lhis overIa, in lhe dogs vilh hyolhyroidism, lhe
incremenl of Iasma TSH concenlralion al 30 min afler TRH adminislralion
(median 0.04 g/I, range, 0 lo 1.15 g/I) vas significanlIy (I0.001) Iover lhan lhal
in lhe dogs vilh non-lhyroidaI iIIness (median, 0.36 g/I, range 0.05 lo 1.37 g/I.
The TgAb score (mean SD, 581% 609%) vas delermined in 5 dogs vilh
hyolhyroidism. One dog vilh hyolhyroidism vas negalive for TgAb.
HisloalhoIogic examinalion of lhe lhyroid biosy of lhis dog reveaIed end-slage
Iymhocylic lhyroidilis. HisloalhoIogic examinalion of lhe lhyroid biosy
secimens from lhe olher 4 dogs reveaIed Iymhocylic lhyroidilis. The TgAb score
(19 4%) vas delermined in 6 dogs vilh non-lhyroidaI iIIness, and vas negalive in
aII of lhem. The mean TgAb score in lhe dogs vilh rimary hyolhyroidism vas
significanlIy (I<0.001) higher lhan lhal in lhe dogs vilh non-lhyroidaI iIIness.
In lhe erlechnelale ulake sludies, lhere vas no overIa in lhyroid
99m
TcO4
-
ulake 45-120 minules afler radioharmaceulicaI in|eclion belveen lhe
dogs vilh rimary hyolhyroidism and lhe dogs vilh non-lhyroidaI iIIness (Iig 3).
In lhe dogs vilh hyolhyroidism, ulake al 60 minules ranged from 0.03 lo 0.26%
of lhe in|ecled dose (median, 0.16%) and, in lhe dogs vilh non-lhyroidaI iIIness,
from 0.39 lo 1.86% of lhe in|ecled dose (median, 0.62%).
LaslIy ve caIcuIaled ralios of lhe hormone airs TT4/TSH and fT4/TSH. In
lhe dogs vilh hyolhyroidism and lhe dogs vilh non-lhyroidaI iIIness lhe ralios
ranged from 0.6 lo 66.6 (median, 2.5) and from 3.8 lo 433 (median, 40.7),
resecliveIy. Ior lhe fT4/TSH ralio lhese figures vere 0.763.3 (median, 2.9) and 10
410 (median, 34.4). In lhe lvo dogs vilh secondary hyolhyroidism lhe TT4/TSH
ralios (66.6 and 42.8) and lhe fT4/TSH ralios (63.3 and 27.1) vere in lhe range of
lhose of bolh lhe dogs vilh hyolhyroidism and lhe dogs vilh non-lhyroidaI
disease.




Assessmenl of lhyroid funclion
_____________________________________________________________________________


59





































Mean (and SD) Iasma TSH concenlralion in 14 dogs vilh rimary hyolhyroidism (oen
circIes) and 13 dogs vilh non-lhyroidaI iIIness (cIosed circIes) as infIuenced by inlravenous
adminislralion of TRH (al lime zero).

Chapter 3
_____________________________________________________________________________

60



































Median vaIues and ranges for lhyroidaI ulake of
99m
TcO4
-
, measured as ercenl ulake of
in|ecled dose, in 14 dogs vilh rimary hyolhyroidism and 13 dogs vilh non-lhyroidaI
iIIness.
Assessmenl of lhyroid funclion
_____________________________________________________________________________


61



In lhis sludy, differenlialion belveen hyolhyroidism and non-lhyroidaI
iIIness vas based uon hisloalhoIogic examinalion of lhyroid lissue. AIlhough
lhis is regarded as lhe definilive and mosl sensilive lesl lo idenlify lhyroid
Iesions,
11,15
lhe resence of a Iesion does nol necessariIy mean lhe resence of a
funclionaI abnormaIily.
21
Therefore, in lhis sludy, lhe funclionaI slalus of lhe
lhyroid gIand vas examined by ulake of
99m
TcO4
-
. The ulake of lracer vas
very Iov in lhe dogs vilh lhyroid Iesions comalibIe vilh rimary
hyolhyroidism and cIearIy differenl from lhe ulakes seen in lhe dogs vilh non-
lhyroidaI disease. Thus, lhe diagnosis of hyolhyroidism and non-lhyroidaI
disease vas eslabIished morhoIogicaIIy and funclionaIIy.
The hisloIogicaIIy characlerized Iesions in lhe dogs vilh hyolhyroidism
comrised end-slage Iymhocylic lhyroidilis and comIele alrohy. In lhis
immune-medialed lhyroidilis, lhe deslruclion of lhe lhyroid gIand occurs in
associalion vilh infIammalory ceII infiIlrales consisling of Iymhocyles and
Iasma ceIIs vilh occasionaI macrohages.
11,12
The rocess rogresses lo an end-
slage form, characlerized by reIacemenl of lhyroid arenchyma by roIiferaling
fibrous conneclive lissue, some degeneraling foIIicuIar ceIIs, and scallered
infIammalory ceIIs,

as found in some of lhe dogs vilh hyolhyroidism of lhe
resenl sludy. In comIele alrohy, aIso caIIed idioalhic lhyroid alrohy, lhere
are no ceIIuIar infiIlrales. The alhogenesis has been suggesled lo be differenl, bul,
in humans, lhis form is nov being considered lhe end slage of auloimmune
lhyroid disease.
23

Thyroxine is lhe main secrelory roducl of lhe lhyroid gIand, and,
lherefore, lhe Iasma TT4 concenlralion is lhe mosl aroriale variabIe lo
measure in dogs susecled of hyolhyroidism. In lhe resenl sludy, aII dogs vilh
(hisloIogicaIIy roven) hyolhyroidism had very Iov TT4 concenlralions,
consislenl vilh revious sludies reorling lhal TT4 measuremenl is a sensilive lesl
for diagnosing hyolhyroidism.
1-4, 24
Il has been reorled, hovever, lhal in some
dogs vilh rimary hyolhyroidism, lhe Iasma TT4 concenlralion may exceed lhe
Iover Iimil of lhe reference range,
3,5
aIlhough, in lhese cases, lhe diagnosis vas nol
eslabIished by hisloalhoIogic examinalion. CircuIaling anlibodies direcled againsl
T4 may resuIl in a faIseIy eIevaled Iasma TT4 concenlralion in hyolhyroid dogs.
25

In lhese dogs vilh non-lhyroidaI iIIness, lhe Iasma TT4 concenlralions
vere significanlIy higher lhan lhose in lhe dogs vilh rimary hyolhyroidism.
Hovever, some of lhe dogs vilh non-lhyroidaI iIIness iniliaIIy had very Iov
Iasma TT4 concenlralions. This confirms lhal, vhiIe lhe sensilivily of TT4
measuremenls in diagnosing hyolhyroidism is high, lhe secificily is Iov.
4,9
The
overIa in lhe Iasma TT4 concenlralion in lhe lvo condilions may easiIy resuIl in
Chapter 3
_____________________________________________________________________________
62

an erroneous diagnosis of hyolhyroidism and consequenlIy may Iead lo
inaroriale (IifeIong) lrealmenl. Il vas inleresling lo see lhal, in lhe dogs vilh
non-lhyroidaI iIIness, lhe basaI TT4 concenlralions in lhe TSH-slimuIalion lesl vere
higher lhan lhe iniliaI concenlralions al firsl resenlalion. One mighl secuIale lhal
some of lhe dogs al firsl examinalion vere sliII under lhe infIuence of some
medicalion lhal couId Iover TT4. In lhe eriod eIased belveen lhe firsl
examinalion and lhe aoinlmenl for furlher sludies, lhis effecl mighl have
diminished.
The fT4 concenlralions lended lo be more secific lhan lhe Iasma TT4
concenlralion. There vas IillIe overIa of fT4 vaIues belveen non-lhyroidaI iIIness
and hyolhyroidism. Hovever, lhe sensilivily of fT4 measuremenls seemed Iover
lhan lhal of TT4 measuremenls. In agreemenl vilh an earIier reorl,
4
ve found one
dog vilh incomIele deslruclion of lhyroid lissue in vhich lhe TT4 concenlralion
vas abnormaIIy Iov, vhereas lhe fT4 concenlralion vas nol. The sIighlIy beller
secificily of fT4 comared vilh TT4 couId be considered a reason lo incIude lhis
lesl in rouline diagnoslics for hyolhyroidism,
3,5
afler eslabIishmenl of a reference
range in a Iarge and aroriale number of dogs.
Ior lhe Iasl 20 years, lhe diagnosis of canine rimary hyolhyroidism has
been based on a Iov TT4 concenlralion lhal does nol resond afler 4 or 6 hours
afler TSH slimuIalion.
3,4,11
This lesl has aIso been used as lhe goId slandard for
dislinguishing dogs vilh hyolhyroidism from dogs vilh non-lhyroidaI iIIness,
aIlhough il is recognized lhal, in dogs vilh non-lhyroidaI iIIness, lhe resuIls of TSH
slimuIalion lesl may mimic lhose of dogs vilh hyolhyroidism.
15
Indeed, in some
of lhese dogs vilh non-lhyroidaI iIIness, ve found onIy a minor increase in lhe
Iasma TT4 concenlralion. If ve used earIier cul-off vaIues for hyolhyroidism
(osl-slimuIalion T4 > 25 nmoI/I and al Ieasl 1.5 limes lhe baseIine concenlralion),
3

some dogs vilh non-lhyroidaI iIIness vouId have been miscIassified as dogs vilh
rimary hyolhyroidism. Il is lhus IikeIy lhal lhis miscIassificalion has aIso
occurred in revious sludies assessing olher lesls, and lhis shouId be borne in
mind vhen evaIualing lhe resuIls of lhese sludies. Desile lhis, lhe TSH-
slimuIalion lesl may sliII be regarded as lhe Ieasl inconcIusive biochemicaI lhyroid
funclion lesl for differenlialing belveen rimary hyolhyroidism and lhe non-
lhyroidaI iIIness syndrome in dogs. Ior some lime, lhe unavaiIabiIily of bTSH has
Iimiled lhe use of lhe TSH-slimuIalion lesl, bul, recenlIy, il has become cIear lhal
recombinanl human lhyroid-slimuIaling hormone (rhTSH) can be used as an
aIlernalive, vilhoul adverse reaclions.
26
AIso in lhis aroach, reduced slimuIalion
can be found in dogs vilh concurrenl disease and medicalion.
|

The diagnosis of rimary hyolhyroidism in dogs is currenlIy oflen based
on lhe combinalion of a Iov TT4 concenlralion and a high TSH concenlralion in a
singIe Iasma samIe. In Iine vilh revious reorls,
2-4,13,14
ve found a reIaliveIy
high number of dogs vilh rimary hyolhyroidism lo have Iasma TSH
Assessmenl of lhyroid funclion
_____________________________________________________________________________


63

concenlralion vilhin lhe reference range for eulhyroid dogs. SeveraI exIanalions
for lhis henomenon have been suggesled, such as concurrenl secondary
hyolhyroidism,
2,13
faiIure of lhe TSH assay lo delecl aII isoforms of circuIaling
TSH,
3
decrease in reviousIy high TSH vaIues in dogs vilh hyolhyroidism vilh
lime,
27
and uIlradian fIuclualions in lhe circuIaling concenlralion of TSH.
20

The basaI Iasma TSH concenlralion in lhe dogs vilh non-lhyroidaI iIIness
vas vilhin lhe reference range, vilh lhe excelion of one dog vhich had a high
Iasma TSH concenlralion. The occurrence of Iasma TSH concenlralions
exceeding lhe reference range has been reorled before in dogs vho are
eulhyroid.
2-4,13
Il has aIso been reorled lhal lhe circuIaling TSH concenlralion in
dogs vilh non-lhyroidaI iIIness is significanlIy higher lhan lhal in heaIlhy
conlroIs.
9
These high concenlralions of circuIaling TSH in lhe non-lhyroidaI iIIness
syndrome may be ascribed lo changes in lhe uIsaliIe nalure of TSH secrelion or lo
recovery from a lransienl hyolhyroid slale, or bolh, bul lhe generaI concIusion
seems lo be lhal, in bolh humans and dogs vilh non-lhyroidaI iIIness, TSH
concenlralions remain vilhin lhe reference range in mosl cases.
9,28

Measuremenl of lhe TSH resonse afler TRH adminislralion offers a raid,
safe, and effeclive melhod of invesligaling lhe reserve caacily of iluilary
lhyrolrohic ceII funclion.
18,29
TheorelicaIIy, lhe TRH-slimuIalion lesl may aIso be
used in lhe diagnosis of rimary hyolhyroidism, because exogenous TRH may acl
as an amIifier lo exaggerale an abnormaIily in TSH secrelion.
30
Indeed,
exerimenlaIIy induced smaII decreases in Iasma T4 concenlralions cause
increased basaI TSH concenlralions, as veII as an increased resonse lo TRH.
31

Hovever, in dogs vilh rimary hyolhyroidism, lhere is no exaggeraled TSH
resonse lo slimuIalion vilh TRH. On lhe conlrary, in Iine vilh revious
sludies,
2,32
ve found lhe increase lo be significanlIy smaIIer lhan lhal in lhe dogs
vilh non-lhyroidaI iIIness. Moreover, in aImosl haIf of lhe dogs vilh
hyolhyroidism, lhe increase vas Iess lhan 0.1 g/I. This indicales lhal iluilary
resonsiveness lo TRH is considerabIy decreased in dogs vilh rimary
hyolhyroidism. Concurrenl secondary hyolhyroidism
2,13
and reduced TSH
gIycosyIalion resuIling in faiIure of lhe TSH assay lo delecl aII isoforms of
circuIaling TSH
3
have been suggesled as exIanalions for lhe reIaliveIy Iov Iasma
TSH concenlralions in dogs vilh hyolhyroidism and may conlribule lo lhe
diminished TSH resonse uon TRH slimuIalion.
The absoIule increase in Iasma TSH concenlralion afler TRH
adminislralion in lhe dogs vilh non-lhyroidaI iIIness vas Iess lhan lhal reorled in
heaIlhy beagIe dogs.
18
AIso, humans vilh non-lhyroidaI iIIness oflen have a Iess-
lhan-normaI TSH resonse lo TRH slimuIalion,
33
vhich shouId be seen as a
consequence of lhe disease in a alienl vho is sliII eulhyroid. We did nol find lhe
Iasma TSH concenlralions al 30 minules afler TRH adminislralion lo differ
Chapter 3
_____________________________________________________________________________
64

significanlIy belveen lhe lvo grous of dogs. This recIudes lhe use of lhis lesl lo
dislinguish belveen rimary hyolhyroidism and lhe non-lhyroidaI iIIness
syndrome, as found earIier.
30,32

The TgAb score vas osilive in 4 of 5 dogs vilh rimary hyolhyroidism
because of Iymhocylic lhyroidilis. In lhe dog vilh rimary hyolhyroidism vho
had a negalive TgAb score, hisloalhoIogic examinalion of lhe lhyroid biosy
reveaIed end-slage Iymhocylic lhyroidilis. In lhe absence of lhyroid anligen,
lhere vas robabIy no slimuIus for lhe formalion of anlibodies againsl
lhyrogIobuIin.
5,17,34
CircuIaling TgAbs may be absenl in humans vilh Iong-slanding
rimary hyolhyroidism.
35

The TgAb score vas negalive in aII lhe lesled dogs vilh non-lhyroidaI
iIIness. The absence of faIse-osilive resuIls may have been because of roer
cIassificalion, aIlhough il is veII knovn lhal circuIaling TgAbs may occur in dogs
vho are eulhyroid. There is generaI agreemenl lhal lhe resence of circuIaling
TgAbs indicales lhe resence of lhyroidilis, bul lhal il is nol roof lhal
hyolhyroidism is resenl or viII deveIo.
17,36,37
AIlhough lhe occurrence of TgAbs
in heaIlhy individuaIs Iimils lhe diagnoslic vaIue of lhis lesl, a osilive TgAb score
in a dog vilh cIinicaI signs consislenl vilh hyolhyroidism and a Iov Iasma TT4
concenlralion increases lhe IikeIihood lhal lhis dog is suffering from
hyolhyroidism from (earIy slages) of Iymhocylic lhyroidilis.
In endocrine disease, lhe eriheraI hormone and lhe reguIaling
(iluilary) hormone oflen change in oosile direclions, as a resuIl of lhe feedback
syslem. Ior lhis reason, caIcuIalion of lhe ralio of such hormone airs may
imrove lhe discriminalion belveen diseased and non-diseased animaIs, as has
been demonslraled in lhe diagnosis of cals vilh rimary hyeraIdosleronism and
in dogs vilh rimary hyoadrenocorlicism.
38,39
Hovever, ve found lhal
caIcuIalion of lhe TT4/TSH and lhe fT4/TSH ralios did nol imrove diagnoslic
accuracy. The overIa belveen lhe dogs vilh hyolhyroidism and lhe dogs vilh
non-lhyroidaI disease ersisled, IargeIy as a resuIl of lhe absenl or onIy minor rise
in lhe Iasma TSH concenlralion in lhe dogs vilh hyolhyroidism in resonse lo
lhe Iovering of lhe Iasma lhyroxine concenlralion.
CaIcuIalion of lhe ercenl ulake of
99m
TcO4
-
has roven lo be very usefuI
in lhe assessmenl of lhyroid funclion in humans and cals.
40,41
We found lhis aIso lo
be lhe case for dogs vilh hyolhyroidism.
99m
TcO4
-
ulake in lhe dogs vilh
rimary hyolhyroidism and lhe dogs vilh non-lhyroidaI iIIness vas differenl
belveen 45 and 120 minules afler lracer in|eclion. In lhe dogs vilh non-lhyroidaI
iIIness, lhe
99m
TcO4
-
ulake vas comarabIe lo lhal reorled for heaIlhy beagIe
dogs.
42
Il vas inleresling lo see lhal lhe dog in an earIy slage of hyolhyroidism
had
99m
TcO4
-
ulake in lhe uer arl of lhe range for lhe dogs vilh
hyolhyroidism, bul did nol have lhe highesl ulake vaIue.

Assessmenl of lhyroid funclion
_____________________________________________________________________________


65

In concIusion, Iasma concenlralions of commonIy measured hormones
(TT4, fT4, TSH) overIaed in lhe dogs vilh rimary hyolhyroidism and lhe dogs
vilh non-lhyroidaI disease, vhereas TgAb scores gave one faIse-negalive resuIl. In
lhe lvo grous of dogs, lhe resuIls of bolh lhe TSH- and TRH-slimuIalion lesls did
nol meel earIier eslabIished diagnoslic crileria, overIaed or bolh, indicaling lhal
assessmenl sludies by using lhe TSH-slimuIalion lesl as lhe goId slandard for
lhe diagnosis of hyolhyroidism may have suffered from miscIassificalion.
Quanlilalive measuremenl of
99m
TcO4
-
had lhe highesl discriminalory over, lhere
being no overIa belveen dogs vilh rimary hyolhyroidism and dogs vilh non-
lhyroidaI iIIness.

Fnntnntcs

a
bTSH, Ierring V, Hoofddor, The NelherIands
b
TRH, Hoffmann-La Roche, aseI, SvilzerIand
c
ImmuIile canine TolaI T4, DIC, Los AngeIes, CA
d
Iree T4 by equiIibrium diaIysis, NichoIs Inslilule Diagnoslics, San }uan
Caislrano, CA
e
ImmuIile canine TSH, DIC, Los AngeIes, CA
f
Siemens MedicaI Syslems, The NelherIands
g
Domilor, SmilhKIine eecham AnimaI HeaIlh V, Zoelermeer, The NelherIands
h
Dirivan, Zeneca V, Ridderkerk, The NelherIands
i
Iorene, Abboll Laboralories V, Maarsen, The NelherIands
|
orelli IS, Sieber-RucksluhI NS, Hofmann-Lehmann R el aI. Use of recombinanl
human TSH in TSH-slimuIalion lesls in susecled hyolhyroid dogs. ACVIM
abslracl 91. } Vel Inlern Med 2005, 19: 424 (abslracl)


Acknnw!cdgmcnts

The aulhors lhank Mrs. M.I. van WoIferen, and Mr. H.G.H. van IngeIen,
for lheir skiIIed and enlhusiasl lechnicaI assislance.
Chapter 3
_____________________________________________________________________________

66



1. eIshav I, Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Am Anim Hos Assoc 15:17-23.
2. Ramsey IK, Ivans H, Herrlage MI 1997 Thyroid-slimuIaling hormone and lolaI lhyroxine
concenlralions in eulhyroid, sick eulhyroid and hyolhyroid dogs. } SmaII Anim Iracl 38:540-
545.
3. Ielerson MI, MeIian C, NichoIs R 1997 Measuremenls of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroin concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Assoc 211:1396-1402.
4. Dixon RM, Mooney CT 1999 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40:72-78.
5. Kemainen R}, ehrend IN 2001 Diagnosis of canine hyolhyroidism. Ierseclives from a
lesling Iaboralory. Vel CIin Norlh Am SmaII Anim Iracl 31:951-62.
6. Daminel S, Ierguson DC 2003 InfIuence of drugs on lhyroid funclion in dogs. } Vel Inlern
Med 17:463-472.
7. Wiersinga WM 2000 NonlhyroidaI iIIness. In: Werner and Ingbar's The Thyoid. 8
lh
edilion.
IhiIadeIhia, IA: Liincoll WiIIiams & WiIkins.:281-295.
8. Kalein IM, Hays MT, Ierguson DC 1994 Thyroid hormone melaboIism. A comaralive
evaIualion. Vel CIin Norlh Am SmaII Anim Iracl 24:431-461.
9. Kanlrovilz L, Ielerson MI, MeIian C, NichoIs R 2001 Serum lolaI lhyroxine, lolaI
lriiodolhyronine, free lhyroxine, and lhyrolrohin concenlralions in dogs vilh nonlhyroidaI
disease. } Am Vel Med Assoc 219:765-769.
10. Irank LA 1996 Comarison of lhyrolroin-reIeasing hormone (TRH) lo lhyrolroin (TSH)
slimuIalion for evaIualing lhyroid funclion in dogs. } Am Anim Hos Assoc 32:481-487.
11. eIshav I 1983 Thyroid disease. In: Texlbook of Velerinary InlernaI Medicine, Diseases of
lhe Dog and lhe Cal. 2
nd
edilion. Illinger S} (Id). IhiIadeIhia, IA: W Saunders. :1592-
1614.
12. Scoll-Moncrieff }CR and GuliII-Yoran L 2000 Hyolhyroidism. In: Texlbook of Velerinary
InlernaI Medicine, Diseases of lhe Dog and lhe Cal. 5
lh
edilion. Illinger S} and IeIdman IC
(Ids). IhiIadeIhia, IA: W Saunders. :1419-1429.
13. Scoll-Moncrieff }CR, NeIson RW, runer }M, WiIIiams DA 1998 Comarison of serum
concenlralions of lhyroid-slimuIaling hormone in heaIlhy dogs, hyolhyroid dogs, and
eulhyroid dogs vilh concurrenl disease. } Am Vel Med Assoc 212:387-391.
14. }ensen AL, Iversen L, Hoier R, el aI. 1996 IvaIualion of an immunoradiomelric assay for
lhyrolroin in serum and Iasma samIes of dogs vilh rimary hyolhyroidism. } Com
Ialh 114:339-346.
15. IeIdman IC and NeIson RW 2004 Hyolhyroidism. In: Canine and IeIine IndocrinoIogy and
Reroduclion. 3
rd
edilion. IhiIadeIhia, IA: W Saunders. :85-151.

Assessmenl of lhyroid funclion
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67

16. Torres SMI, Ieeney DA, Lekcharoensuk C, el aI. 2003 Comarison of coIIoid, lhyroid
foIIicuIar eilheIium, and lhyroid hormone concenlralions in heaIlhy and severeIy sick dogs. }
Am Vel Med Assoc 222:1079-1085.
17. Iversen L, }ensen AL, Hoier R, el aI. 1998 DeveIomenl and vaIidalion of an imroved
enzyme-Iinked immunosorbenl assay for lhe deleclion of lhyrogIobuIin auloanlibodies in
canine serum samIes. Domesl Anim IndocrinoI 15:525-536.
18. Mei| I, MoI }A, Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13:465-468.
19. runer }M, Scoll-Moncrieff CR, WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am
Vel Med Assoc 212:1572-1575.
20. Kooislra HS, Diaz Isineira MM, MoI }A, el aI. 2000 Secrelion allern of lhyroid-slimuIaling
hormone in dogs during eulhyroidism and hyolhyroidism. Dom Anim IndocrinoI 18:19-29.
21. Graham IA, Nachreiner RI, RefsaI KR, Irovencher-oIIiger AL 2001 Lymhocylic lhyroidilis.
Vel CIin Norlh Am SmaII Anim Iracl 31:915-933.
22. GosseIin S}, Caen CC, Marlin SL 1981 HisloIogic and uIlraslrucluraI evaIualion of lhyroid
Iesions associaled vilh hyolhyroidism in dogs. Vel Ialh 18:299-309.
23. Weelman AI 2000 Chronic auloimmune lhyroidilis. In: Werner & Ingbar's The Thyroid. 8
lh

edilion. raverman LI and Uliger RD (Ids.). IhiIadeIhia, IA: Liincoll WiIIiams &
WiIkins. :721-732.
24. MiIIer A, NeIson RW, Scoll-Moncrieff }C, el aI. 1992 SeriaI lhyroid hormone concenlralion in
heaIlhy eulhyroid dogs, dogs vilh hyolhyroidism, and eulhyroid dogs vilh aloic
dermalilis. r Vel } 148:451-457.
25. Nachreiner RI, RefsaI KR, Graham IA, ovman MM 2002 IrevaIence of serum lhyroid
hormone auloanlibodies in dogs vilh cIinicaI signs of hyolhryoidism. } Am Vel Med Ass
220:466-471.
26. Sauve I, Iaradis M 2000 Use of recombinanl human lhyroid-slimuIaling hormone for
lhyrolrohin slimuIalion lesl in eulhyroid dogs. Can Vel } 41:215-219.
27. Nachreiner RI, Iorsberg M, }ohnson CA, RefsaI KR 1995 VaIidalion of an assay for canine
TSH (cTSH). } Vel Inl Med 9:184.
28. Warlofsky L, urman KD, RingeI MD 1999 Trading one dangerous dogma for anolher`
Thyroid hormone lrealmenl of lhe eulhyroid sick syndrome. } CIin IndocrinoI Melab 84:1759-
1760.
29. Mei| I, MoI }A, evers MM, Ri|nberk A 1997 ResiduaI iluilary funclion afler
lransshenoidaI hyohyseclomy in dogs vilh iluilary-deendenl hyeradrenocorlicism. }
Indocr 155:531-539.
30. Larsen IR, Davies TI, SchIumberger M-}, Hay ID 2003 Thyroid hysioIogy and diagnoslic
evaIualion of alienls vilh lhyroid disorders. In: WiIIiams Texlbook of IndocrinoIogy, 10
lh


Chapter 3
_____________________________________________________________________________
68

edilion. Larsen IR, Kronenberg HM, MeImed S, IoIonsky KS (Ids). IhiIadeIhia, IA: W
Saunders. :331-373.
31. Saberi M, Uliger RD 1975 Augmenlalion of lhyrolroin resonses lo lhyrolroin-reIeasing
hormone foIIoving smaII decreases in serum lhyroid hormone concenlralions. } CIin
IndocrinoI Melab 40:435-441.
32. Scoll-Moncrieff }CR, NeIson RW 1998 Change in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolrohin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and dogs vilh concurrenl disease. } Am Vel Med Assoc 213:1435-
1438.
33. Docler R, Krenning II, de }ong M, Hennemann G 1993 The sick eulhyroid syndrome:
changes in lhyroid hormone serum aramelers and hormone melaboIism. CIin IndocrinoI
39:499-518.
34. Deeg C, Kasers A, Harlmann K, el aI. 1997 Canine Hyolhyreose: Nachveis von
Auloanlikrern gegen ThyreogIobuIin. |Canine hyolhyroidism: deleclion of anli-
lhyrogIobuIin auloanlibodies.j TierrzlI Irax 25:170-173.
35. Larsen IR, Davies TI 2003 Hyolhyroidism and lhyroidilis. In: WiIIiams Texlbook of
IndocrinoIogy, 10
lh
edilion Larsen IR, Kronenberg HM, MeImed S, IoIonsky KS (Ids).
IhiIadeIhia, IA: W Saunders. :423-455.
36. Nachreiner RI, RefsaI KR, Graham IA, el aI. 1998 IrevaIence of auloanlibodies lo
lhyrogIobuIin in dogs vilh nonlhyroidaI iIIness. Am } Vel Res 59:951-955.
37. Dixon RM, Mooney CT 1999 Canine serum lhyrogIobuIine auloanlibodies in heaIlh,
hyolhyroidism and non-lhyroidaI iIIness. Res Vel Sci 66:243-246.
38. }avadi S, D|a|adiningral-Laanen SC, Kooislra HS, el aI. 2005 Irimary hyeraIdosleronism, a
medialor of rogressive renaI disease. Domesl Anim IndocrinoI 28:85-104.
39. }avadi S, GaIac S, oer I, el aI. 2006 AIdoslerone lo renin ralio and corlisoI lo
adrenocorlicolrohic hormone ralio in heaIlhy dogs and in dogs vilh rimary
hyoadrenocorlicism. } Vel Inl Med 20:556-61.
40. Suciira MS, Camargo II, NickoIoff IL, el aI. 1983 The roIe of
99m
Tc erlechnelale ulake in
lhe evaIualion of lhyroid funclion. Inl } NucI Med ioI 10:29-33.
41. Na AMI, IoIIak YWIA, van den rom WI, Ri|nberk A 1994 Quanlilalive asecls of lhyroid
scinligrahy vilh erlechnelale (
99m
TcO4
-
) in cals. } Vel Inl Med 8:302-303.
42. Adams WH, DanieI G, Ielersen MG, Young K 1997 Quanlilalive
99m
Tc-erlechnelale lhyroid
scinligrahy in normaI beagIes. Vel RadioI UIlrasound 38: 323-328.





Chaptcr 4

5ccrctinn pattcrn nI thyrnid-stimu!ating hnrmnnc
in dngs during cuthyrnidism and hypnthyrnidism









H.5. Knnistra, M. Diaz-Espincira, J.A. Mn!,
W.E. van dcn Brnm and A. Ri|nbcrk

Dnmcstic Anima! Endncrinn!ngy 2000, 18: 19-29

_________________________________________________________

Dearlmenl of CIinicaI Sciences of Comanion AnimaIs
IacuIly of Velerinary Medicine, Ulrechl Universily, Ulrechl, The NelherIands


Chaler 4
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70
!'

In as many as one-lhird of dogs vilh rimary hyolhyroidism a Iasma
lhyrolroin (TSH) concenlralion vilhin lhe reference range for eulhyroid dogs is
found. To delermine vhelher lhis is due lo fIuclualions in lhe reIease of TSH, lhe
Iasma rofiIes of TSH vere anaIysed in 7 beagIe bilches by coIIecling bIood
samIes every 10 min for 6 hours, bolh before and afler induclion of rimary
hyolhyroidism.
Afler induclion of rimary hyolhyroidism, a 37-foId increase in mean
basaI Iasma TSH concenlralion and a 34-foId increase in mean area under lhe
curve for TSH vere found. AnaIysis by lhe IuIsar rogram demonslraled uIsaliIe
secrelion of TSH in lhe hyolhyroid slale, characlerized by reIaliveIy Iov
amIilude uIses (mean ( SIM) amIilude 41 3% of basaI Iasma TSH IeveI)
and a mean uIse frequency of 2.0 0.5 uIses/6h. In lhe eulhyroid slale,
significanl TSH uIses vere idenlified in onIy 2 dogs. The mean basaI Iasma TSH
IeveI correIaled osiliveIy (r0.84) vilh lhe mean amIilude of lhe TSH uIses, and
correIaled negaliveIy (r -0.88) vilh lhe TSH uIse frequency.
The resuIls of lhis sludy demonslrale uIsaliIe secrelion of TSH in dogs
during hyolhyroidism and onIy smaII fIuclualions in Iasma TSH concenlralions
during eulhyroidism. The findings aIso suggesl lhal lhe Iov TSH vaIues
occasionaIIy found in dogs vilh sonlaneous rimary hyolhyroidism may in
some cases in arl be lhe resuIl of uIlradian fIuclualions.










Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

71

Intrnductinn

Irimary hyolhyroidism is one of lhe mosl common endocrine diseases in
lhe dog. In lhe Iasl lvo decades, lhe diagnosis of rimary hyolhyroidism in dogs
has been based uon a Iov Iasma lhyroxine concenlralion lhal is nol resonsive
lo lhyrolroin (TSH) slimuIalion.
1
ecause TSH is no Ionger commerciaIIy
avaiIabIe for diagnoslic uroses in velerinary raclice, lhe diagnosis of canine
rimary hyolhyroidism is nov oflen based uon lhe combinalion of a Iov
lhyroxine concenlralion and an eIevaled TSH concenlralion vilhin a singIe Iasma
samIe.
2,3
Hovever, il has been reorled lhal in as many as one-lhird of dogs vilh
rimary hyolhyroidism lhe Iasma TSH concenlralion is vilhin lhe reference
range for eulhyroid dogs.
3-8
IossibIe reasons vhy dogs vilh rimary
hyolhyroidism have Iasma TSH concenlralions vilhin lhe reference range
incIude concurrenl secondary or lerliary hyolhyroidism,
5
faiIure of lhe TSH assay
lo delecl aII isoforms of circuIaling TSH,
3
and a decrease in eIevaled TSH vaIues in
hyolhyroid dogs vilh lime.
9
In addilion, fIuclualions in lhe circuIaling
concenlralions of TSH couId be a reason for lhis henomenon.
In humans and rals, frequenl samIing sludies have demonslraled lhal
TSH is secreled inlermillenlIy. In 1965 akke and Lavrence reorled circadian
TSH rhylhmicily in lhe iluilary gIand of eulhyroid and hyolhyroid rals.
10
Wilh
lhe deveIomenl of a sensilive radioimmunoassay for TSH, a circadian TSH
secrelion allern vas aIso reorled in humans.
11
Iarker .
12
vere lhe firsl lo
demonslrale a uIsaliIe or uIlradian TSH secrelion allern in humans. The
uIlradian TSH reIease in heaIlhy humans foIIovs a high-frequency, Iov-amIilude
uIsaliIe allern suerimosed on a Iov-frequency, high-amIilude circadian TSH
rhylhm.
13,14,15
UIlradian uIsaliIe TSH secrelion has aIso been demonslraled in
hyolhyroid bul nol in eulhyroid rals.
16

AII iluilary hormones sludied lhus far in lhe dog are aIso reIeased in a
uIsaliIe allern.
17-20
Hovever, lhere is aImosl no informalion on lhe TSH secrelion
allern in lhis secies. In a sludy by runer .
7
, serum samIes vere coIIecled al
2-hour inlervaIs and fIuclualions in TSH concenlralions vere delecled lhroughoul
lhe day in bolh eulhyroid and hyolhyroid dogs. AIlhough lhese aulhors vere
unabIe lo idenlify any diurnaI or uIsaliIe secrelion allern, lhe random
fIuclualions in serum TSH concenlralion may suggesl uIsaliIe secrelion of TSH in
lhe dog.
In lhis sludy ve anaIysed lhe 6-hour secrelion allerns of TSH in 7 beagIe
bilches, bolh before and afler induclion of rimary hyolhyroidism.

Chaler 4
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72
Matcria!s and Mcthnds

Ocgs
Seven heaIlhy, neulered beagIe bilches enlered lhe sludy al lhe age of 3
years. The dogs vere housed singIy in indoor-ouldoor runs, fed a slandard
commerciaI dry dog food lvice daiIy, and given valer a! |i|iiun. The daiIy food
inlake of lhe dogs vas aroximaleIy 300 grams. According lo lhe manufaclurer,
lhe iodine conlenl of lhe commerciaI dry dog food vas 250-300 g er 100 gram.
On lhe days on vhich bIood samIes vere coIIecled, lhe dogs vere fed lheir usuaI
commerciaI dog food belveen 09.00 and 09.30h.
Iulhyroidism vas confirmed by lhe finding of basaI and bovine TSH-
slimuIaled Iasma lhyroxine concenlralions vilhin lheir reseclive reference
ranges, a Iasma TSH concenlralion vilhin lhe reference range, a normaI image of
lhe lhyroid gIands oblained by scinligrahy using radioaclive erlechnelale
(
99m
TcO4
-
), and, evenluaIIy, by lhe finding of normaI lhyroid gIand lissue by
hisloIogicaI examinalion afler lhyroideclomy.
Irimary hyolhyroidism vas induced by surgicaI removaI of lhe lhyroid
gIands, Ieaving lhe exlernaI aralhyroid gIands in siiu. Tvo monlhs afler surgery,
scinligrahy vilh
99m
TcO4
-
vas used lo delecl remnanl lhyroid gIand lissue.
AroximaleIy 1 monlh Ialer, lhe dogs vere fed a Iov-iodine diel (meal and vhile
bread) for 3 veeks lo increase lhe iodine ulake by any remnanl lhyroid lissue,
vhich vas lhen deslroyed by adminislering 5 mCi ( 185 Mq) of sodium-
131
I-
iodide oraIIy.

Sanp|c cc||cciicn
Iood samIes (0.8 mI) for lhe delerminalion of lhe Iasma concenlralions
of TSH and lolaI lhyroxine vere coIIecled al 10-minule inlervaIs belveen 08.00
a.m. and 2.00 .m. This vas done 1.5 monlhs rior lo lhe surgicaI lhyroideclomy
and again one monlh afler lhe adminislralion of sodium-
131
I-iodide. Iood samIes
vere coIIecled by |uguIar veniunclure, immedialeIy Iaced in chiIIed, hearin-
coaled lubes, and cenlrifuged. IIasma vas slored al -20C unliI assayed.

Hcrncnc !cicrninaiicn
IIasma TSH concenlralions vere delermined by a homoIogous soIid-
hase, lvo-sile chemiIuminescenl enzyme immunomelric assay (ImmuIile
R
canine
TSH, Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, USA) according lo lhe
inslruclions of lhe manufaclurer. The inlra-assay coefficienls of varialion vere
5.0%, 4%, and 3.8% al TSH IeveIs of 0.20, 0.50, and 2.6 g/I, resecliveIy. The
inlerassay coefficienls of varialion vere 6.3% and 8.2% al TSH IeveIs of 0.16 and 2.8
g/I, resecliveIy. The Iovesl deleclabIe amounl of TSH vas 0.03 g/I. VaIues of
Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

73

TSH measured vilh lhe ImmuIile
R
canine TSH assay afler seriaI diIulion of lhree
canine samIes shoved good Iinearily lhroughoul ils caIibralion range. To
eslabIish lhe siking recovery, a 13 I aIiquol of lhree siking soIulions (4.3, 8.5,
and 18 g/I) vas siked inlo 247 I aIiquols of lhree differenl canine samIes (0.46,
2.8, and 5.2 g/I). The recovery ranged from 94 lo 104%. These resuIls confirm lhal
lhe ImmuIile
R
canine TSH assay yieIds an accurale measure of canine TSH
lhroughoul ils caIibralion range. Cross-reaclivily vilh ISH and LH vas negIigibIe.
The uer Iimil of lhe reference range for lhe Iasma TSH concenlralion in
eulhyroid dogs in our Iaboralory is 0.6 g/I.
IIasma lolaI lhyroxine concenlralions vere delermined by a homoIogous
soIid-hase, chemiIuminescenl enzyme immunoassay (ImmuIile
R
canine TolaI T4,
DIC, Los AngeIes, USA) according lo lhe inslruclions of lhe manufaclurer and
vaIidaled for lhe dog by runer ci a|.
7
The inlra-assay coefficienls of varialion vere
13.8% and 8.2% al lhyroxine IeveIs of 8 and 25 nmoI/I, resecliveIy. The Iovesl
deleclabIe amounl of lhyroxine vas 1.5 nmoI/I. VaIues of lolaI lhyroxine measured
vilh lhe ImmuIile
R
canine TolaI T4 assay afler seriaI diIulion of four canine samIes
shoved good Iinearily lhroughoul ils caIibralion range. To eslabIish lhe siking
recovery, a 50 I aIiquol of lhree siking soIulions (219, 412, and 798 nmoI/I) vas
siked inlo 950 I aIiquols of four differenl canine samIes (7, 14, 15, and 23
nmoI/I). The recovery ranged from 96 lo 114%. These resuIls confirm lhal lhe
ImmuIile
R
canine TolaI T4 assay yieIds an accurale measure of canine lolaI
lhyroxine lhroughoul ils caIibralion range.

Oaia ana|qsis
The secrelion allern of TSH vas anaIysed using lhe IuIsar rogram
deveIoed by Merriam and Wachler.
21
The rogram idenlifies secrelory eaks by
heighl and duralion from a smoolhed baseIine, using lhe assay SD as a scaIe faclor.
The cul-off aramelers G1-G5 of lhe IuIsar rogram vere sel al 5.78, 2.89, 1.84,
1.27, and 0.89 limes lhe inlra-assay SD as crileria for acceling eaks 1, 2, 3, 4, and
5 oinls vide, resecliveIy,
22
resuIling in a faIse-osilive error rale Iess lhan 5%.
The smoolhing lime, a vindov used lo caIcuIale a running mean vaIue, vas sel al
5 hours. The veighl assigned lo eaks vas 0.05. The A, , and C-vaIues of lhe
IuIsar rogram, used lo caIcuIale lhe variance of lhe assay, vere sel al A0.05,
3.52, and C0.71. The vaIues exlracled from lhe IuIsar anaIyses incIuded: lhe
overaII mean of lhe smoolhed baseIine, lhe area under lhe curve (AUC), lhe
amIilude of lhe uIses, and lhe number of uIses. The AUC vas caIcuIaled above
lhe zero IeveI.
IIasma concenlralions of lhyroxine and TSH beIov lhe deleclion Iimil of
lhe assay vere considered lo be 1.5 nmoI/I and 0.03 g/I, resecliveIy. Differences
in lhe AUC and basaI concenlralions vere evaIualed by Sludenl's i-lesl for reIaled
Chaler 4
_________________________________________________________________________________


74
samIes (lvo-laiIed). CorreIalions belveen basaI IeveIs and uIse characlerislics
vere anaIysed using lhe Iinear correIalion coefficienl. VaIues are exressed as
mean SIM or as mean and range. I<0.05 vas considered significanl.

Ethics nI cxpcrimcntatinn

The exerimenls in lhis sludy vere aroved by lhe IlhicaI Commillee of
lhe IacuIly of Velerinary Medicine, Ulrechl Universily.

Rcsu!ts

Tvo monlhs afler lhyroideclomy, scinligrahy vilh
99m
TcO4
-
demonslraled
lhal remnanl lhyroid gIand lissue vas resenl in lhe neck al lhe normaI lhyroid
Iocalion (5 dogs), under lhe base of lhe longue (2 dogs), and al lhe base of lhe hearl
(3 dogs).
The mean Iasma lhyroxine concenlralions vas 20.7 2.6 nmoI/I rior lo
lhe lhyroideclomy and 2.5 0.4 nmoI/I one monlh afler lhe adminislralion of
131
I.
The Iasma lhyroxine IeveIs in lhe eulhyroid and hyolhyroid slales did nol
overIa. AIlhough minor fIuclualions vere observed in lhe 6-h secrelion allerns
of lhyroxine, neilher in eulhyroidism nor in hyolhyroidism vas lhyroxine
secreled in a uIsaliIe fashion. In bolh silualions lhe mean basaI 6-h Iasma
lhyroxine concenlralion and lhe mean basaI 6-h Iasma TSH concenlralion vere
nol significanlIy correIaled.
Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

75



Figurc 1. The 6-hour secrelory rofiIes of TSH () and lhyroxine (T4) () in a 3-
year-oId eulhyroid beagIe bilch. Significanl uIses, caIcuIaled by lhe IuIsar
rogram, are indicaled by an aslerisk.

Chaler 4
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76


Figurc 2. The 6-hour secrelory rofiIes of TSH () and lhyroxine (T4) () in lvo 3-
year-oId beagIe bilches afler induclion of rimary hyolhyroidism. Significanl
uIses, caIcuIaled by lhe IuIsar rogram, are indicaled by an aslerisk.

Afler induclion of rimary hyolhyroidism a 37-foId increase in mean
basaI Iasma TSH concenlralion and a 34-foId increase in mean AUC for TSH vere
found. The basaI Iasma TSH concenlralions (mean 3.7 0.6 g/I) and lhe AUCs
for TSH (23.3 4.0 g/I6h) in lhe hyolhyroid slale vere significanlIy higher
(I0.001) lhan lhe basaI Iasma TSH concenlralions (mean 0.10 0.03 g/I) and lhe

Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

77

AUCs for TSH (mean 0.68 0.32 g/I6h) before induclion of rimary
hyolhyroidism.
AnaIysis of lhe TSH secrelory rofiIes in eulhyroidism (summarized in
TabIe 1) reveaIed lhe resence of significanl TSH uIses in onIy 2 of lhe 7 dogs,
aIlhough fIuclualions in lhe Iasma TSH concenlralion vere evidenl in 5 dogs
lhroughoul lhe 6-h eriod of 10-min samIing. An examIe of lhe TSH allern in
one of lhe eulhyroid dogs is resenled in Iigure 1. In 2 dogs lhe Iasma TSH
concenlralions in lhe eulhyroid slale vere conlinuousIy beIov lhe deleclion Iimil
of lhe TSH assay.
Afler induclion of rimary hyolhyroidism, significanl TSH uIses vere
idenlified by lhe IuIsar rogram in aII bul one of lhe dogs. Tvo examIes are
resenled in Iigure 2. The TSH secrelory allerns in individuaI dogs varied
considerabIy in uIse frequency and uIse amIilude, and in aII dogs fIuclualions
around lhe mean baseIine vere observed (TabIe 1, Iigure 2). The mean TSH uIse
frequency in hyolhyroidism vas aroximaleIy 2 uIses er 6 h. The TSH uIses
vere reIaliveIy Iov, having a mean amIilude of 41 3% of lhe basaI Iasma TSH
IeveI. There vas a significanl osilive correIalion (r 0.84, I<0.05) belveen lhe
mean basaI 6-h Iasma TSH concenlralion and lhe mean amIilude of lhe TSH
uIses. The mean basaI 6-h Iasma TSH concenlralion vas negaliveIy correIaled (r
-0.88, I<0.05) vilh lhe TSH uIse frequency.




The resuIls of lhis sludy demonslrale uIsaliIe secrelion of TSH in dogs during
hyolhyroidism. Iiluilary TSH secrelion is conlroIIed by lhe slimuIalory aclion of
hyolhaIamic TRH and inhibilion via cenlraI doaminergic and somaloslalinergic
mechanisms, as veII as by an inhibilory aclion of lhyroid hormones al lhe cenlraI,
hyolhaIamic, and iluilary IeveIs.
23,24
Sludies in humans have demonslraled lhal
conlinuous infusions vilh doamine and somaloslalin suress mean circuIaling
TSH IeveIs bul lhe uIsaliIe allern of TSH secrelion remains.
25
Thyroid hormones,
vilh lheir Iong melaboIic haIf-Iives are aIso unIikeIy lo acl as raid moduIalors of
TSH secrelion. Therefore, in humans, and robabIy aIso in dogs, TRH is mosl
IikeIy lo be lhe driving force for lhe uIsaliIe TSH secrelion allern.
23,25



Chaler 4
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78

ResuIls of IuIsar anaIysis: mean basaI Iasma lhyroxine concenlralion and
uIse characlerislics of lhe 6-hour secrelory rofiIes of TSH in 7 beagIe bilches
before and afler lhe induclion of rimary hyolhyroidism. The vaIues are
exressed as mean SIM (and range). The minimum and maximum TSH vaIues
are given as ercenlage change in reIalion lo lhe basaI TSH IeveI.



In conlrasl lo lhe findings during rimary hyolhyroidism, lhe resuIls of
lhis sludy faiIed lo demonslrale irrefulabIy lhal TSH is secreled in a uIsaliIe
fashion in eulhyroid dogs. AIlhough lhere vere fIuclualions in circuIaling TSH
IeveIs in 5 eulhyroid dogs, in onIy 2 vas significanl uIsaliIily delecled by lhe
IuIsar rogram. SimiIar observalions have been reorled in rals
16
and eves.
26
Hovever, in humans uIsaliIe secrelion of TSH has been reorled in bolh rimary
hyolhyroidism and eulhyroidism.
14,27
The difference belveen eulhyroid humans
and eulhyroid dogs vilh regard lo TSH uIsaliIily may be lhe reIaliveIy Iov
Iasma TSH IeveIs in eulhyroid dogs, vhich couId be reIaled lo lhe high iodine
conlenl of commerciaI dog food. The minimum daiIy iodine requiremenl of lhe
aduIl beagIe dog has been eslimaled lo be 140 g.
28
A! |i|iiun consumlion of
mosl commerciaIIy manufaclured dry dog foods rovides lhe average dog vilh a
daiIy iodine inlake of al Ieasl 500 g and some foods rovide considerabIy more.
29

In dogs receiving Iess lhan 140 g iodine er day enIarged and sarseIy granuIaled
iluilary TSH-secreling ceIIs vere found, suggesling high secrelory aclivily.
28
In
conlrasl, lhe high iodine conlenl of commerciaI dog food imIies a reIaliveIy Iov
Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

79

slimuIus for TSH secrelion. ConsequenlIy, lhe mean basaI TSH IeveIs are Iov in
eulhyroid dogs and lhis may rohibil lhe deleclion of significanl TSH uIses,
because of lhe osilive correIalion demonslraled in lhe resenl sludy and in
severaI sludies in humans
14,24,27,30,31
belveen lhe mean basaI Iasma TSH
concenlralion and lhe mean amIilude of lhe TSH uIses. In addilion, lhe
significanl TSH uIses delecled in lhe dogs during hyolhyroidism had a
reIaliveIy Iov amIilude comared lo lhe basaI Iasma TSH IeveIs. Therefore, lhe
amIiludes of ossibIe TSH uIses in lhe eulhyroid dogs may have been so Iov
lhal, due lo lhe variabiIily of lhe TSH assay in lhe range of eulhyroid TSH IeveIs,
TSH uIses couId nol be delecled by lhe IuIsar rogram.
Iood samIes for lhe delerminalion of lhe Iasma TSH concenlralion
vere coIIecled al 10-min inlervaIs for 6 hours. AIlhough lhis 6-h eriod vas
sufficienl lo demonslrale lhe uIsaliIe secrelion of TSH in hyolhyroid dogs, lhis 6-
h eriod did nol aIIov an accurale eslimalion of lhe TSH uIse frequency. In lhe
resenl sludy lhe mean TSH uIse frequency vas aroximaleIy 2 uIses er 6 h.
Hovever, lhe range of lhe TSH uIse frequency in lhe 6 dogs vas considerabIe,
indicaling lhal samIing for a Ionger lime eriod is required lo delermine
accuraleIy lhe TSH uIse frequency.
The minor fIuclualions observed in lhe 6-h Iasma rofiIes of lhyroxine in
hyolhyroid dogs have been reorled before.
32
Hovever, lhe fIuclualions lhal ve
observed vere nol significanlIy differenl from lhe inlra-assay varialion. In
addilion, lhe Iasma concenlralion of a hormone vilh a bioIogicaI haIf-Iife of aboul
a day, such as lhyroxine,
33
cannol be execled lo vary slrongIy.
In normaI reguIalion of lhe hyolhaIamic-iluilary-lhyroid axis, even
smaII decremenls in circuIaling lhyroid hormone concenlralion resuIl in an
increase in basaI TSH and augmenlalion of lhe TSH resonse lo TRH.
34

NeverlheIess, neilher during eulhyroidism nor during hyolhyroidism vere lhe
mean basaI Iasma TSH concenlralion and lhe mean Iasma lhyroxine IeveI
significanlIy correIaled. This suggesls lhal lhe sel-oinl for basaI TSH secrelion and
sensilivily lo Iasma lhyroid hormone concenlralions may vary from dog lo dog.
The roIe of lhyroid hormones in lhe generalion of uIsaliIe TSH secrelion is
robabIy nol greal, since in humans no ma|or infIuence of lhyroid hormones on
TSH uIse frequency has been observed.
14,22
The roIe of lhyroid hormones in
reguIaling uIsaliIe TSH secrelion in humans aears lo be reslricled lo
moduIalion of lhe TSH uIse amIilude.
22

Sludies in humans have demonslraled lhal ovarian sex sleroids and
androgens are nol crilicaIIy invoIved in lhe generalion of uIsaliIe TSH
secrelion.
15,35
Therefore, lhe resenl resuIls vilh neulered bilches may mosl IikeIy
aIy for inlacl dogs and neulered maIe dogs as veII. AII dogs vere 3 years of age,
vhich is an age al vhich rimary hyolhyroidism is common. ecause Greensan
Chaler 4
_________________________________________________________________________________


80
.
31
have reorled lhal TSH uIse frequency in men is nol age deendenl, our
observalions may resumabIy be exlraoIaled lo olher ages.
Desile lhe fIuclualions in lhe 6-h Iasma rofiIe of TSH during
hyolhyroidism, Iasma TSH IeveIs in eulhyroidism and hyolhyroidism did nol
overIa. This underIines lhe discriminalory over of lhe delerminalion of lhe TSH
concenlralion in a singIe Iasma samIe in dislinguishing lhese 2 subgrous.
Hovever, lhe dogs used in lhe resenl sludy vere heaIlhy beagIe dogs examined
bolh before and afler induclion of hyolhyroidism by a combinalion of surgicaI
lhyroideclomy and
131
I lrealmenl. SimiIar lo lhe resuIls in olher sludies,
7,36
lhis
modeI of rimary hyolhyroidism resuIled in consislenl increases in lhe Iasma
TSH concenlralion. These changes are in conlrasl lo lhe much more moderale
increases in Iasma TSH concenlralions observed in mosl dogs vilh sonlaneous
rimary hyolhyroidism.
2-8
Moreover, in as many as one-lhird of lhe dogs vilh
naluraIIy occurring rimary hyolhyroidism, Iasma TSH concenlralions have
been found lo be vilhin lhe reference range for eulhyroid dogs.
3-8
The reason for
lhis difference belveen exerimenlaI and sonlaneous rimary hyolhyroidism is
sliII unknovn bul il raises lhe queslion of vhelher dogs vilh exerimenlaIIy
induced rimary hyolhyroidism are an adequale modeI for naluraIIy occurring
canine rimary hyolhyroidism. NeverlheIess, lhe fIuclualions around lhe TSH
baseIine in exerimenlaIIy induced rimary hyolhyroid dogs in lhe resenl sludy
may in arl exIain vhy in some sonlaneous rimary hyolhyroid dogs a Iasma
TSH concenlralion vilhin lhe reference range for eulhyroid dogs is found. The
mean Iasma TSH concenlralion in dogs vilh sonlaneous rimary
hyolhyroidism has been reorled lo exceed lhe uer Iimil of lhe reference range
for TSH in eulhyroid dogs onIy marginaIIy.
3,6,7
ConsequenlIy, fIuclualions around
lhe TSH baseIine al lhis IeveI may occasionaIIy resuIl in Iasma TSH
concenlralions vilhin lhe reference range for eulhyroid dogs. This Iine of
reasoning is suorled by lhe findings of runer
7
vho reorled lhal dogs
vilh sonlaneous rimary hyolhyroidism may have random fIuclualions in
Iasma TSH concenlralions, vilh some of lhe vaIues being oulside lhe reference
range for eulhyroid dogs and some vilhin.
In summary, lhe resuIls of lhis sludy demonslrale uIsaliIe secrelion of
TSH in dogs during hyolhyroidism and onIy smaII fIuclualions in Iasma TSH
concenlralion during eulhyroidism. The uIsaliIe secrelion allern of TSH vas
characlerized by a uIse frequency of aroximaleIy 2 uIses/6h and TSH uIses
vilh a reIaliveIy Iov amIilude comared vilh basaI IeveIs. The resuIls of lhis
sludy aIso suggesl lhal lhe Iov TSH vaIues occasionaIIy found in dogs vilh
sonlaneous rimary hyolhyroidism may in some cases in arl be lhe resuIl of
uIlradian fIuclualions.

Secrelion allern of lhyroid-slimuIaling hormone
_________________________________________________________________________________

81

Acknnw!cdgmcnts

The aulhors are gralefuI for lhe lechnicaI assislance of Ms. M.I. Ieelers,
Ms. A.M.T.C. van Wees, Mrs. }. WoIfsvinkeI, Ms. Y.W.I.A. IoIIak, Mr. H.G.H. van
IngeIen, Mr. M.}. IeI, and Mr. I.M.G. Senders. The crilicaI reading of lhe
manuscril by Dr. .I. eIshav is highIy arecialed. The assays used for lhe
delerminalion of lhe Iasma concenlralions of TSH vere a generous gifl from
Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, USA.







Chaler 4
_________________________________________________________________________________


82
%

1. eIshav I and Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Am Anim Hos Assoc 15:17-23.
2. Dixon RM, Graham IA and Mooney CT 1996 Serum lhyrolroin concenlralions: a nev
diagnoslic lesl for canine hyolhyroidism. Vel Rec 138:594-595.
3. Ielerson MI, MeIian C and NichoIs R 1997 Measuremenl of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroin concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Assoc 211:1396-1402.
4. }ensen AL, Iversen L, Hoier R, Krislensen I and Henriksen I 1996 IvaIualion of an
immunoradiomelric assay for lhyrolroin in serum and Iasma samIes of dogs vilh
rimary hyolhyroidism. } Com IalhoI 114:339-346.
5. Ramsey IK, Ivans H and Herrlage MI 1997 Thyroid-slimuIaling hormone and lolaI lhyroxine
concenlralions in eulhyroid, sick eulhyroid, and hyolhyroid dogs. } SmaII Anim Iracl
38:540-545.
6. Scoll-Moncrieff }CR, NeIson RW, runer }M and WiIIiams DA 1998 Comarison of serum
concenlralions of lhyroid-slimuIaling hormone in heaIlhy dogs, hyolhyroid dogs, and
eulhyroid dogs vilh concurrenl disease. } Am Vel Med Assoc 212:387-391.
7. runer }M, Scoll-Moncrieff }CR and WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am
Vel Med Asso 212:1572-1575.
8. Dixon RM and Mooney CT 1999 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40:72-78.
9. Nachreiner RI, Iorsberg M, }ohnson CA and RefsaI KR 1995 VaIidalion of an assay for canine
TSH (cTSH). } Vel Inl Med 9:184.
10. akke }L and Lavrence N 1965 Circadian eriodicily in lhyroid slimuIaling hormone lilre in
lhe ral hyohysis and serum. MelaboIism 14:841-846.
11. VanhaeIsl L, van Cauler I, Degaule }I and GoIslein } 1972 Circadian varialions of serum
lhyrolroin IeveIs in man. } CIin IndocrinoI Melab 35:479-482.
12. Iarker DC, Iekary AI and Hershman }M 1976 Iffecl of normaI and reversed sIee-vake
uon nyclohemeraI rhylhmicily of Iasma lhyrolroin: evidence suggeslive of an inhibilory
infIuence of sIee. } CIin IndocrinoI Melab 43:318-329.
13. rabanl G, Ranfl U, Ocran K, Hesch RD and von zur MhIen A 1986 Thyrolroin-an
eisodicaIIy secreled hormone. Acla IndocrinoI 112:315-322.
14. Greensan SL, KIibanski A, SchoenfeId D and Ridgvay IC 1986 IuIsaliIe secrelion of
lhyrolroin in man. } CIin IndocrinoI Melab 63:661-668.
15. rabanl G, Irank K, Ranfl U, Schuermeyer Th, Wagner TOI, Hauser H, Kummer , Ieislner
H, Hesch RD and von zur MhIen A 1990 IhysioIogicaI reguIalion of circadian and uIsaliIe
lhyrolroin secrelion in normaI man and voman. } CIin IndocrinoI Melab 70:403-409.

Secrelion allern of lhyroid-slimuIaling hormone
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83

16. ruhn TO, McIarIane M, Deckey }I and }ackson IMD 1992 AnaIysis of uIsaliIe secrelion of
lhyrolroin and grovlh hormone in lhe hyolhyroid ral. IndocrinoIogy 131:2615-2621.
17. Kemainen R} and Sarlin }L 1984 Ividence for eisodic bul nol circadian aclivily in Iasma
concenlralions of adrenocorlicolrohin, corlisoI and lhyroxine in dogs. } IndocrinoI 103:219-
226.
18. Irench M, Vialkus I, Cukerman I, Sirek A and Sirek OV 1987 Secrelory allern of canine
grovlh hormone. Am } IhysioI 252:I268-I272.
19. Kooislra HS, Greven SH, MoI }A and Ri|nberk A 1997 IuIsaliIe secrelion of -MSH and lhe
differenliaI effecls of dexamelhasone and haIoeridoI on lhe secrelion of -MSH and ACTH
in dogs. } IndocrinoI 152:113-121.
20. Kooislra HS, Okkens AC, evers MM, Io-Sni|ders C, van Haaflen , DieIeman S} and
Schoemaker } 1999 Concurrenl uIsaliIe secrelion of Iuleinizing hormone and foIIicIe-
slimuIaling hormone during differenl hases of lhe eslrous cycIe and aneslrus in beagIe
bilches. ioI Rerod 60:65-71.
21. Merriam GR and Wachler KW 1982 AIgorilhms for lhe sludy of eisodic hormone secrelion.
Am } IhysioI 243:I310-I318.
22. rabanl G, rabanl A, Ranfl U, Ocran K, KhrIe }, Hesch RD and von zur MhIen A 1987
Circadian and uIsaliIe lhyrolroin secrelion in eulhyroid man under lhe infIuence of lhyroid
hormone and gIucocorlicoid adminislralion. } CIin IndocrinoI Melab 65:83-88.
23. ehrends }, Irank K, Dogu I and rabanl G 1998 CenlraI nervous syslem conlroI of
lhyrolroin secrelion during sIee and vakefuIness. Horm Res 49:173-177.
24. rabanl G 1998 IuIsaliIe und zirkadiane TSH-Sekrelion. KIinische ReIevanz` Inlernisl 39:619-
622.
25. rabanl G, Irank K, Hoang-Vu C, Hesch RD and von zur MhIen A 1991 HyolhaIamic
reguIalion of uIsaliIe lhyrolroin secrelion. } CIin IndocrinoI Melab 72:145-150.
26. DahI GI, Ivans NI, Thrun LA and Karsch I} 1994 A cenlraI negalive feedback aclion of
lhyroid hormones on lhyrolroin-reIeasing hormone secrelion. IndocrinoIogy 135:2392-2397.
27. Adriaanse R, rabanl G, Irank K, Inderl I and Wiersinga WM 1992 Circadian changes in
uIsaliIe TSH reIease in rimary hyolhyroidism. CIin IndocrinoI 37:504-510.
28. eIshav I, Cooer T and ecker DV 1975 The iodine requiremenl and infIuence of iodine
inlake on iodine melaboIism and lhyroid funclion in lhe aduIl beagIe. IndocrinoIogy
96:1280-1291.
29. eIshav I 1983 Thyroid diseases. In: Texlbook of Velerinary InlernaI Medicine, Diseases of
lhe Dog and Cal. 2
nd
edilion. Illinger S} (ed.). IhiIadeIhia: W Saunders Co. 1592-1614.
30. Romi|n }A, Adriaanse R, rabanl G, Irank K, Inderl I and Wiersinga WM 1990 IuIsaliIe
secrelion of lhyrolroin during fasling: a decrease of lhyrolroin uIse amIilude. } CIin
IndocrinoI Melab 70:1631-1636.
31. Greensan SL, KIibanski A, Rove }W and IIahi D 1991 Age-reIaled aIleralions in uIsaliIe
secrelion of TSH: roIe of doaminergic reguIalion. Am } IhysioI 260:I486-I491.
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84
32. MiIIer A, NeIson RW, Scoll-Moncrieff }C, NeaI L and olloms GD 1992 SeriaI lhyroid
hormone concenlralions in heaIlhy eulhyroid dogs, dogs vilh hyolhyroidism, and
eulhyroid dogs vilh aloic dermalilis. r Vel } 148:451-458.
33. eIshav I, arandes M, ecker DV and erman M 1974 A modeI of iodine kinelics in lhe
dog. IndocrinoIogy 95:1078-1093.
34. Saberi M and Uliger RD 1974 Augmenlalion of lhyrolroin resonses lo lhyrolroin reIeasing
hormone foIIoving smaII decreases in serum lhyroid hormone concenlralions. } CIin
IndocrinoI Melab 40:435-441.
35. Rossmanilh WG, SlbIer C, enz R, ornslein SR and Scherbaum WA 1992 RoIe of ovarian
sex sleroids in lhe reguIalion of lhyrolroin (TSH) secrelion of hyogonadaI vomen. Acla
IndocrinoI 127:131-137.
36. WiIIiams DA, Scoll-Moncrieff }C, runer }, Suslarsic D, Ianosian-Sahakian N, Unver I and II
Shami AS 1996 VaIidalion of an immunoassay for canine lhyroid-slimuIaling hormone and
changes in serum concenlralion foIIoving induclion of hyolhyroidism in dogs. } Am Vel
Med Assoc 209:1730-1732.

Chaptcr 5


Primary hypnthyrnidism in dngs is assnciatcd
with c!cvatcd grnwth hnrmnnc rc!casc









W. M. Lcc, M. Diaz-Espincira, J. A. Mn!, A. Ri|nbcrk and H. 5. Knnistra


Jnurna! nI Endncrinn!ngy 2001, 168: 59-66

_________________________________________________________________________

Dearlmenl of CIinicaI Sciences of Comanion AnimaIs,
IacuIly of Velerinary Medicine, Ulrechl Universily, Ulrechl, The NelherIands

Chaler 5
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86

!$

The uIsaliIe secrelion allerns of grovlh hormone (GH) vere
invesligaled in seven beagIe bilches by coIIecling bIood samIes every 10 min for 6
h during eulhyroidism and 1.5 years afler induclion of rimary hyolhyroidism.
Hyolhyroidism vas induced by surgicaI removaI of lhe lhyroid gIands and
subsequenl deslruclion of any remnanl lhyroid lissue by oraI adminislralion of
sodium |
131
Ijiodide.
Some of lhe hysicaI changes observed in lhe dogs vilh rimary
hyolhyroidism mimicked lhose of acromegaIy. During bolh eulhyroidism and
hyolhyroidism GH vas secreled in a uIsaliIe fashion. The mean ( SIM) basaI
Iasma GH concenlralion vas significanlIy higher (I0.003) in lhe hyolhyroid
slale (4.1 1.6 g/I) lhan in lhe eulhyroid slale (1.2 0.4 g/I). Likevise, lhe mean
AUC for GH above lhe zero-IeveI during hyolhyroidism (27.0 10.0 g/Iiler6h)
vas significanlIy higher (I0.004) lhan lhal during eulhyroidism (11.7 2.0
g/I6h). The mean AUC for GH above lhe baseIine vas significanlIy Iover
(I0.008) during hyolhyroidism (2.4 0.8 g/I 6h) lhan during eulhyroidism (4.5
1.8 g/I 6h), vhereas lhere vas no significanl difference in GH uIse frequency.
The mean Iasma insuIin-Iike grovlh faclor (IGI)-I IeveI vas significanlIy higher
(I<0.01) in lhe hyolhyroid slale (169 45 g/I) lhan in lhe eulhyroid slale (97 15
g/I).
The resuIls of lhis sludy demonslrale lhal rimary hyolhyroidism in dogs
is associaled vilh eIevaled basaI GH secrelion and Iess GH secreled in uIses. This
eIevaled GH secrelion has endocrine significance as iIIuslraled by eIevaled Iasma
IGI-I IeveIs and some hysicaI changes mimicking acromegaIy. Il is discussed lhal
lhe increased GH reIease in hyolhyroid dogs may be lhe resuIl of lhe absence of a
resonse eIemenl for lhyroid hormone vilhin lhe canine iluilary GH gene and
aIleralions in sura-iluilary reguIalion.

IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

87

Introduction

Grovlh hormone (GH) is secreled by lhe iluilary anlerior Iobe in a
uIsaliIe fashion under lhe reguIalion of lvo hyolhaIamic elides: GH-reIeasing
hormone (GHRH) slimuIales GH secrelion, vhiIe somaloslalin (SS) inhibils GH
reIease. This reguIalory syslem is infIuenced by negalive feedback from
eriheraIIy formed grovlh faclors, arlicuIarIy insuIin-Iike grovlh faclor-1 (IGI-
1).
1,2
In addilion, a recenlIy idenlified GH-reIeasing elide, caIIed GhreIin, is IikeIy
lo Iay a roIe in lhe reguIalion of iluilary GH secrelion.
3
The amIilude and
frequency of GH secrelory uIses are reguIaled by a comIex array of exlernaI and
inlernaI slimuIi incIuding age,
4,5
gender,
6
eslrous cycIe hase,
7
genelic background,
8

nulrilionaI slalus,
9
sIee,
10
disease slalus,
11
and body comosilion.
12
In addilion,
hormones such as gIucocorlicoids and lhyroid hormones infIuence lhe uIsaliIe
secrelion allern of GH.
13

In rals, lhyroid hormones are necessary lo mainlain bolh uIsaliIe and
induced GH secrelion.
14
The GH secrelory rofiIe has been found lo shifl from a
Iov-frequency, high-amIilude allern in eulhyroid rals lo a high-frequency, Iov-
amIilude allern in rals vilh miId hyolhyroidism induced by melhimazoIe.
15

Severe hyolhyroidism in rals is associaled vilh markedIy reduced circuIaling
GH IeveIs,
16-20
aboIished GH bursls,
21,22
bIunled GH resonse lo GHRH,
18,23
and
decreased iluilary conlenl of mRNA encoding for GH.
18,24,25
In addilion,
hyolhyroidism in rals causes a reduclion in iluilary GH conlenl,
16,22,25,26,27

iluilary GHRH recelor mRNA,
18,28
hyolhaIamic SS conlenl,
18
and hyolhaIamic
SS mRNA.
18

In humans vilh rimary hyolhyroidism, lhe eak GH vaIues afler
GHRH adminislralion are Iov.
23,29,30,31
Severe rimary hyolhyroidism in chiIdren
is associaled vilh allenualed sonlaneous GH secrelion.
32,33,34
In Iine vilh lhese
observalions are lhe findings of in vilro sludies in ceII Iines derived from human
GH-secreling iluilary lumors vhich reveaIed lhal lhyroid hormones romole GH
mRNA accumuIalion
35
and GH reIease.
36
The slimuIalory effecl of lhyroid
hormones on GH mRNA accumuIalion has been ascribed lo lranscrilionaI
aclivalion of lhe human GH gene.
35

Informalion vilh regard lo changes in iluilary GH secrelion during
rimary hyolhyroidism in olher mammaIs is scarce. Lov circuIaling GH IeveIs
have been reorled in hyolhyroid igs.
37
In conlrasl, hyolhyroidism did nol
affecl sonlaneous GH secrelion in shee and sleers, aIlhough lhe resonse lo
slimuIalion vilh GHRH vas reduced in hyolhyroid shee.
38,39

In dogs, rimary hyolhyroidism is one of lhe mosl common endocrine
diseases. The signs and symloms incIude veighl gain, Ielhargy and
dermaloIogicaI changes such as lruncaI aIoecia and skin lhickening due lo
Chaler 5
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88

mucooIysaccharide accumuIalion.
40,41
Changes in lhe GH secrelion allern mighl
conlribule lo lhe fealures observed in dogs vilh rimary hyolhyroidism bul
informalion on lhe uIsaliIe GH secrelion allern in dogs vilh rimary
hyolhyroidism is Iacking. Therefore, ve invesligaled lhe 6-h uIsaliIe GH
secrelion allern bolh before and afler induclion of hyolhyroidism in seven
beagIe dogs.

Matcria!s and Mcthnds

Ocgs
Seven neulered (ovarieclomized) beagIe bilches enlered lhe sludy al lhe
age of 3 years. AII bilches vere accuslomed lo lhe Iaboralory environmenl and
rocedures such as lhe coIIeclion of bIood. They vere housed in individuaI cages,
fed a commerciaI dog food lvice a day, and had free access lo valer. On lhe days
on vhich bIood samIes vere coIIecled, lhe dogs vere fed lheir usuaI commerciaI
dog food belveen 09:00 and 09:30 h.
Iulhyroidism vas confirmed by lhe finding of basaI and bovine
lhyrolroin (TSH)-slimuIaled Iasma lhyroxine concenlralions vilhin lheir
reseclive reference range, a Iasma TSH concenlralion vilhin lhe reference range,
a normaI image of lhe lhyroid gIands oblained by scinligrahy using radioaclive
erlechnelale (
99m
TcO4
-
), and, evenluaIIy, by lhe finding of normaI lhyroid gIand
lissue by hisloIogicaI examinalion afler lhyroideclomy.
Irimary hyolhyroidism vas induced by surgicaI removaI of lhe lhyroid
gIands, Ieaving lhe exlernaI aralhyroid gIands in silu. Aboul 3 monlhs Ialer, lhe
dogs vere fed a Iov-iodine diel (meal and vhile bread) for 3 veeks lo increase lhe
iodine ulake by any remnanl lhyroid lissue, vhich vas lhen deslroyed by
adminislering 5 mCi (185 Mq ) of sodium |
131
Ijiodide oraIIy.

Sanp|c cc||cciicn
Iood samIes for delermining lhe Iasma concenlralion of GH vere
coIIecled al 10-min inlervaIs belveen 08.00 and 14.00h. This vas done one monlh
rior lo lhe surgicaI lhyroideclomy and again 1.5 years Ialer. Iood samIes vere
coIIecled by |uguIar veneunclure, vhich vere lhen immedialeIy Iaced in chiIIed
IDTA-coaled lubes, and cenlrifuged. IIasma vas slored al 20 C unliI assayed.

Hcrncnc !cicrninaiicn
IIasma TSH concenlralions vere delermined by a homoIogous soIid-
hase, lvo-sile chemiIuminescenl enzyme immunomelric assay (ImmuIile canine
TSH, Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, CA) in accordance
vilh lhe inslruclions of lhe manufaclurer. The inlra-assay coefficienls of varialion
IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

89

vere 5.0, 4.0 and 3.8% al TSH IeveIs of 0.20, 0.50, and 2.6 g/I, resecliveIy. The
inlra-assay coefficienls of varialion vere 6.3 and 8.2% al TSH IeveIs of 0.16 and 2.8
g/I, resecliveIy. The Iovesl deleclabIe amounl of TSH vas 0.03 g/I.
IIasma lolaI lhyroxine concenlralions vere delermined by a homoIogous
soIid-hase, chemiIuminescenl enzyme immunoassay (ImmuIile canine TolaI T4,
DIC, Los AngeIes, CA) in accordance vilh lhe inslruclions of lhe manufaclurer
and vaIidaled for lhe dog by runer ci a|.
42
The inlra-assay coefficienls of varialion
vere 13.8 and 8.2 % al lhyroxine IeveIs of 8 and 25 nmoI/I, resecliveIy. The Iovesl
deleclabIe amounl of lhyroxine vas 1.5 nmoI/I.
IIasma GH concenlralions vere measured by a homoIogous
radioimmunoassay as described reviousIy.
43
The inlra- and inler-assay
coefficienls of varialion vere 3.8 and 7.2%, resecliveIy, and lhe sensilivily of lhe
assay vas 0.3 g/I.
IIasma IGI-I concenlralions vere measured by a heleroIogous RIA as
described reviousIy.
44
The inlra- and inler-assay coefficienls of varialion vere 4.7
and 15.6%, resecliveIy, al an IGI-1 concenlralion of 175 g/I. The sensilivily of lhe
assay vas 6 g/I.

Oaia ana|qsis
The 6-hour secrelion allerns of GH vere anaIyzed using lhe IuIsar
rogram deveIoed by Merriam and Wachler.
45
The rogram idenlifies secrelory
eaks by heighl and duralion from a smoolhed baseIine, using lhe assay SD as a
scaIe faclor. The cul-off aramelers G1-G5 of lhe IuIsar rogram vere sel al 3.98,
2.4, 1.68, 1.24, and 0.93 limes lhe assay SD as crileria for acceling eaks 1, 2, 3, 4,
and 5 oinls vide, resecliveIy, resuIling in a faIse-osilive error rale of Iess lhan
5%. The smoolhing lime, a vindov used lo caIcuIale a running mean vaIue, vas
sel al 5 h. The veighl assigned lo eaks vas 0.05. The A, , and C vaIues of lhe
IuIsar rogram, used lo caIcuIale lhe variance of lhe assay, vere sel al A0, 7.2,
and C5. The vaIues exlracled from lhe IuIsar anaIyses incIuded lhe overaII mean
of lhe smoolhed baseIine, lhe number of eaks, lhe area under lhe curve (AUC)
above lhe zero IeveI, and lhe AUC above lhe baseIine.
Differences in aramelers during eulhyroidism and hyolhyroidism vere
evaIualed by Sludenl's i-lesl for reIaled samIes (lvo-laiIed). Since lhe dala vere
nol assumed lo be normaIIy dislribuled, differences in uIse frequency vere
delermined by non-aramelric anaIysis, using WiIcoxon's signed rank lesl. VaIues
are exressed as mean SIM or as mean and range. I<0.05 vas considered
significanl.

Chaler 5
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90

Ethics nI cxpcrimcntatinn

The exerimenls in lhis sludy vere aroved by lhe IlhicaI Commillee of
lhe IacuIly of Velerinary Medicine, Ulrechl Universily.

Rcsu!ts

The mean body veighl during hyolhyroidism (20.1 1.4 kg) vas
significanlIy higher (I0.002) lhan lhal during eulhyroidism (14.5 0.7 kg).
Induclion of rimary hyolhyroidism aIso resuIled in marked exercise inloIerance
and skin changes such as lhick foIding. In aII dogs, aIoecia and
hyerigmenlalion vere noliceabIe eseciaIIy on lhe bridge of lhe nose. In four of
lhe dogs lhere aeared lo be some maxiIIar rognalhia vilh videning of lhe
inlerdenlaI saces.
Al 1.5 years afler induclion of rimary hyolhyroidism Iasma lhyroxine
concenlralions vere Iov (< 2 nmoI/I) as comared lo lhe eulhyroid slale (18.3 1.7
nmoI, I<0.001). The mean Iasma TSH IeveI rose (I0.01) from 0.11 0.03 g/I
rior lo lhyroideclomy lo 1.4 0.38 g/I in hyolhyroidism.
GH vas secreled in a uIsaliIe fashion bolh before and afler induclion of
rimary hyolhyroidism. IxamIes of lhe uIsaliIe secrelion allerns of GH
during eulhyroidism and hyolhyroidism are shovn in Iigure 1. The mean basaI
Iasma GH IeveI in lhe hyolhyroid slale (4.1 1.6 g/I) vas significanlIy higher
(I0.003) lhan lhal during eulhyroidism (1.2 0.4 g/I). Likevise, lhe mean AUC
for GH above lhe zero-IeveI (27.0 10.0 g/I 6h) vas significanlIy higher (I0.004)
lhan lhal during eulhyroidism (11.7 2.0 g/I 6h). In conlrasl, lhe AUC for GH
above lhe baseIine vas significanlIy Iover (I0.008) during hyolhyroidism (2.4
0.8 g/I 6h) lhan during eulhyroidism (4.5 1.8 g/I 6h). The GH uIse frequency
in lhe 6 h secrelory allerns varied from one lo len eaks rior lo lhyroideclomy,
and from lvo lo five eaks afler induclion of hyolhyroidism (TabIe 1). The mean
GH uIse frequency in lhe eulhyroid slale (6 3 uIses/6h) vas higher lhan lhal in
lhe hyolhyroid slale (4 1 uIses/6h), bul lhis difference vas nol slalislicaIIy
significanl (I0.24). The mean Iasma IGI-1 IeveI vas significanlIy higher
(I0.009) in lhe hyolhyroid slale (169 16 g/I) lhan in lhe eulhyroid (97 15
g/I).


IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

91



5$ - Characlerislics of lhe 6 h secrelory rofiIes of GH and lhe Iasma IGI-1
IeveIs in seven bilches during eulhyroidism and hyolhyroidism. asaI GH, basaI
Iasma GH IeveI, AUC(0), AUC above zero-IeveI, AUC (basaIIine), AUC above
baseIine, frequency, GH uIse frequency er 6 h.



Chaler 5
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92



Figurc 1. The 6-hr secrelory rofiIes of GH in a beagIe bilch during eulhyroidism
(before) and during hyolhyroidism (afler). Significanl GH uIses, caIcuIaled by
lhe IuIsar rogram, are indicaled by an aslerisk.

IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

93

)

The resuIls of lhis sludy demonslrale lhal in dogs rimary hyolhyroidism
is associaled vilh eIevaled GH secrelion. In conlrasl lo observalions in olher
mammaIs, in dogs bolh basaI and inlegraled GH IeveIs vere significanlIy higher in
hyolhyroidism lhan in eulhyroidism. In some mammaIs, such as shee and
sleers, rimary hyolhyroidism does nol seem lo affecl GH secrelion,
38,39
vhereas
in rals
16-20
and igs
37
rimary hyolhyroidism is associaled vilh Iov circuIaling GH
IeveIs.
The human iluilary GH gene conlains a osilive lhyroid hormone
resonse eIemenl (TRI).
35
In Iine vilh lhis observalion, exerimenlaIIy induced
lhyroid hormone excess Ieads lo increased GH secrelion in heaIlhy men.
46
Il is
IikeIy lhal lhe Iov lhyroid hormone IeveIs of hyolhyroidism viII resuIl in a Iov
exression of iluilary GH in secies vilh a osilive TRI in lhe GH gene. Indeed,
in hyolhyroid rals bolh decreased iluilary conlenl of GH mRNA
18,24,25
and
reduced iluilary GH conlenl
16,22,25,26,27
have been reorled. The Iack of a decrease in
GH biosynlhesis and reIease in dogs vilh rimary hyolhyroidism may be
exIained by lhe absence of a osilive TRI vilhin lhe canine GH gene.
AIlernaliveIy, lhe resence of a negalive TRI vilhin lhe GH gene vouId resuIl in a
reduced suression of GH exression, i.e. eIevaled GH roduclion and secrelion
in hyolhyroid animaIs. The resence of negalive TRIs has been demonslraled in
differenl gene, for examIe in lhe TSH-reIeasing hormone (TRH) gene,
47
lhe
TSHbela gene,
48
and in keralin genes.
49
Sequence anaIysis of lhe romoler region
and inlron 1 of lhe canine GH gene
50
has nol demonslraled lhe exislence of a TRI
in lhis secies. Hovever, geI shifl assays or foolrinls have nol been erformed yel
in lhe dog.
Aarl from direcl effecls in lranscrilion of lhe GH gene, il is ossibIe lhal
hyolhyroidism affecls GH reIease via an infIuence on lhe hyolhaIamic hormones
GHRH and SS. In rals, rimary hyolhyroidism is associaled vilh a reduced
hyolhaIamic conlenl of SS mRNA and SS.
18
Moreover, lhyroideclomy in rals Ieads
lo decreased SS reIease from lhe hyolhaIamus.
19
Therefore, il may be osluIaled
lhal lhe hyolhaIamic SS secrelion is aIso reduced in dogs vilh rimary
hyolhyroidism, vhich may in lurn exIain lhe eIevaled basaI GH secrelion in lhis
secies.
Anolher olion lhal shouId be discussed concerns lhe feedback signaI of
lhe Iov lhyroxine concenlralion on lhe reIease of hyolhaIamic TRH. In heaIlhy
humans adminislralion of TRH does nol resuIl in areciabIe changes in Iasma
GH IeveIs,
51,52,53
vhereas in eulhyroid rals slimuIalion vilh TRH eIicils no
16
or onIy
a smaII lransienl rise in Iasma GH IeveIs.
26
In conlrasl, in humans and rals vilh
rimary hyolhyroidism TRH adminislralion resuIls in significanlIy increased
Chaler 5
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94

Iasma GH IeveIs.
16,51,52,53
In Iine vilh lhese observalions, Konaka .
54
have
demonslraled lhe exression of TRH-recelor mRNA in ral somalolroes. The
aradoxicaI resonse of GH lo TRH in rimary hyolhyroidism may be caused by
lhe reIalive deficiency of SS discussed in lhe revious aragrah, as SS infusion
inhibils lhe GH resonse lo TRH in hyolhyroid human alienls.
55
AIso, in heaIlhy
dogs TRH adminislralion does nol resuIl in changes in lhe Iasma GH IeveI.
56

Hovever, so far lhere is no informalion on lhe effecl of TRH on GH secrelion in
dogs vilh rimary hyolhyroidism. NeverlheIess, il may be hyolhesized lhal aIso
in hyolhyroid dogs lhe eIevaled reIease of TRH nol onIy causes increased TSH
secrelion bul may aIso give rise lo eIevaled GH secrelion.
In agreemenl vilh revious ubIicalions,
57,58,59
in lhe resenl dogs GH vas
secreled in a uIsaliIe fashion. The resuIls demonslrale lhal rimary
hyolhyroidism in dogs is associaled vilh changes in lhe uIsaliIe secrelion
allern of GH. The mean AUC for GH above lhe baseIine vas significanlIy Iover
during hyolhyroidism lhan in lhe eulhyroid slale. In combinalion vilh an
unchanged GH uIse frequency lhis indicales lhal in hyolhyroid dogs Iess GH is
reIeased er uIse. This may be exIained by a bIunled resonse of lhe iluilary
somalolroes lo GHRH. Indeed, adminislralion of GHRH resuIls in a markedIy
reduced GH resonse in hyolhyroid humans,
23,29,30,31
hyolhyroid rals,
18,23
and
hyolhyroid shee.
39
Moreover, in hyolhyroid rals a reduced iluilary conlenl of
GHRH recelor mRNA has been reorled.
18,28

The reduclion in GH secreled in uIses in lhe hyolhyroid dogs may aIso
be exIained by increased IGI-I secrelion. The resuIls demonslrale lhal lhe Iasma
IGI-I IeveIs vere significanlIy higher in hyolhyroidism lhan in eulhyroidism.
IIevaled circuIaling IeveIs of IGI-I have been reorled lo inhibil uIsaliIe iluilary
GH secrelion,
1
mainIy by allenualion of sonlaneous GH uIse amIilude.
2

IinaIIy, lhe reduclion in GH secreled in uIses in hyolhyroid dogs may
be ascribed lo exlra-iluilary secrelion of GH. Decreased GH uIsaliIily has been
reorled in vomen during lhe second haIf of regnancy.
60
In lhese vomen lhe Ioss
of GH uIsaliIily is due lo lhe reIease of a IacenlaI GH varianl.
60
In addilion, a
reduclion in GH secreled in uIses has been reorled in bilches during lhe
rogeslerone hase of lhe eslrous cycIe
59
and in bilches lrealed vilh
rogeslagens.
61
Indogenous rogeslerone and exogenous rogeslagens in lhese
bilches resuIl in excessive GH secrelion
62
originaling from foci of hyerIaslic
ducluIar eilheIium of lhe mammary gIand.
63,64
The Ioss of GH uIsaliIily in bolh
humans and bilches has been ascribed lo lhe negalive feedback effecls of non-
eisodicaIIy secreled exlra-iluilary GH. Hovever, il is very unIikeIy lhal lhis
mechanism vas oeralionaI in lhe resenl dogs as onIy ovarieclomized bilches
vere used.

IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

95

The endocrine effecls of GH can be divided inlo raid calaboIic aclions
mainIy due lo insuIin anlagonism and in sIov anaboIic aclions. The sIov
hyerlrohic effecls of GH are medialed via IGI-I. In lhis resecl il is inleresling lo
nole lhal chiIdren vilh rimary hyolhyroidism have Iov Iasma IGI-I IeveIs
comared vilh concenlralions during lheray vilh L-lhyroxine.
34
Reduced
circuIaling IGI-I IeveIs have aIso been reorled in hyolhyroid rals.
20,65
The Iov
circuIaling IGI-I IeveIs robabIy refIecl lhe reduced GH secrelion in lhese chiIdren
and rals. In conlrasl, lhe resuIls of lhis sludy demonslrale lhal lhe Iasma IGI-I
IeveIs in dogs vilh rimary hyolhyroidism vere significanlIy higher lhan lhose
in lhe eulhyroid slale. The significanlIy eIevaled Iasma IGI-I IeveIs indicale lhal
lhe eIevaled GH secrelion in lhe hyolhyroid dogs has syslemic endocrine effecls.
Anolher indicalion lhal lhe eIevaled GH secrelion has endocrine significance
concerns lhe hysicaI changes observed in dogs vilh rimary hyolhyroidism.
olh in dogs vilh acromegaIy
41
and in dogs vilh rimary hyolhyroidism, lhe
hysicaI fealures are characlerized by lhick skin foIds. In acromegaIy lhe skin
lhickening has been ascribed lo mucooIysaccharide accumuIalion.
66
SimiIarIy, in
dogs vilh rimary hyolhyroidism lhe lhickening of lhe skin is aIso due lo
mucooIysaccharide accumuIalion.
67,68
Thus, some of lhe hysicaI changes
observed in dogs vilh rimary hyolhyroidism may very veII be acromegaIic
changes.
To summarize, lhe resuIls of lhis sludy demonslrale lhal rimary
hyolhyroidism in dogs is associaled vilh eIevaled basaI GH secrelion and Iess
GH secreled in uIses. This eIevaled GH secrelion has endocrine significance as
iIIuslraled by eIevaled Iasma IGI-I IeveIs and some hysicaI changes mimicking
acromegaIy. Il is discussed lhal lhe increased GH reIease in hyolhyroid dogs is
lhe resuIl of lhe absence of a resonse eIemenl for lhyroid hormone al lhe canine
iluilary GH gene and aIleralions in sura-iluilary reguIalion.

Acknnw!cdgmcnts

We are gralefuI for lhe lechnicaI assislance of Mrs. M. I. van WoIferen, Mrs.
C. I. M. Timmermans-Srang, Mrs. A. van Wees, Mrs. Y. M. I. A. IoIIak, and Mr. H.
G. H. van IngeIen.






Chaler 5
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"*

1. Harlman ML, CIaylon II, }ohnson ML, CeIniker A, IerIman A}, AIberli KG and Thorner MO
1993 A Iov dose eugIycemic infusion of recombinanl human insuIin-Iike grovlh faclor I
raidIy suresses fasling-enhanced uIsaliIe grovlh hormone secrelion in humans. } CIin
Invesl 91:2453-2462.
2. ermann M, }affe CA, Tsai W, DeMoll-Iriberg R and arkan AL 1994 Negalive feedback
reguIalion of uIsaliIe grovlh hormone secrelion by insuIin-Iike grovlh faclor I: InvoIvemenl
of hyolhaIamic somaloslalin. } CIin Invesl 94:138-145.
3. Ko|ima M, Hosoda H, Dale Y, Nakazalo M, Malsuo H and Kangava K 1999 GhreIin is a
grovlh-hormone-reIeasing acyIaled elide from slomach. Nalure 402:656-660.
4. Arval I, Camanni I and Ghigo I 1997 Age-reIaled grovlh hormone-reIeasing aclivily of
grovlh hormone secrelagogues in humans. Acla Iaedialr SuI 423:92-96.
5. Coras I, Harman SM and Iackman MR 1993 Human grovlh hormone and human aging.
Indocr Rev 14:20-39.
6. Van den erg G, VeIdhuis }D, IroIich M and RoeIfsema I 1996 An amIilude-secific
divergence in lhe uIsaliIe mode of grovlh hormone (GH) secrelion underIies lhe gender
difference in mean GH concenlralions in men and remenoausaI vomen. } CIin IndocrinoI
Melab 81:2460-2467.
7. Iaria AC, ekenslein LW, oolh RA }r, Vaccaro VA, AsIin CM, VeIdhuis }D, Thorner MO &
Ivans WS 1992 IuIsaliIe grovlh hormone reIease in normaI vomen during lhe menslruaI
cycIe. CIin IndocrinoI 36:591-596.
8. MendIevicz }, Linkovski I, Kerkhofs M, LerouIl R, Coinschi G and Van Cauler I 1999
Genelic conlroI of 24-hour grovlh hormone secrelion in man: a lvin sludy. } CIin IndocrinoI
Melab 84:856-862.
9. RiedeI M, Hoefl , Ium WI, von zur MuhIen A and rabanl G 1995 IuIsaliIe grovlh
hormone secrelion in normaI-veighl and obese men: differenliaI melaboIic reguIalion during
energy reslriclion. MelaboIism 44:605-610.
10. Malsuno M, Sohmiya M, Yamamolo H and Kalo Y 1998 An acromegaIic alienl vilh uIsaliIe
secrelion of grovlh hormone (GH) coincidenl vilh lhe sIov-vave sIee. Indocr } 45:687-692.
11. Irohman LA, Dovns TR and Chomczynski I 1992 ReguIalion of grovlh hormone secrelion.
Ironl NeuroendocrinoI 13:344-405.
12. Ahmad I, SleggIes AW, CarriIIo A} and IinkeIslein }A 1989 Obesily- and sexreIaled aIleralions
in grovlh hormone messenger RNA IeveIs. MoI CeII IndocrinoI 65:103-109.
13. Devesa }, Lima L and Tresguerres }AI 1992 Neuroendocrine conlroI of grovlh hormone
secrelion in humans. Trends IndocrinoI Melab 3:175-183.
14. Marlin D, IeIbaum }, Iuel-Ia|ol MT, IreIol M, Kordon C and Durand D 1985
Thyroideclomy aboIishes uIsaliIe grovlh hormone secrelion vilhoul affecling hyolhaIamic
somaloslalin. NeuroendocrinoIogy 41:476-481.

IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

97

15. ruhn TO, McIarIane M, Deckey }I and }ackson IM 1992 AnaIysis of uIsaliIe secrelion of
lhyrolroin and grovlh hormone in lhe hyolhyroid ral. IndocrinoIogy 131:2615-2621.
16. VareIa C, Cacicedo L, Iernandez G, de Ios IraiIes T and Sanchez Iranco I 1991 InfIuence of
hyolhyroidism duralion on deveIomenlaI changes in lhe hyolhaIamic faclors imIicaled
in grovlh hormone secrelion in lhe maIe ral. NeuroendocrinoIogy 54:340-345.
17. Sanlini I, Hurd RI, Lee and Chora I} 1993 Thyromimelic effecls of 3,5,3'-lriiodolhyronine
suIfale in hyolhyroid rals. IndocrinoIogy 133:105-110.
18. Tam SI, Lam KS and Srivaslava G 1996 Gene exression of hyolhaIamic somaloslalin,
grovlh hormone reIeasing faclor, and lheir iluilary recelors in hyolhyroidism.
IndocrinoIogy 137:418-424.
19. Mizobuchi M, Ishikava M, Okauchi Y, ando H and Sailo S 1996 Iffecls of lhyroideclomy on
lhyrolroin-reIeasing hormone (TRH) and somalolroin reIease-inhibiling faclor (SRIH)
allerns in inlrahyohysiaI microdiaIysales in rals. Indocr } 43:679-687.
20. Osorio A, Ruiz I & Orlega I 1998 IossibIe roIe of GH/IGI-1 in lhe ovarian funclion of aduIl
hyolhyroid rals. MoI CeII iochem 179:7-11.
21. Takeuchi A, Suzuki M and Tsuchiya S 1978 Iffecl of lhyroideclomy on lhe secrelory rofiIes
of grovlh hormone, lhyrolroin and corlicoslerone in lhe ral. IndocrinoIogy 25:381-390.
22. Kalakami H, Dovns TR and Irohman LA 1986 Decreased hyolhaIamic grovlh hormone-
reIeasing hormone conlenl and iluilary resonsiveness in hyolhyroidism. } CIin Invesl
77:1704-1711.
23. WiIIiams T, Maxon H, Thorner MO and Irohman LA 1985 Iunled grovlh hormone (GH)
resonse lo GH-reIeasing hormone inhyolhyroidism resoIves in lhe eulhyroid slale. } CIin
IndocrinoI Melab 61:454-456.
24. MarlinoIi MG and IeIIelier G 1989 Thyroid and gIucocorlicoid hormone reguIalion of ral
iluilary grovlh hormone messenger ribonucIeic acid as reveaIed by in silu hybridizalion.
IndocrinoIogy 125:1246-1252.
25. LIoyd RV, }in L, Song }Y, Terry LC, Horvalh I and Kovacs K 1990 Iffecls of royIlhiouraciI
on grovlh hormone and roIaclin messenger ribonucIeic acids in lhe ral iluilary. Lab Invesl
62:347-354.
26. Szabo M, Rueslov IC and Kramer DI 1985 Grovlh hormone resonse lo lhyrolroin-
reIeasing hormone in lhe urelhane-aneslhelized ral: effecl of lhyroid slalus. IndocrinoIogy
117:330-337.
27. Wood DI, IrankIyn }A, Docherly K, Ramsden D and Sheard MC 1987 The effecl of
lhyroid hormones on grovlh hormone gene exression in vivo in rals. } IndocrinoI 112:459-
463.
28. Korylko AI and CullIer L 1997 Thyroid hormone and gIucocorlicoid reguIalion of iluilary
grovlh hormone-reIeasing hormone recelor gene exression. } IndocrinoI 152:13-17.
29. Rool }L, Duckell GI, SveelIand M, SlrzeIecki }A and Rool AW 1985 Hyolhyroidism bIunls
lhe grovlh hormone (GH) reIeasing effecl of human ancrealic GH reIeasing faclor in lhe
aduIl maIe ral and . IndocrinoIogy 116:1703-1706.
Chaler 5
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98

30. Dam|anovic S, Ioovic V, Ielakov M, D|urovic M, Dieguez C & Casanueva II 1996 Iiluilary
enIargemenl due lo rimary hyolhyroidism: grovlh hormone resonse lo GHRH, GHRI-6
and GHRH Ius GHRI-6. } Iedialr IndocrinoI Melab 9:549-553.
31. IimenleI-IiIho IR, Ramos-Dias }C, Ninno I, Iacanha CI, Liberman and LengyeI AM 1997
Grovlh hormone resonses lo GH-reIeasing elide (GHRI-6) in hyolhyroidism. CIin
IndocrinoI 46:295-300.
32. uchanan CR, Slanhoe R, AdIard I, }ones }, Granl D and Ireece MA 1988 Gonadolrohin,
grovlh hormone and roIaclin secrelion in chiIdren vilh rimary hyolhyroidism. CIin
IndocrinoI 29:427-436.
33. Nishi Y, Hamamolo K, Ka|iyama M, Iu|ila A, Kavamura I, Kagava Y, Ka|ima T, Yamanaka M
and Uozumi T 1989 Iiluilary enIargemenl, hyerlrichosis and bIunled grovlh hormone
secrelion in rimary hyolhyroidism. Acla Iaedialr Scand 78:136-140.
34. Chernausek SD and Turner R 1989 Allenualion of sonlaneous, noclurnaI grovlh hormone
secrelion in chiIdren vilh hyolhyroidism and ils correIalion vilh Iasma insuIin-Iike grovlh
faclor I concenlralions. } Iedialr 114:968-972.
35. Chomczynski I, Soszynski IA and Irohman LA 1993 SlimuIalory effecl of lhyroid hormone
on grovlh hormone gene exression in a human iluilary ceII Iine. } CIin IndocrinoI Melab
77:281-285.
36. Lamberls SW, Ooslerom R, VerIeun T, Krenning II and Assies H 1984 ReguIalion of hormone
reIease by cuIlured ceIIs from a lhyrolroin-grovlh hormone-secreling iluilary lumor.
Direcl inhibiling effecls of 3,5,3'-lriiodolhyronine and dexamelhasone on lhyrolroin
secrelion. } IndocrinoI Invesl 7:313-317.
37. Moroval A and Dauncey M} 1998 Iffecls of lhyroid slalus on insuIin-Iike grovlh faclor-I,
grovlh hormone and insuIin are modified by food inlake. Iur } IndocrinoI 138:95-103.
38. IIsasser TH, Rumsey TS and Norlon SA 1992 ReIalionshis belveen lhe lhyroid and
somalolroic axes in sleers I: Iffecls of royIlhiouraciI-induced hyolhyroidism on grovlh
hormone, lhyroid slimuIaling hormone and insuIin-Iike grovlh faclor I. Domesl AnimaI
IndocrinoI 9:261-271.
39. IIelcher TI and CIarke I} 1994 Iffecls of hyolhyroidism on lhe grovlh hormone axis in
shee. } IndocrinoI 140:495-502.
40. IeIdman IC and NeIson RW 1996 Hyolhyroidism. In: Canine and IeIine IndocrinoIogy and
Reroduclion. 2
nd
edilion. IeIdman IC and NeIdon RW (Ids.) W. . Saunders Co.,
IhiIadeIhia. 68-117.
41. Ri|nberk A 1996 HyolhaIamus-iluilary syslem. In: CIinicaI IndocrinoIogy of Dogs and Cals.
Ri|nberk A (Id.). Dordrechl/oslon: KIuver Academic IubIishers. I:11-34.
42. runer }M, Scoll-Moncrieff }C and WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am
Vel Med Assoc 15:1572-1575.
43. Iigenmann }I and Iigenmann RY 1981 Radioimmunoassay of canine grovlh hormone. Acla
IndocrinoI 98:514-520.
IH in dogs is associaled vilh eIevaled GH reIease
_________________________________________________________________________________

99

44. Na RC, MoI }A and HazevinkeI HAW 1993 Age-reIaled Iasma concenlralions of grovlh
hormone (GH) and insuIin-Iike grovlh faclor 1(IGI-1) in Greal Dane us fed differenl
dielary IeveIs of rolein. Domes AnimaI IndocrinoI 10:237-247.
45. Merriam GR and Wachler KW 1982 AIgorilhms for lhe sludy of eisodic hormone secrelion.
Am } IhysioI 243:310-318.
46. Love|oy }C, Smilh SR, ray GA, VeIdhuis }D, Rood }C and TuIIey R 1997 Iffecls of
exerimenlaIIy induced miId hyerlhyroidism on grovlh hormone and insuIin secrelion and
sex sleroid IeveIs in heaIlhy young men. MelaboIism 46:1424-1428.
47. HoIIenberg AN, Monden T, IIynn TR, oers MI, Cohen O & Wondisford II 1995 The human
lhyrolroin-reIeasing hormone gene is reguIaled by lhyroid hormone lhrough lvo dislincls
cIasses of negalive lhyroid hormone resonse eIemenls. MoI IndocrinoI 9:540-550.
48. reen }}, Hickok N} & Gurr }A 1995 The ral TSHbela gene conlains dislincl resonse eIemenls
for reguIalionby relinoids and lhyroid hormone. MoI CeII IndocrinoI 131:137-146.
49. Rado|a N, Diaz DV, Minars T}, Ireedberg IM, Iumenberg M & Tomic-Canic M 997 Secific
organizalion of lhe negalive resonse eIemenls for relinoic acid and lhyroid hormone
recelors in keralin gene famiIy. } Invesl Dermal 109:566-572.
50. Lanlinga-van Leeuven IS and MoI }A 2000 CIoning and characlerizalion of lhe 5' fIanking
region of lhe canine grovlh hormone gene. In: ReguIalion of grovlh hormone gene
exression in lhe canine mammary gIand. Thesis of I.S. Langlinga-van Leeuven, Ulrechl
Universily. :81-90.
51. Iaggiano M, CriscuoIo T, Graziani M, Iorio S, Iisano G, Sinisi AA, Sinisi AM and eIIasleIIa A
1985 Iersislenl TRH-induced grovlh hormone reIease afler shorl-lerm and Iong-lerm L-
lhyroxine reIacemenl lheray in rimary congenilaI hyolhyroidism. CIin IndocrinoI 23:61-
66.
52. Coiro V, VoIi R, Carelli L, Seroni, Marchesi C, Vescovi G, Caffarri G, CoIIa R, Rossi G,
DavoIi C and Chiodera I 1994 InfIuence of lhyroid slalus on lhe aradoxicaI grovlh hormone
resonse lo lhyrolroin-reIeasing hormone in human obesily. MelaboIism 43:514-517.
53. Hanev K, Ulsumi A, Sugavara A, Tanaka A, Iukazava H, Yoshida K and Abe K. 1995
Inhanced Iasma GH resonses lo simuIlaneous adminislralion of TRH and GHRH in
alienls vilh rimary hyolhyroidism. Indocr } 42:43-47.
54. Konaka S, Yamada M, Saloh T, Ozava H, Walanabe I, Takala and Mori M 1997 Ixression of
lhyrolrohin-reIeasing hormone (TRH) recelor mRNA in somalolrohs in lhe ral anlerior
iluilary. IndocrinoIogy 138:827-830.
55. aIdini M, Calania A, Orsalli A, Manfredi MG, Molla I and CanlaIamessa L 1992 Inhibilory
effecl of somaloslalin on abnormaI GH resonse lo TRH in rimary hyolhyroidism. Ix
CIin IndocrinoI 99:80-83.
56. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IM and
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114:275-282.
Chaler 5
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57. Takahashi Y, Ibihara S, Nakamura Y and Takahashi K 1981 A modeI of human sIee-reIaled
grovlh hormone secrelion in dogs: effecls of 3, 6, and 12 hours of forced vakefuIness on
Iasma grovlh hormone, corlisoI, and sIee slages. IndocrinoIogy 109:262-272.
58. Irench M, Vailkus I, Cukerman I, Sirek A and Sirek OV 1987 Secrelory allern of canine
grovlh hormone. Am } IhysioI 252:268-272.
59. Kooislra HS, den Herlog I, Okkens AC, MoI }A and Ri|nberk A 2000 IuIsaliIe secrelion
allern of grovlh hormone during lhe IuleaI hase and mid-anoeslrus in beagIe bilches. }
Rerod IerliI 119:217-222.
60. Iriksson L, Irankenne I, Iden S, Hennen G and Von SchouIlz 1989 Grovlh hormone 24-h
serum rofiIes during regnancy-Iack of uIsaliIily for lhe secrelion of lhe IacenlaI varianl.
r } Obslel GynaecoI 96:949-953.
61. Walson AD}, Rulleman GR, Ri|nberk A & MoI }A 1987 Iffecl of somaloslalin anaIogue SMS
201-995 and anlirogeslin agenl RU486 in canine acromegaIy. Ironl Horm Res 17:193-198.
62. Iigenmann }I, Iigenmann RY, Ri|nberk A, van der Gaag I, Zaf } and Iroesch IR 1983
Irogeslerone-conlroIIed grovlh hormone overroduclion and naluraIIy occurring canine
diabeles and acromegaIy. Acla IndocrinoI 104:167-176.
63. SeIman I}, MoI }A, Rulleman GR, van Garderen I & Ri|nberk A 1994 Irogeslin-induced
grovlh hormone excess in lhe dog originales in lhe mammary gIand. IndocrinoIogy 134:287-
292.
64. Van Garderen I, de Wil M, Voorhoul WI, Rulleman GR, MoI }A, Nederbragl H and Misdor
W 1997 Ixression of grovlh hormone in canine mammary lissue and mammary lumors.
Ividence for a olenliaI aulocrine/aracrine slimuIalory Ioo. Am } IalhoI 150:1037-1047.
65. Ramos S, Goya L, AIvarez C and IascuaI-Leone AM 1998 Mechanism of hyolhyroidism
aclion on insuIin-Iike grovlh faclor-I and -II from neonalaI lo aduIl rals: insuIin mediales
lhyroid hormone effecls in lhe neonalaI eriod. IndocrinoIogy 139:4782-4792.
66. Malsuoka LY, Worlsman }, KucheIIa CI, Ing A and Dielrich }I 1982 HislochemicaI
characlerizalion of lhe culaneous invoIvemenl of acromegaIy. Arch Inlern Med 142:1820-1823.
67. MuIIer GH, Kirk RW and Scoll DW 1989 Culaneous endocrinoIogy. In: SmaII AnimaI
DermaloIogy. 4
lh
edilion. MuIIer GH, Kirk RW and Scoll DW (Ids). IhiIadeIhia:W
Saunders Co. : 575-657.
68. DoIiger S, DeIverdier M, More }, Longearl L, Regnier A and MagnoI }I 1995 HislochemicaI
sludy of culaneous mucins in hyolhyroid dogs. Vel IalhoI 32:628-34.

Chaptcr 6

Thyrntrnpin-rc!casing hnrmnnc-induccd
grnwth hnrmnnc sccrctinn in
dngs with primary hypnthyrnidism








M.M. Diaz-Espincira, 5. Ga!ac, J.A. Mn!, A. Ri|nbcrk, H.5. Knnistra



Dnmcstic Anima! Endncrinn!ngy 2008, 34: 176-181



Dearlmenl of CIinicaI Sciences of Comanion AnimaIs
IacuIly of Velerinary Medicine, Ulrechl Universily, Ulrechl, The NelherIands.


Chaler 6
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102

Abstract

Irimary hyolhyroidism in dogs is associaled vilh increased reIease of
grovlh hormone (GH). In search for an exIanalion ve invesligaled lhe effecl of
inlravenous adminislralion of lhyrolroin-reIeasing hormone (TRH, 10 g/kg body
veighl) on GH reIease in 10 dogs vilh rimary hyolhyroidism and 6 heaIlhy
conlroI dogs. The hyolhyroid dogs had a medicaI hislory and hysicaI changes
comalibIe vilh hyolhyroidism and vere incIuded in lhe sludy on lhe basis of
lhe foIIoving crileria: Iasma lhyroxine concenlralion < 2 nmoI/I and Iasma
lhyrolroin (TSH) concenlralion > 1 g/I. In addilion,
99m
TcO4
-
ulake during
lhyroid scinligrahy vas Iov or absenl.
TRH adminislralion caused Iasma TSH concenlralions lo rise
significanlIy in lhe conlroI dogs, bul nol in lhe hyolhyroid dogs. In lhe dogs vilh
rimary hyolhyroidism, lhe mean basaI Iasma GH concenlralion vas reIaliveIy
high (2.3 0.5 g/I) and increased significanlIy (I0.001) 10 and 20 min afler
in|eclion of TRH (lo 11.9 3.5 g/I and 9.8 2.7 g/I, resecliveIy). In lhe conlroI
dogs, lhe mean basaI Iasma GH concenlralion vas 1.3 0.1 g/I and did nol
increase significanlIy afler TRH adminislralion.
We concIude lhal, in conlrasl lo heaIlhy conlroI dogs, rimary
hyolhyroid dogs resond lo TRH adminislralion vilh a significanl increase in lhe
Iasma GH concenlralion, ossibIy as a resuIl of lransdifferenlialion of
somalolroic iluilary ceIIs lo lhyrosomalolroes.


TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
103

Intrnductinn

Insufficienl IeveIs of lhyroid hormones affecl aImosl aII organ syslems,
incIuding olher endocrine gIands such as lhe iluilary gIand.
1
The effecl of
hyolhyroidism on lhe reIease of iluilary hormones varies er secies and lhis is
arlicuIarIy lrue for grovlh hormone (GH). In some mammaIs, such as shee and
sleers, (exerimenlaIIy induced) rimary hyolhyroidism does nol affecl GH
secrelion,
2,3
vhereas in rals and igs hyolhyroidism Ieads lo Iov Iasma GH
concenlralions.
4-9
In conlrasl, in dogs rimary hyolhyroidism is associaled vilh
eIevaled basaI GH secrelion.
10
AIlhough lhe uIsaliIe characler of GH reIease is
mainlained, Iess GH is reIeased er uIse. Hovever, inlegraI GH reIease caIcuIaled
as area under lhe curve for GH above zero IeveI is significanlIy eIevaled in
hyolhyroid dogs. This eIevaled GH secrelion has endocrine effecls as refIecled by
eIevaled Iasma IGI-I concenlralions and hysicaI changes mimicking
acromegaIy.
10

In lhe search for an exIanalion for lhis unique silualion in dogs, il has
been suggesled lhal lhe Iov Iasma lhyroid hormone concenlralion affecls lhe
reguIalion of GH reIease al lhe hyolhaIamic IeveI. In heaIlhy humans
adminislralion of lhyrolroin-reIeasing hormone (TRH) does nol resuIl in
areciabIe changes in Iasma GH IeveIs,
11-13
vhereas in humans vilh rimary
hyolhyroidism TRH adminislralion resuIls in a significanl increase in lhe Iasma
GH concenlralion.
4,11-13
This aradoxicaI resonse of GH lo TRH in rimary
hyolhyroidism may in arl be caused by lhe reIalive deficiency in
somaloslalin.
6,7,10
Infusion of somaloslalin inhibils lhe GH resonse lo TRH in
humans vilh rimary hyolhyroidism.
14

So far, lhere is no informalion on lhe effecl of TRH on GH secrelion in
dogs vilh rimary hyolhyroidism. Here ve reorl on lhe effecl of TRH
adminislralion on lhe Iasma GH concenlralion in heaIlhy conlroI dogs and in
dogs vilh rimary hyolhyroidism.

Matcria!s and Mcthnds


Ten cIinicaI cases, resenled vilh a medicaI hislory and hysicaI changes
comalibIe vilh hyolhyroidism, vere incIuded in lhe sludy on lhe basis of lhe
foIIoving crileria: Iasma lhyroxine (T4) concenlralion < 2 nmoI/I and Iasma
lhyrolroin (TSH) concenlralion > 1 g/I (TabIe 1). In addilion,
99m
TcO4
-
ulake
during lhyroid scinligrahy vas Iov or absenl.
15
There vere six maIe dogs (4
caslraled) and four femaIe dogs (2 sayed) of a vide variely of breeds, lhe median
age vas 6 years (range 3 11 years) (TabIe 1). According lo lhe ovners, lhe
Chaler 6
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104

hysicaI changes comalibIe vilh hyolhyroidism had been resenl for severaI
monlhs (range 6 lo 18 monlhs).
In addilion, six heaIlhy beagIe bilches (median age 5 years, range 27
years) vere sludied during anoeslrus as conlroI dogs. Their Iasma concenlralions
of T4 and TSH vere vilhin lhe reference ranges of lhese hormones. The dogs vere
housed in airs in indoor-ouldoor runs, fed on a slandard manufaclured dog food
lvice daiIy, and given valer a! |i|iiun

Tcsi an! sanp|c cc||cciicn
Iood samIes for GH measuremenls vere coIIecled al 15, 0, 5, 10, 20, 30,
45 min afler inlravenous adminislralion of 10 g/kg of TRH (TRH UC,
IrolireIinum, S.A. UC N.V., Secleur Iharma, raine-IAIIeud, eIgium), and
samIes for TSH measuremenl vere coIIecled al 15, 0, and 30 min. The samIes
vere coIIecled by |uguIar veniunclure, immedialeIy lransferred inlo chiIIed
IDTA-coaled (GH) or hearin-coaled (TSH) lubes, and cenlrifuged in a cooIed
cenlrifuge. IIasma vas slored al 20C unliI assayed.

Hcrncnc ncasurcncnis
IIasma TSH concenlralions vere delermined vilh a homoIogous soIid-
hase, lvo-sile chemiIuminescence enzyme immunomelric assay

(ImmuIile canine
TSH, Diagnoslic Iroducls Cororalion, Los AngeIes, CA, U.S.A) in accordance
vilh lhe inslruclions of lhe manufaclurer. The inlra-assay coefficienls of varialion
vere 5.0, 4.0 and 3.8% al TSH IeveIs of 0.20, 0.50 and 2.6 g/I, resecliveIy. The
inler-assay coefficienls of varialion vere 6.3 and 8.2% al TSH IeveIs of 0.16 and 2.8
g/I, resecliveIy. The Iovesl deleclabIe amounl of TSH vas 0.03 g/I.
16

IIasma T4 concenlralions vere delermined vilh a homoIogous soIid-
hase, chemiIuminescence enzyme immunoassay (ImmuIile TolaI T4, Diagnoslic
Iroducls Cororalion, Los AngeIes, CA, U.S.A.) in accordance vilh lhe
inslruclions of lhe manufaclurer. The inlra-assay coefficienls of varialion vere 13.8
and 8.2% al T4 IeveIs of 8 and 25 nmoI/I, resecliveIy.
16
The inler-assay coefficienl
of varialion vas 8.5% al T4 IeveIs of 21.0 nmoI/I. The Iovesl deleclabIe amounl of
T4 vas 2 nmoI/I.
IIasma GH concenlralions vere delermined vilh a commerciaIIy avaiIabIe
radioimmunoassay for orcine and canine GH (IGH-46HK, Linco Research, Sl.
CharIes, MS, U.S.A). The degree of cross-reaclion vilh roIaclin in lhe canine GH
assay vas < 0.1% . The inlra-assay coefficienl of varialion vas 7.6% al a Iasma
concenlralion of 4.4 g/I. The inler-assay coefficienl of varialion vas 10% al Iasma
concenlralion of 4.3 g/I. The sensilivily of lhe assay vas 1 g/I.


TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
105

Dng Brccd Gcndcr Agc
(ycars)
Bndy
wcight
(kg)
T4
(nmn!/!)
T5H
(g/!)
1 Cross-
bred
CM 9 22.4 < 2 2.50
2 }ack
RusseI
CI 6 10.0 < 2 2.50
3 Hovavarl

I 4 32.5 < 2 2.40
4 ShelIand
Sheedog
M 5 12.9 < 2 2.50
5 Cross-
bred
CM 5 31.4 < 2 1.60
6 WeIsh
Sringer
SanieI
M 6 25.5 < 2 4.3
7 Labrador
Relriever
CM 9 38 < 2 3.10
8 ShelIand
Sheedog
CM 11 19.6 < 2 1.80
9 GoIden
Relriever
CI 5 40.9 < 2 9.50
10 Cairn
Terrier
I 3 10.6 < 2 5.50
RcIcrcncc va!ucs 17-46 < 0.6




Tab!c 1. SignaImenls and resenling Iasma T4 and TSH vaIues in 10 dogs vilh
rimary hyolhyroidism. M: maIe, I: femaIe, CM: caslraled maIe, CI: caslraled
femaIe.



Chaler 6
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106

Oaia ana|qsis
The normaIily of lhe dala vas assessed vilh lhe KoImogorov-Smirnov lesl.
The unaired Sludenl's i-lesl vas used lo comare basaI IeveIs of GH and TSH
belveen lhe hyolhyroid grou and lhe conlroI grou. The aired Sludenl's i-lesl
vas used lo comare basaI Iasma TSH concenlralions vilh lhe Iasma TSH
concenlralions 30 min afler TRH adminislralion. The ANOVA for reealed
measuremenls (osl-hoc lesl: Tukey lesl) vas used lo invesligale if lhere vas a
significanl increase in GH afler TRH adminislralion. Ior aII comarisons I 0.05
vas considered significanl.

Ethics

The design of lhis sludy vas aroved by lhe Ilhics Commillee of lhe
IacuIly of Velerinary Medicine, Ulrechl Universily.

Rcsu!ts

In lhe TRH-slimuIalion lesl lhe basaI Iasma TSH concenlralions of lhe
heaIlhy dogs vere vilhin lhe reference range (< 0.6 g/I). Adminislralion of TRH
caused lhe Iasma TSH concenlralions lo rise significanlIy (Iig 1) from 0.35 0.05
g/I lo 1.8 0.4 g/I (I0.025). SimiIar lo lhe iniliaI vaIues on admission, in lhe
hyolhyroid dogs lhe basaI Iasma TSH concenlralions of lhe TRH-slimuIalion lesl
(3.1 + 0.6 g/I) exceeded lhe uer Iimil of lhe reference range and vere
significanlIy higher (I0.005) lhan lhose of lhe conlroIs. Adminislralion of TRH
had no slalislicaIIy significanl effecl (I0.08) on lhe Iasma TSH concenlralions of
lhe hyolhyroid dogs (Iig 1). Thirly minules afler TRH adminislralion lhe Iasma
TSH concenlralion vas nol significanlIy differenl in lhe hyolhyroid and conlroI
dogs (I0.12).
The mean basaI Iasma GH concenlralion vas higher in lhe dogs vilh
rimary hyolhyroidism lhan in lhe conlroI dogs (2.3 0.5 g/I versus 1.3 0.1
g/I), bul lhe difference did nol reach significance (I0.07). In lhe hyolhyroid
dogs lhe mean Iasma GH concenlralions had increased significanlIy (I0.001) 10
and 20 min afler TRH adminislralion (11.9 3.5 g/I and 9.8 2.7 g/I,
resecliveIy), bul lhis vas nol lhe case in lhe conlroI dogs (I0.29) (Iig 2).






TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
107







Figurc 1. IIasma TSH resonse (mean SIM) of six heaIlhy beagIe dogs ( --) and
10 dogs vilh rimary hyolhyroidism (--). Al lime oinl 0 TRH vas adminislered
inlravenousIy in a dosage of 10 g er kg body veighl. significanlIy higher lhan
baseIine vaIues.

Chaler 6
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108



































Figurc 2. IIasma GH concenlralion (mean SIM) of six heaIlhy beagIe dogs (--)
and 10 dogs vilh rimary hyolhyroidism ( -- ). Al lime oinl 0 TRH vas
adminislered inlravenousIy in a dosage of 10 g er kg body veighl.
significanlIy higher lhan baseIine vaIues.

TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
109



Il can be difficuIl lo eslabIish lhe diagnosis of rimary hyolhyroidism in
dogs because of lhe common occurrence of Iov Iasma T4 concenlralions in non-
lhyroidaI iIIness and lhe frequenl absence of an eIevaled Iasma TSH concenlralion
in rimary hyolhyroidism.
17
In order lo revenl incIusion of misdiagnosed cases,
ve enlered onIy dogs in vhich lhe resence of rimary hyolhyroidism vas
beyond any doubl. AII dogs had a reIaliveIy Iong hislory of hysicaI changes
comalibIe vilh hyolhyroidism. The Iasma T4 concenlralions vere exlremeIy
Iov and lhe Iasma TSH concenlralions vere raised. In addilion, lhyroidaI
99m
TcO4
-

ulake vas Iov or absenl. This seIeclion for unequivocaI cases may have
infIuenced lhe resuIls of lhe TRH-slimuIalion lesls.
In lhe heaIlhy dogs lhe TSH increase (median 331%, range 1701064%)
afler TRH slimuIalion vas comarabIe lo lhal reorled reviousIy (median 207%,
range 252200%).
18
In lhe hyolhyroid dogs lhe median osl-slimuIalion TSH
concenlralion vas onIy 6 % (range 821%) higher lhan lhe averaged basaI vaIues.
In a revious sludy a median rise of 24% (range 21134%) vas found.
18
The
hyolhyroid slale affecls lhe resonsiveness of TSH lo slimuIalion vilh TRH, and
lhis may have been even more so in lhis grou of ronounced cases.
Wilh regard lo GH ve observed lhe reverse. In lhe heaIlhy dogs Iasma
GH concenlralions did nol increase significanlIy afler TRH adminislralion, vhich
is in agreemenl vilh lhe resuIls of an earIier sludy.
19
In conlrasl, in lhe dogs vilh
rimary hyolhyroidism lhere vas a slrong resonse of lhe Iasma GH
concenlralions lo TRH adminislralion. This observalion exlends earIier findings of
eIevaled basaI Iasma GH concenlralions and aIleralions in lhe uIsaliIe secrelion
allern of GH in dogs vilh rimary hyolhyroidism.
10

Thus rimary hyolhyroidism in dogs Ieads lo lvo abnormaI resonses lo
TRH adminislralion, nameIy a hyoresonse of TSH and a aradoxicaI resonse of
GH. Ior bolh hormones lhe basaI vaIues are oflen eIevaled. These abnormaIilies
viII be discussed aIong lhe Iines of lhe naluraI deveIomenl of lhe disease.
In agreemenl vilh lhe cIassic endocrine rinciIes of feedback reguIalion,
in rimary hyolhyroidism lhe decreasing lhyroid hormone secrelion increases lhe
reIease of TSH. The negalive feedback reguIalion of TSH secrelion by lhyroid
hormones is sensilive lo changes of 10% lo 20% in Iasma lhyroid hormone
concenlralion.
20
In human medicine an increased baseIine Iasma TSH
concenlralion confirms lhe diagnosis of rimary hyolhyroidism. Tesling vilh
TRH is nol arl of lhe diagnoslic vork-u in humans, aIlhough il is knovn lhal lhe
TSH resonse lo TRH is aIvays increased in humans vilh rimary
hyolhyroidism.
21
AIso alienls vilh Iong-slanding rimary hyolhyroidism and
hyerIasia of lhyrolroes resond in an exaggeraled fashion lo TRH
Chaler 6
_________________________________________________________________________________
110

slimuIalion.
22
In humans a bIunled TSH resonse lo slimuIalion vilh TRH seems
lo occur onIy in alienls vilh TSH-secreling iluilary lumors.
23
Thus lhe frequenl
occurrence of bIunled or absenl TSH resonses in lhe TRH lesl are secific lo
hyolhyroid dogs, and so far unexIained.
Wilh regard lo lhe aradoxicaI GH resonse afler TRH adminislralion,
sludies in humans and research in rals vilh exerimenlaIIy induced
hyolhyroidism indicale lhal lhe cIassic endocrine concel of one iluilary ceII-
one iluilary hormone shouId be abandoned. The discovery of bi-hormonaI ceIIs
has Ied lo lhe concel caIIed lransdifferenlialion, i.e., lhal on demand iluilary
ceIIs of one hormonaI lye may shifl lo anolher.
24
In rals made hyolhyroid by
royIlhiouraciI adminislralion, somalolroes lransform inlo slimuIaled
lhyrolroes or so-caIIed lhyroideclomy ceIIs. This lransdifferenlialion is
reversibIe since disconlinualion of royIlhiouraciI lrealmenl Ieads lo lhe
disaearance of lhyroideclomy ceIIs and lhe reaearance of somalolroes.
25
In
humans vilh rolracled rimary hyolhyroidism massive lhyrolroe hyerIasia
has been found. DoubIe immunoslaining, immunoeIeclron microscoy, and in silu
hybridizalion reveaIed lhe resence of bihormonaI ceIIs roducing bolh GH and
TSH.
24
Thus, in addilion lo lhe muIliIicalion of lhyrolroes, lhe
lransdifferenlialion of somalolroes inlo lhyrolroes conlribules lo lhe increase in
TSH-roducing ceIIs.
In rals vilh melhimazoIe-induced hyolhyroidism lhe number of TRH-
recelor osilive ceIIs increases in araIIeI vilh lhe number of TSH-
immunoreaclive ceIIs. This indicales TRH slimuIalion has a roIe in lhe
lransdifferenlialion rocess.
26
Il is aIso in Iine vilh lhe reversibIe characler of lhe
rocess uon cessalion of lhe slimuIalion, lhal is disconlinualion of lhe
adminislralion of lhe anlilhyroid drug in lhe exerimenlaI silualion or lhyroid
subslilulion lheray in case of sonlaneous rimary hyolhyroidism.
22

Againsl lhe background of lhis concel of lransdifferenlialion, lhal is lhe
henolyic svilch belveen differenl malure iluilary ceII lyes vilhoul ceII
division, il is lemling lo secuIale lhal a simiIar mechanism oerales in dogs vilh
rimary hyolhyroidism. The deveIomenl of lhe bihormonaI lhyrosomalolroic
ceIIs under lhe infIuence of ongoing TRH slimuIalion vouId exIain lhe eIevaled
basaI IeveIs of TSH and GH. AIso lhe aradoxicaI GH resonse lo TRH
adminislralion is consislenl vilh lhis concel. The oflen bIunled TSH resonse lo
TRH adminislralion seen in hyolhyroid dogs mighl be exIained by lhe
deveIomenl of lhyrosomalroic ceIIs vilh an aIlered osl-TRH-recelor
signaIIing alhvay for lhe synlhesis and/or reIease of TSH. Sludies are in rogress
lo lesl some of lhese assumlions.
Il can be concIuded lhal, in conlrasl lo heaIlhy dogs, rimary hyolhyroid
dogs resond lo TRH adminislralion vilh a significanl increase in lhe Iasma GH
TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
111

concenlralion, ossibIy as a resuIl of lransdifferenlialion of somalolroes lo
lhyrosomalolroes.

Acknnw!cdgmcnts

The lechnicaI assislence of Mrs. Y.W.I.A. IoIIak, Mr. H.G.H. van IngeIen,
Ms. }. WoIfsvinkeI, and Ms. M.I. van WoIferen is highIy arecialed.

Chaler 6
_________________________________________________________________________________
112

#:

1. Snyder I} 2000 The iluilary in hyolhyroidism. In: The Thyroid. A fundamenlaI and CIinicaI
Texl. 8
lh
edilion. raverman LI and Uliger RD (Ids.), Iighl Idilion, Liincoll WiIIians &
WiIkins. A WoIler KIuver Comany, IhiIadeIhia. :811-814.
2. IIsasser TH, Rumsey TS, Norlon SA 1992 ReIalionshis belveen lhe lhyroid and
somalolroic axes in sleers I: effecls of royIlhiouraciI-induced hyolhyroidism on grovlh
hormone, lhyroid slimuIaling hormone and insuIin-Iike grovlh faclor I. Domesl Anim
IndocrinoI 9:261-271.
3. IIelcher TI, CIarke I} 1994 Iffecls of hyolhyroidism on lhe grovlh hormone axis in shee. }
IndocrinoI 140:495-502.
4. VareIa C, Cacicedo L, Iernandez G, de Ios IraiIes T, Sanchez Iranco I 1991 InfIuence of
hyolhyroidism duralion on deveIomenlaI changes in lhe hyolhaIamic faclors imIicaled
in grovlh hormone secrelion in lhe maIe ral. NeuroendocrinoIogy 54:340-345.
5. Sanlini I, Hurd RI, Lee , Chora I} 1993 Thyromimelic effecls of 3,5,3'-lriiodolhyronine
suIfale in hyolhyroid rals. IndocrinoIogy 133:105-110.
6. Mizobuchi M, Ishikava M, Okauchi Y, ando H, Sailo S 1996 Iffecls of lhyroideclomy on
lhyrolroin-reIeasing hormone (TRH) and somalolroin reIease-inhibiling faclor (SRIH)
allerns in inlrahyohyseaI microdiaIysales in rals. Indocrine } 43:679-687.
7. Tam SI, Lam KS, Srivaslava G 1996 Gene exression of hyolhaIamic somaloslalin, grovlh
hormone reIeasing faclor, and lheir iluilary recelors in hyolhyroidism. IndocrinoIogy
137:418-424.
8. Osorio A, Ruiz I, Orlega I 1998 IossibIe roIe of GH/IGI-I in lhe ovarian funclion of aduIl
hyolhyroid rals. MoI CeII iochem 179:7-11.
9. Moroval A, Dauncey M} 1998 Iffecls of lhyroid slalus on insuIin-Iike grovlh faclor-I, grovlh
hormone and insuIin are modified by food inlake. Iur } IndocrinoI 138:95-103.
10. Lee WM, Diaz-Isineira MM, MoI }A, Ri|nberk A, Kooislra HS 2001 Irimary hyolhyroidism
in dogs is associaled vilh eIevaled GH reIease. } IndocrinoI 168:59-66.
11. Iaggiano M, CriscuoIo T, Graziani M, Iorio S, Iisano G, Sinisi AA, Sinisi AM, eIIasleIIa A
1985 Iersislenl TRH-induced grovlh hormone reIease afler shorl-lerm and Iong-lerm L-
lhyroxine reIacemenl lheray in rimary congenilaI hyolhyroidism. CIin IndocrinoI 23:61-
66.
12. Coiro V, VoIi R, Carelli L, Seroni, Marchesi C, Vescovi G, Caffari G, CoIIa R, Rossi G,
DavoIi C, Chiodera I 1994 InfIuence of lhyroid slalus on lhe aradoxicaI grovlh hormone
resonse lo lhyrolroin-reIeasing hormone in human obesily. MelaboIism 43:514-517.
13. Hanev K, Ulsumi A, Sugavara A, Tanaka A, Iukazava H, Yoshida K, Abe K 1995 Inhanced
Iasma GH resonses lo simuIlaneous adminislralion of TRH and GHRH in alienls vilh
rimary hyolhyroidism. Indocrine } 42:43-47.


TRH-induced GH secrelion in dogs vilh IH
_________________________________________________________________________________
113

14. aIdini M, Calania A, Orsalli A, Manfredi MG, Molla I, CanlaIamessa L 1992 Inhibilory effecl
of somaloslalin on abnormaI GH resonse lo TRH in rimary hyolhyroidism. Ix CIin
IndocrinoI 99: 80-83.
15. Diaz Isineira MM, MoI }A, Ieelers MI, Iversen LI, van Di|k }I, Ri|nberk A, Kooislra HS.
Assessmenl of lhyroid funclion in dogs vilh Iov Iasma lhyroxine concenlralion. } Vel Inl
Med (In Iress).
16. Kooislra HS, Diaz-Isineira M, MoI }A, van den rom WI, Ri|nberk A 2000 Secrelion allern
of lhyroid-slimuIaling hormone in dogs during eulhyroidism and hyolhyroidism. Domesl
Anim IndocrinoI 18:19-29.
17. Scoll-Moncrieff }CR 2005 Hyolhyroidism. In: Texlbook of Velerinary InlernaI Medicine. 6
lh

edilion. Illinger S} and IeIdman IC (Ids.). Sl. Louis, IIsevier Saunders. :1535-1544.
18. Scoll-Moncrieff }CR and NeIson RW 1998 Change in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolroin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and eulhyroid dogs vilh concurrenl disease. } Am Vel Med Assoc
213:1435-1438.
19. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IMM,
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114:275-282.
20. Ordene KW, Ian C, arzeI US, Surks MI 1983 VariabIe lhyrolroin resonse lo lhyrolroin-
reIeasing hormone afler smaII decreases in Iasma lhyroid hormone concenlralions in
alienls of advanced age. MelaboIism 32:881-888.
21. Shairo LI and Surks MI 2001 Hyolhyroidism. In : IrinciIes and Iraclice of IndocrinoIogy
and MelaboIism. 3rd edilion. ecker KL (Id.). IhiIadeIhia, Liincoll WiIIiams &
WiIkins.:445-454.
22. SarIis N}, rucker-Davis I, Doman }L, SkaruIis MC 1997 MRI-demonslrabIe regression of a
iluilary mass in a case of rimary hyolhyroidism afler a veek of acule lhyroid hormone
lheray. } CIin IndocrinoI Melab 82:808-811.
23. rucker-Davis I, OIdfieId IH, SkaruIis MC, Doman }L, Weinlrabu D 1999 Thyrolroin-
secreling iluilary lumors: Diagnoslic crileria, lhyroid hormone sensilivily, and lrealmenl
oulcome in 25 alienls foIIoved al lhe NalionaI Inslilules of HeaIlh. } CIin IndocrinoI Melab
84:476-486.
24. VidaI S, Horvalh I, Kovacs K, Cohen SM, LIoyd RV, Scheilhauer W 2000
Transdifferenlialion of somalolrohs lo lhyrolrohs in lhe iluilary of alienls vilh
rolracled rimary hyolhyroidism. Virchovs Arch 436:43-51.
25. Horvalh I, LIoyd RV, Kovacs K 1990 IroyIlhiouraciI-induced hyolhyroidism resuIls in
reversibIe lransdifferenalion of somalolrohs inlo lhyroideclomy ceIIs. A morhoIogic sludy
of lhe ral iluilary incIuding immunoeIeclron microscoy. Lab Invesl 63:511-520.


Chaler 6
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114

26. Radian S, CocuIescu M, Morris }I 2003 Somalolroh lo lhyrolroh ceII lransdifferenlialion
during exerimenlaI hyolhyroidism a Iighl and eIeclron-microscoy sludy. } CeII MeI Med
7:297-306.





Chaptcr 7

Functinna! and mnrphn!ngica! changcs
in thc adcnnhypnphysis nI dngs
with induccd primary hypnthyrnidism:
!nss nI T5H hypcrsccrctinn, hypcrsnmatntrnpism,
hypnprn!actincmia, and pituitary cn!argcmcnt
with transdiIIcrcntiatinn


M.M. Diaz Espincira
1
, J.A. Mn!
1
, T.5.G.A.M. van dcn Ingh
2
,
R.H. van dcr V!ugt-Mci|cr
3
, A. Ri|nbcrk
1
, H.5. Knnistra
1


Dnmcstic Anima! Endncrinn!ngy 2008, 35:98-111

_________________________________________________________________________

1
Dearlmenl of CIinicaI Sciences of Comanion AnimaIs

2
Dearlmenl of IalhobioIogy, and

3
Division of Diagnoslic Imaging
IacuIly of Velerinary Medicine, Ulrechl Universily
Ulrechl, The NelherIands
Chaler 7
_________________________________________________________________________________

116

%/

Irom case sludies in humans il is knovn lhal rimary hyolhyroidism
(IH) may be associaled vilh morhoIogicaI and funclionaI changes of lhe
iluilary. There is no insighl inlo lhe lime scaIe of lhese changes. In lhis sludy,
seven beagIe dogs vere foIIoved u for 3 years afler lhe induclion of rimary
hyolhyroidism. Three of lhese dogs vere foIIoved u for anolher 1.5 years vhiIe
receiving I-lhyroxine. AdenohyohyseaI funclion vas invesligaled al 2-monlh
inlervaIs vilh lhe combined inlravenous in|eclion of CRH, GHRH, GnRH, and
TRH, and measuremenl of lhe Iasma concenlralions of ACTH, GH, LH, IRL, and
TSH. In addilion, afler 2 years of hyolhyroidism a singIe TRH-slimuIalion lesl
and a somaloslalin lesl vere erformed, vilh measuremenls of lhe same iluilary
hormones. Ivery 6 monlhs lhe iluilary gIand vas visuaIized by comuled
lomograhy (CT).
Induclion of IH Ied lo high Iasma TSH concenlralions for a fev monlhs,
vhere afler concenlralions graduaIIy decIined lo vaIues no Ionger significanlIy
differenl from re-IH vaIues. A bIunled resonse lo slimuIalion of TSH reIease
receded lhis decIine. asaI Iasma GH concenlralions increased during IH and
lhere vas a aradoxicaI hyerresonsiveness lo TRH slimuIalion. asaI GH
concenlralions remained eIevaled and relurned onIy lo Iov vaIues during I-
lhyroxine lrealmenl. asaI IRL concenlralions decreased significanlIy during IH
and normaIized afler severaI monlhs of I-lhyroxine lrealmenl. The iluilary gIand
became enIarged in aII dogs. HislomorhoIogy and immunohislochemicaI sludies
in 4 dogs, afler 3 years of IH, reveaIed lhyrolroe hyerIasia, Iarge vacuoIaled
lhyroid deficiency ceIIs, and decreased numbers of mammolroes. SeveraI ceIIs
slained for bolh GH and TSH.
In concIusion, vilh lime IH Ied lo a Ioss of lhe TSH resonse lo Iov T4
concenlralions, hyersecrelion of GH, and hyosecrelion of IRL. The enIarged
iluilaries vere characlerized by lhyrolroe hyerIasia, Iarge vacuoIaled lhyroid
deficiency ceIIs, and doubIe-slaining ceIIs, vhich is indicalive of
lransdifferenlialion.



IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

117

Intrnductinn

Iiluilary ceIIs can adal by differenl mechanisms lo funclionaI demands
requiring hyersecrelion of a secific lrohic hormone, such as vhen a iluilary-
deendenl endocrine gIand is hyofunclionaI. Iirsl, decreased hormone
roduclion by a eriheraI gIand Ieads lo increased secrelion of lhe corresonding
iluilary hormone and increased numbers of secific iluilary ceIIs, according lo
lhe cIassic one-ceII-one-hormone concel. This concel reresenls lhe viev lhal
each adenohyohyseaI ceII lye roduces a singIe hormone lhal is secreled uon
slimuIalion by a arlicuIar hyolhaIamic-reIeasing hormone. Second, ceIIs of one
ceII Iine may be lransformed inlo anolher lo conlribule lo lhe demand for a secific
iluilary hormone. Thus, conlrary lo lhe reslriclive one-ceII-one-hormone concel,
adenohyohyseaI ceIIs are nol irrevocabIy monohormonaI bul may become
oIyhormonaI. The aIleralion of lhe morhoIogic fealures and lhe secrelory
caacily of malure ceII lyes vilhoul ceII division is caIIed lransdifferenlialion.
1, 2
The adenohyohyseaI changes associaled vilh insufficienl secrelion of a
eriheraI hormone have besl been sludied in exerimenlaI and cIinicaI
hyolhyroidism. In rals made hyolhyroid by adminislralion of royIlhiouraciI,
somalolroes lransform inlo slimuIaled lhyrolroes, so-caIIed lhyroid deficiency
ceIIs.
3
These lhyroid deficiency ceIIs are bihormonaI, in lhal lhey are
immunoreaclive for bolh GH and TSH. Thus, in addilion lo muIliIicalion of
lhyrolroes, lransdifferenlialion of somalolroes inlo lhyrosomalolroes
conlribules lo lhe increase in lhe number of TSH-roducing ceIIs.
4

In humans, rolracled rimary hyolhyroidism Ieads lo iluilary
enIargemenl due lo massive lhyrolroe hyerIasia and lhe formalion of lhyroid
deficiency ceIIs. The cIinicaI characlerizalion of lhe iluilary changes secondary lo
rimary hyolhyroidism is IargeIy based uon case sludies and consequenlIy is
somevhal fragmenlary. The usuaIIy asymlomalic enIargemenl of lhe iluilary
gIand may have lhe characlerislics of a macroadenoma.
5,6
Thyroid hormone
lheray commonIy Ieads lo regression of lhe iluilary mass,
6,7
bul refracloriness
has been reorled.
5
In hyolhyroid humans, lhe eIevaled basaI concenlralions of
bolh TSH and IRL resond in an exaggeraled manner lo TRH slimuIalion.
1,4
In
addilion, lhe Iasma GH concenlralions of alienls vilh hyolhyroidism resond
lo TRH, vhereas lhis resonse is absenl in normaI sub|ecls.
8
Hovever, nolhing is
knovn aboul lhe lime scaIe over vhich lhese changes deveIo.
In lhis conlexl, lhe dog deserves allenlion. Dogs can sonlaneousIy
deveIo rimary hyolhyroidism. RemarkabIy, in aboul one-lhird of cases basaI
Iasma TSH concenlralions are nol eIevaled,
9,10
aIbeil lhe uIsaliIily of TSH reIease
is more easiIy idenlified in hyolhyroid dogs lhan in heaIlhy dogs.
11
In dogs vilh
rimary hyolhyroidism, lhe TSH resonse afler adminislralion of TRH is
Chaler 7
_________________________________________________________________________________

118

generaIIy smaIIer lhan lhal in heaIlhy dogs and dogs vilh non-lhyroidaI iIIness.
12,13

The basaI Iasma GH concenlralions of hyolhyroid dogs are higher lhan lhose of
heaIlhy dogs,
14
and rimary hyolhyroid dogs resond lo TRH adminislralion
vilh a significanl rise in Iasma GH concenlralions, vhereas heaIlhy dogs do nol.
15
In order lo gain a beller underslanding of lhe deveIomenl of funclionaI
and morhoIogicaI changes in lhe iluilary gIand, ve have monilored
adenohyohyseaI funclion and iluilary size in dogs vilh induced rimary
hyolhyroidism for 4.5 years, before (3 years) and afler (1.5 years) lrealmenl vilh I-
lhyroxine.

Matcria!s and Mcthnds

Ocgs
Seven heaIlhy, neulered femaIe beagIe dogs vere enlered in lhe sludy al
lhe age of 3 years. Ovarieclomy had been erformed in aII dogs al lhe age of 12
monlhs. The dogs vere housed individuaIIy in indoor-ouldoor runs. They vere
fed on slandard manufaclured dry dog food lvice daiIy and given valer a! |i|iiun.
AII bilches vere accuslomed lo lhe Iaboralory environmenl and rocedures, such
as lhe coIIeclion of bIood.
Irimary hyolhyroidism vas induced by surgicaI removaI of lhe lhyroid
gIands, Ieaving lhe exlernaI aralhyroid gIands in silu. Aboul 3 monlhs Ialer, lhe
dogs vere fed on a Iov-iodine diel of meal and vhile bread for 4 veeks, vhich
decreased lhe urinary iodine conlenl from an average (+ SD) of 13.32 + 3.70 moI/I
lo 0.37 + 0.01 moI/I.
16
This vas done lo increase iodide ulake by ossibIy
remaining of lhyroid lissue, vhich vas lhen deslroyed by adminislering 5 mCi
(185 Mq) of sodium
131
iodide oraIIy.
The dogs vere fasled for 12 hours before lhe funclion lesls vhich vere aII
slarled belveen 8.00 and 10.00 hours. In lhe lhree dogs in vhich lhyroxine
subslilulion lheray vas slarled afler 3 years, medicalion vas omilled on lhe
morning of lhe lesls.

|unciicn icsis
Anlerior iluilary funclion vas invesligaled al 2-monlh inlervaIs using
combined adminislralion of four hyolhaIamic reIeasing hormones (lhe 4RH lesl):
CRH, GHRH, GnRH, and TRH vilh measuremenl of ACTH, GH, LH, IRL, and
TSH.
17,18

To lhis end, an inlravenous calheler vas Iaced in lhe cehaIic vein of each
dog lo faciIilale raid sequenliaI in|eclions. ImmedialeIy afler coIIeclion of lhe zero
bIood samIe from lhe |uguIar vein, lhe four hyolhaIamic reIeasing hormones
vere in|ecled inlravenousIy vilhin 30 seconds. The doses of lhe reIeasing
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

119

hormones used vere 1 g oCRH (IeninsuIa Laboralories Inc., eImonl, CA, USA)
er kg body veighl, 1 g hGHRH (IeninsuIa Laboralories Inc.) er kg body
veighl, 10 g GnRH (IerlagyI, Inlervel, oxmeer, NL) er kg body veighl, and
10 g TRH (TRH UC, IrolireIinum, S.A. UC N.V., Secleur Iharma, raine-
IAIIeud, eIgium) er kg body veighl. Iood samIes for lhe delerminalion of
Iasma concenlralions of ACTH, GH, LH, IRL, and TSH vere coIIecled from lhe
|uguIar vein al 15, 0, 5, 10, 20, 30, and 45 min, and immedialeIy Iaced in chiIIed
IDTA-coaled or hearinized (for TSH) lubes and cenlrifuged al 4C for 10 minules
al 3000 x g. IIasma vas slored al 25C unliI assayed.
Tvo years afler lhe induclion of hyolhyroidism a singIe TRH-slimuIalion
lesl vas erformed, using a TRH dose and rocedure idenlicaI lo lhose of lhe 4RH
lesl. A somaloslalin lesl vas carried oul vilh inlravenous in|eclion of 10 g er kg
body veighl of lhe somaloslalin anaIogue oclreolide (Sandoslalin, Novarlis
Iharma, Arnhem, NL). Iood coIIeclion and measuremenls vere as for lhe 4RH
lesl.
IIasma T4 and TSH concenlralions vere measured monlhIy in aII dogs
afler induclion of rimary hyolhyroidism during lhe lhree years of lhe sludy of
lhe hyolhyroid slalus.

Trcaincni
Three years afler lhe induclion of hyolhyroidism, lhe dogs vere
randomIy divided inlo lvo grous. Iour dogs vere eulhanized for
hislomorhoIogicaI sludies of lhe iluilary gIand, and lhree dogs received I-
lhyroxine (15 g/kg body veighl, oraIIy lvice daiIy).
19,20
In lhe Ialler dogs
delerminalion of Iasma T4 and TSH concenlralions, 4RH lesling, and iluilary
imaging vere conlinued al lhe same frequencies. Afler 1 year of reIacemenl
lheray vilh I-lhyroxine, lhese lhree dogs vere aIso eulhanized for
hislomorhoIogicaI sludies of lhe iluilary gIand.

Hcrncnc ncasurcncnis
IIasma lolaI T4 concenlralions vere delermined by a homoIogus soIid-
hase, chemiIuminescenl enzyme immunoassay (ImmuIile canine lolaI T4,
Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, CA, USA) in accordance
vilh lhe inslruclions of lhe manufaclurer and vaIidaled for lhe dog.
21
The inlra-
assay coefficienls of varialion (CVs) vere 13.8 and 8.2% al T4 IeveIs of 8 and 25
nmoI/I, resecliveIy. The inler-assay CV vas 8.5% al T4 IeveIs of 21 nmoI/I. The
sensilivily of lhe assay vas 2 nmoI/I.
IIasma TSH concenlralions vere delermined by a homoIogus soIid-hase,
lvo-siles chemiIuminescenl enzyme immunomelric assay (ImmuIile canine TSH,
Diagnoslic roducls (DIC), Los AngeIes, CA, USA), in accordance vilh lhe
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120

inslruclions of lhe manufaclurer. The inlra-assay CVs vere 5.0, 4.0, and 3.8% al
TSH IeveIs of 0.20, 0.50 and 2.6 g/I, resecliveIy. The inler-assay CVs vere 6.3 and
8.2% al TSH IeveIs of 0.16 and 2.8 g/I, resecliveIy. The sensilivily of lhe assay
vas 0.03 g/I. Cross-reaclivily vilh ISH and LH vas negIigibIe.
IIasma GH concenlralions vere delermined by a homoIogous RIA as
described reviousIy.
22
The inlra- and inler-assay CVs vere 3.8 and 7.2%,
resecliveIy. The sensilivily of lhe assay vas 0.3 g/I.
IIasma IRL concenlralions vere delermined vilh a reviousIy vaIidaled
heleroIogous RIA.
23
The inlra-assay and inler-assay CVs vere 3.5 and 11.5%,
resecliveIy. The sensilivily of lhe assay vas 0.8 g/I.
IIasma LH concenlralions vere delermined vilh a heleroIogous RIA
described earIier,
24
using a rabbil anliserum raised againsl ovine LH (CSU-204,
kindIy suIied by G D Nisvender, CoIorado Slale Universily, CO, USA), radio-
iodinaled bovine LH-7981 as reared for a bovine assay,
25
and canine iluilary
slandard LIR 1685-1 (a gifl of Dr L I Reicherl, AIbany MedicaI CoIIege, NY, USA).
The inlra-assay and inler-assay CVs for vaIues above 0.5 g/I vere 2.3% and 10.5%,
resecliveIy. The sensilivily of lhe assay vas 0.3 g/I.
IIasma ACTH concenlralions vere delermined vilh a commerciaIIy
avaiIabIe lvo-sile immunoradiomelric assay (IRMA) for humans (NichoIs Inslilule,
Wi|chen, NL). The inlra- and inler- assay CVs vere 3.2% and 7.8%, resecliveIy,
and lhe sensilivily vas 0.2 moI/I.

Ccnpuic! icncgrapnq
Comuled lomograhy (CT) of lhe iluilary gIand vas erformed every 6
monlhs. Iood vas vilhheId for 18 hours before imaging. IoIIoving lhe i.v.
adminislralion of 40-60 g medelomidine hydrochIoride (Domilor, SmilhKIine
eecham AnimaI HeaIlh V, Zoelermeer, NL) er kg body veighl, anaeslhesia vas
induced by i.v. adminislralion of 1-2 mg roofoI (Dirivan, Zeneca V,
Ridderkerk, NL) er kg body veighl and vas mainlained by inhaIalion of
haIolhane (HaIolhaan, AIbic V, MaassIuis, NL) or isofIuorane (Iorene, Abbol
Laboralories Lld, Queenborough, Kenl, UK), nilrous oxide, and oxygen.
Comuled lomograhy vas erformed vilh a lhird-generalion CT
scanner (Tomoscan CX/S, IhiIis NV, Iindhoven, NL).
26,27
Wilh lhe dog in slernaI
recumbency, lransverse scans of lhe skuII base vere made using a 9-second
scanning lime vilh 120 kV and 220mA. Conliguous 2-mm-lhick sIices vere made
from lhe roslraI cIinoid rocesses lo lhe dorsum seIIae, bolh before and afler lhe
inlravenous adminislralion of 2 mI of conlrasl medium er kg of body veighl
(TeIebrix 350, sodium and megIumine |oxilaIamale, conlaining 350 mg iodine/mI,
Guerbel NederIand V, Gorinchem, NL).

IunclionaI and morhoIogicaI changes in lhe adenohyohysis
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121

The heighl of lhe iluilary vas measured on lhe image lhal conlained lhe
Iargesl cross-seclion of lhe iluilary gIand in each examinalion for each dog.
28
AII
measuremenls vere erformed by one observer (R.H. vd V.-M.).

Hisicncrpnc|cgq an! innuncnisiccncnica| siu!ics
Afler eulhanasia, lhe skuII vas oened and lhe brain vas carefuIIy Iifled
from lhe skuII vilh lhe iluilary gIand allached lo lhe base of lhe diencehaIon.
The iluilary gIand vas immedialeIy removed vilh a surgicaI bIade and a median
sagillaI bIock vas Iaced in 10% neulraI buffered formaIin for 48 hours. The lissue
vas lhen roulineIy dehydraled and embedded in araffin. Seclions vere cul al 4-5
m for hemaloxyIin and eosin (HI) and immunohislochemicaI slaining.
SingIe immunohislochemicaI slaining vas erformed using lhe avidin-
biolin lechnique vilh a monocIonaI anlibody lo synlhelic ACTH1-24. Ior
immunohislochemicaI slaining of GH, TSH and IRL, rabbil oIycIonaI anlibodies
vere used againsl orcine TSH, orcine GH, and orcine IRL, resecliveIy. The
rimary anlibody characlerislics, manufaclurer, lye of anlibody and diIulion are
given in TabIe 1.
DoubIe-slaining for GH and TSH vas erformed on iluilary lissue from
lvo lhyroideclomized and lvo conlroI dogs (1.5-year-oId femaIe beagIe, 8-year-oId
neulered femaIe beagIe). Seclions vere roulineIy dearaffinized. Indogenous
eroxidase aclivily vas inhibiled vilh 1% H2O2 in melhanoI for 30 minules. The
seclions vere rinsed lhree limes, 5 minules each, vilh IS/Tveen-20. Non-secific
anlibody binding vas bIocked by incubalion vilh normaI goal serum (NGS, 1:10)
in IS for 30 minules. Seclions vere lhen incubaled for 60 minules vilh rabbil anli
GH 1:5000, rinsed vilh IS/Tveen-20 lhree limes, for 5 minules each, and
incubaled for 30 minules vilh goal anli-rabbil iolin IgG 1:250 in IS. Afler lhe
seclions vere rinsed vilh IS/Tveen-20 lhree limes, for 5 minules each, and lhen
incubaled overnighl vilh svine anli-rabbil IgGs 1:50 in IS and rinsed vilh
IS/Tveen-20 lhree limes, for 5 minules each. Then incubaled vilh Rabbil anli
TSH 1:3000 in IS al 4C. Afler lhe seclions vere vashed in IS/Tveen-20, lhree
limes, 5 minules each, lhey vere incubaled vilh Slrelavidin AI 1:50 in IS
during 45 minules and rinsed vilh IS lhree limes, for 5 minules each. The
seclions vere lhen equiIibraled in Tris HCI buffer (100 mM, H 8.2), 0.1% Tveen
20 for 10 minules, and incubaled in freshIy made veclor Iue Subslrale for 15
minules, foIIoved by vashing vilh Tris-HCI buffer (100 mM, H 8.2), 0.1% Tveen
20 for 5 minules, and IS lvo limes, for 5 minules each. Aflervards, lhey vere
incubaled vilh Invision, anli Rabbil for 45 minules. The seclions vere vashed
vilh IS lhree limes, for 5 minules each lime, incubaled in Veclor Nova Red for 10
minules, and vashed in MiIIi Q valer lhree limes, 5 minules each lime. They vere

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122

lhen dehydraled in series of aIcohoI 70%, 96% and 100% resecliveIy,
HISTOCLIAR (each 2 limes) for 5 minules each sle, and mounled in VeclaMounl.




Antibody Manufacturer Source Type Dilution
ACTH
Dept Inf Dis & Immunol Utrecht Univ MM 1:100
GH
Biogenesis 4750-3959 RP 1:5000
TSH
Biogenesis 8922-9989 RP 1:2000
PRL
Dept Farm Anim Health Utrecht Univ RP 1:8000

Tab!c 1. Source, lye of anlibody, and diIulion of anlibodies used for immunohislochemicaI
slaining. MM: mouse monocIonaI, RI: rabbil oIicIonaI, De Inf Dis & ImmunoI:
Dearlmenl of Infeclious Disease and ImmunoIogy, De Iarm Anim HeaIlh: Dearlmenl of
Iarm AnimaI HeaIlh, Ulrechl Univ: Ulrechl Universily.

Oaia ana|qsis
The area under lhe curve (AUC) above basaI hormone IeveIs vas
caIcuIaled using lhe lraezoidaI ruIe. ody veighls vere comared vilh Sludenl's
i-lesl for aired samIes. asaI hormone concenlralions, AUCs, and lhe heighls of
lhe iluilary gIands vere comared vilh lhe GeneraI Linear ModeI MuIlivariale
Tesl of SISS for Windovs (version 12.0.1, SISS Inc., Chicago, USA). Differences
vere considered significanl al Is0.05. Dala are resenled as means + SIM.

Ethics

The Ilhics Commillee of lhe IacuIly of Velerinary Medicine, Ulrechl
Universily, aroved lhe sludies.

IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

123

*

The Iasma T4 concenlralions of lhe seven dogs before lhyroideclomy (18.3
+ 1.7 nmoI/I) vere vilhin lhe reference range. Thyroideclomy and subsequenl
deslruclion of any remaining lhyroid lissue vilh
131
I induced lhe hysicaI changes
of hyolhyroidism: lhe dogs became Iess aclive and deveIoed lhick skin foIds and
aIoecia. The mean body veighl increased significanlIy (I0.002) from 14.5 + 0.7 kg
lo 20.1 + 1.4 kg. Iour dogs deveIoed maxiIIary rognalhia vilh videning of lhe
inlerdenlaI saces (Iig 1). IIasma T4 concenlralions measured monlhIy afler
induclion of rimary hyolhyroidism vere undeleclabIe (< 2 nmoI/I) during lhe
vhoIe sludy. In lhe lhree dogs in vhich reIacemenl lheray vilh I-lhyroxine vas
slarled lhree years afler lhyroideclomy, lhe characlerislics of hyolhyroidism
disaeared. They became more IiveIy, and a nev hair coal aeared. ody veighl
decreased vilh an average of 6 kg and lhe Iasma T4 concenlralions normaIized.

Basa| a!cncnqpcpnqsca| ncrncnc ccnccniraiicns
asaI Iasma TSH concenlralions increased afler lhyroideclomy, vilh lhe
highesl concenlralions being measured al 2 and 4 monlhs afler lhyroideclomy.
Thereafler Iasma TSH concenlralions decreased graduaIIy, aIlhough lhey
remained significanlIy higher lhan lhe iniliaI vaIues for mosl of lhe observalion
eriod. Hovever, al lhe end of lhis arl of lhe sludy, lhal is 3 years afler
lhyroideclomy, Iasma TSH concenlralions vere nol significanlIy differenl from
lhe concenlralions rior lo lhyroideclomy. Al 2.5 and 3 years afler lhyroideclomy
lhe Iasma TSH concenlralions vere vilhin lhe reference range in one and lvo
dogs, resecliveIy. Afler 3 years, six oul of lhe 7 dogs had Iasma TSH
concenlralions above lheir re-lhyroideclomy vaIues. Wilh lhyroxine subslilulion
lhe Iasma TSH concenlralion furlher decIined (Iig 2), oflen lo undeleclabIe
concenlralions.
asaI Iasma GH concenlralions increased graduaIIy during lhe firsl year
and remained al high IeveIs during lhe foIIoving 2 years (Iig 2). Subslilulion vilh
I-lhyroxine caused lhe Iasma GH concenlralion lo decIine lo lhe iniliaI Iov vaIues
vilhin 2 monlhs (Iig 2).
The basaI Iasma IRL concenlralions did nol change in lhe firsl 6 monlhs
afler lhyroideclomy bul decreased lhereafler, remaining Iov for lhe resl of lhe
sludy eriod (Iig 2). IIasma IRL concenlralions increased significanlIy afler 1 year
of lhyroxine subslilulion lheray (Iig 2).
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Figurc 2. Mean (+ SIM) basaI Iasma concenlralions of TSH, GH, IRL, LH and ACTH
measured al 2-monlhIy inlervaIs in seven beagIe dogs vilh hyolhyroidism induced al lime
oinl 0. Three of lhese dogs vere foIIoved u for anolher 1.5 years vhiIe on I-lhyroxine
subslilulion (beginning of subslilulion marked by arrov). Aslerisks indicale slalislicaIIy
significanl difference from vaIue al lime zero.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

125

asaI Iasma LH concenlralions increased afler lhe induclion of
hyolhyroidism and significanlIy (I0.04) 8-12 monlhs afler lhyroideclomy.
Thereafler, lhe basaI Iasma LH concenlralions decreased again and vere no
Ionger slalislicaIIy differenl from lhe iniliaI concenlralions (Iig 2). In lhe lhree dogs
in vhich I-lhyroxine subslilulion vas slarled afler lhree years of hyolhyroidism,
lhe basaI Iasma LH concenlralions lended lo rise (Iig 2).
asaI Iasma ACTH concenlralions remained raclicaIIy unchanged, vilh
lhe excelion of lhe vaIues al 4 and 20 monlhs afler lhe induclion of
hyolhyroidism. Al lhese lime oinls lhe Iasma ACTH concenlralions vere
significanlIy higher (I0.03) and Iover (I0.03), resecliveIy, lhan lhe iniliaI
concenlralions (Iig 2). Trealmenl vilh I-lhyroxine had no effecl on lhe basaI
Iasma ACTH concenlralions (Iig 2).


In lhe 4RH lesl, slimuIaled TSH reIease, caIcuIaled as lhe AUC above basaI
vaIues, did nol change during lhe firsl 2 monlhs afler lhyroideclomy, bul
decreased sharIy lhereafler. The AUC above baseIine slayed Iov for lhe resl of
lhe sludy, vilh mosl vaIues being significanlIy Iover lhan lhe iniliaI vaIues (Iig 3).
One year afler lhyroxine subslilulion a lendency lo reslored resonsiveness vas
noled (Iig 3).
AIong vilh lhe increasing basaI Iasma GH concenlralions, lhe GH reIease
uon slimuIalion increased as veII, aIbeil vilhoul reaching slalislicaI significance
(Iig 3). In lhe lhird year of hyolhyroidism, lhe AUC for GH decreased and a
furlher decrease occurred in lhe lhree dogs on I-lhyroxine subslilulion (Iig 3).
UnliI 10 monlhs afler lhyroideclomy lhe AUCs for IRL vere significanlIy
increased (I<0.05). Irom 6 monlhs onvards, lhe resonsiveness in lhe 4RH lesl
graduaIIy decreased reaching significanlIy Iover IRL resonsiveness lhan in lhe
eulhyroid slale al lhe end of lhe 3-y eriod (Iig 3). IRL resonsiveness vas nol
reslored by I-lhyroxine subslilulion.
The slimuIaled Iasma LH concenlralions increased vilh lime, reaching
slalislicaI significance al 6 monlhs afler induclion of hyolhyroidism. The
slimuIaled Iasma LH concenlralions did nol change significanlIy vilh lime in lhe
lhree dogs in vhich I-lhyroxine subslilulion vas begun afler lhree years of
hyolhyroidism (Iig 3).
The ACTH reIease uon slimuIalion vas nol significanlIy differenl from
lhal before lhyroideclomy and vas nol affecled by I-lhyroxine subslilulion (Iig 3).

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Figurc 3. Mean (+ SIM) AUCs above baseIine of lhe slimuIaled Iasma concenlralions of
TSH, GH, IRL, LH and ACTH in seven hyolhyroid beagIe dogs (see aIso Iegend lo Iig 2).
SlimuIalion comrised lhe raid inlravenous in|eclion of 4 hyolhaIamic reIeasing
hormones (4-RH lesl) al 2-monlhIy inlervaIs.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

127

T|H-an! scnaicsiaiin icsi
Tvo years afler lhyroideclomy, basaI TSH IeveIs exceeded lhe uer Iimil
of lhe reference range (0.6 g/I), bul lhere vas nol a significanl resonse lo TRH
slimuIalion (Iig 4), as couId be execled from lhe TSH resonse in lhe 4RH lesl al
lhis oinl in lime (Iig 3). Hovever, lhe TRH slimuIalion lesl resuIled in a
significanl increase in Iasma GH concenlralions al 30 and 45 minules (I0.013 and
I0.002, resecliveIy) (Iig 4) and in non-significanl changes in IRL, ACTH and LH
concenlralions.
IIasma concenlralions of TSH, GH, IRL, LH, and ACTH did nol change
significanlIy afler lhe adminislralion of somaloslalin (nol shovn).

Ccnpuic! icncgrapnq cj inc piiuiiarq g|an!
The iluilary gIand sIovIy enIarged afler lhe induclion of hyolhyroidism
(Iig 5). efore lhyroideclomy lhe iluilary heighl ranged from 3.5 lo 4.1 mm (mean
3.8 mm), and al 24 monlhs afler lhe induclion of hyolhyroidism il ranged from
4.8 lo 5.5 mm (mean 5.1 mm). In lhe lhree dogs receiving reIacemenl lheray lhe
iluilary gIand subsequenlIy decreased in size. ReIacemenl lheray Ied lo
normaIizalion of iluilary size afler 15 monlhs (Iig 6).






















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Figurc 4. IIasma concenlralions of TSH, GH, IRL, LH and ACTH afler singIe inlravenous
in|eclion of TRH in seven beagIes dogs afler 2 years of hyolhyroidism.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

129

































Figurc 5. Transverse CT images of lhe skuII of a beagIe rior lo lhyroideclomy (Iefl), and 3
years afler lhyroideclomy (righl). Conlrasl enhancemenl visuaIized lhe normaI-size
iluilary gIand before lhyroideclomy and lhe enIargemenl afler induclion of
hyolhyroidism.




IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

129

































Figurc 5. Transverse CT images of lhe skuII of a beagIe rior lo lhyroideclomy (Iefl), and 3
years afler lhyroideclomy (righl). Conlrasl enhancemenl visuaIized lhe normaI-size
iluilary gIand before lhyroideclomy and lhe enIargemenl afler induclion of
hyolhyroidism.




204988_p129cc.indd 1 11-11-2008 13:32:40
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130
































Figurc 6. Mean (+ SIM) iluilary heighls measured on CT images made al 6-monlhIy
inlervaIs in seven beagIes vilh hyolhyroidism induced al lime oinl zero. Afler 3 years of
hyolhyroidism in lhree of lhese dogs I-lhyroxine subslilulion lheray vas slarled (arrov),
resuIling in a decrease in iluilary heighls lovard re-lhyroideclomy vaIues.
SignificanlIy differenl from iniliaI heighl.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

131

Hisicncrpnc|cgq
Mosl adenohyohyseaI ceIIs from lhe hyolhyroid dogs slained
acidohiIic. These acidohiIic ceIIs had a normaI size (7-9 m) or vere enIarged
(10-19 m) vilh Iarge nucIei (sized 5 m). Many very Iarge aIe ceIIs (15-32 m)
vere observed lhroughoul lhe adenohyohysis, oflen vilh vacuoIar degeneralion
of lhe cyloIasm, a Iarge round nucIeus (8-12 m) vilh a rominenl nucIeoIus, and
somelimes doubIe nucIei (Iig 7-1). SmaII chromohobe and basohiIic ceIIs of
normaI size (7-9 m) vere aIso seen. The adenohyohyses of lhe hyolhyroid
dogs lrealed vilh I-lhyroxine conlained acidohiIic ceIIs and chromohobe and
basohiIic ceIIs (7-9 m) simiIar lo lhose found in heaIlhy dogs, as veII as some
smaII aggregales of Iarger basohiIic ceIIs. No Iarge vacuoIaled ceIIs, as observed in
lhe hyolhyroid dogs, vere seen.
TSH-immunoosilive ceIIs vere slrongIy increased in number in lhe
hyolhyroid dogs as comared vilh lhe conlroI dogs. Many of lhe Iarge
vacuoIaled ceIIs slained osilive for TSH (Iig 7-2). Among lhe many GH-
immunoosilive ceIIs lhere vere aIso some Iarge vacuoIaled ceIIs (Iig 8-1). DoubIe-
immunoIabeIing (Iig 8-2) reveaIed lhal lhere vere GH-osilive ceIIs (bIue) and
TSH-osilive ceIIs (orange). SeveraI, arlicuIarIy smaII, ceIIs shoved doubIe
slaining for GH and TSH (brovn-bIue). This doubIe slaining for GH and TSH vas
aIso found in normaI-sized ceIIs in heaIlhy conlroI dogs, aIbeil in a much Iover
frequency.
IRL immunoosilive ceIIs vere smaII and cIearIy diminished in number in
lhe hyolhyroid dogs, as comared vilh lhe conlroI dogs. AIso some of lhe Iarge
vacuoIaled ceIIs slained osilive for IRL (Iig 9-1). There vere many more IRL-
immunoosilive ceIIs in lhe I-lhyroxine lrealed dogs lhan in lhe hyolhyroid dogs.
In lhe subsliluled dogs lhere vere bolh smaII and Iarge IRL immunoosilive ceIIs,
lhe Ialler oflen in smaII aggregales (Iig 9-2)



As execled, lhyroideclomy induced lhe hysicaI characlerislics of
hyolhyroidism. Hovever, as reorled earIier, lhere vere aIso signs of
acromegaIy.
14
In rinciIe, lhe hyolhyroidism-induced GH excess couId be of
exlra-iluilary origin. In dogs rogeslerone may induce excessive GH secrelion,
29

originaling from foci of hyerIaslic ducluIar eilheIium of lhe mammary
gIand.
30,31,32
This mammary GH is reIeased evenIy lhroughoul lhe day, vhereas in
dogs vilh rimary hyolhyroidism lhe eIevaled GH secrelion sliII has a uIsaliIe
nalure, even lhough uIsaliIe GH reIease is smaIIer lhan lhal in heaIlhy dogs.
14

Moreover, lhe ovarieclomized slalus of lhe hyolhyroid dogs makes lhe
occurrence of rogeslerone-induced GH excess very unIikeIy. So far, lhis co-
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occurrence of rimary hyolhyroidism and acromegaIy seems lo be unique lo
dogs. Il has nol been reorled in olher animaIs vilh exerimenlaIIy induced
hyolhyroidism or in humans vilh iluilary hyerIasia secondary lo rimary
hyolhyroidism.
5

In Iine vilh lhe cIassicaI endocrine rinciIes of feedback reguIalion, lhe
induclion of rimary hyolhyroidism caused lhe Iasma TSH concenlralions lo
rise by a faclor of aboul 15. Hovever, lhe Iasma TSH concenlralions remained al
lhis high IeveI for onIy a fev monlhs and lhen graduaIIy decIined lo reach
concenlralions nol significanlIy differenl from lhe iniliaI IeveIs. The resonse lo
slimuIalion decreased even fasler. These observalions can exIain lhe oflen non-
eIevaled basaI Iasma TSH concenlralions and lhe oflen bIunled or absenl TSH
resonse lo TRH slimuIalion in dogs vilh sonlaneous rimary
hyolhyroidism.
9,10,12,13
In conlrasl, case reorls of humans vilh Iong-slanding
rimary hyolhyroidism indicale lhal basaI circuIaling TSH concenlralions are
high and lhal lhese alienls resond lo TRH slimuIalion in an exaggeraled
fashion.
6
In humans a bIunled TSH resonse lo slimuIalion vilh TRH seems lo
occur onIy in alienls vilh TSH-secreling iluilary lumors.
33

In conlrasl lo lhe basaI TSH concenlralions lhe basaI GH concenlralions
did nol decIine vilh lime and onIy relurned lo re-lhyroideclomy vaIues vilh I-
lhyroxine lrealmenl. In olher secies lhe silualion seems differenl, as basaI GH
concenlralions have nol been reorled lo be high in hyolhyroidism. There is
evidence lhal in humans and rals hyolhyroidism is associaled vilh a slrong
decrease in lhe number of GH-immunoosilive ceIIs.
2,4
Hovever, aIlhough in olher
secies hyolhyroidism may nol be associaled vilh hyersomalolroism, lhe
resonse lo slimuIalion aears lo be simiIar lo lhal in dogs. In lhe resenl sludy,
lhe GH resonses lo slimuIalion in lhe 4RH lesl varied slrongIy and seemed lo
increase and decrease vilh lime. Of lhe hormones adminislered, nol onIy GHRH
bul aIso TRH viII have increased lhe GH reIease. In lhis sludy and in an earIier
sludy ve found lhal hyolhyroid dogs have a aradoxicaI GH resonse lo singIe
TRH adminislralion.
15
A simiIar aberranl GH resonse lo TRH has been reorled
in humans.
8

The eIevaled Iasma GH concenlralions did nol resond lo somaloslalin
adminislralion. Lack of suression of basaI Iasma GH concenlralions by
oclreolide has been reorled in heaIlhy dogs,
34,35
hovever, oclreolide relrealmenl
comIeleIy bIunls lhe GH resonse in dogs slimuIaled vilh GHRH.
35
In humans
oclreolide rimariIy decreases lhe amounl of circuIaling GH by suressing lhe
mass of GH reIeased er secrelory bursl, aIlhough lhe inleruIse (lrough) IeveIs are
aIso Iovered.
36
In hyolhyroid dogs GH is sliII secreled in uIses,
14
bul lhe effecl of
oclreolide on uIsaliIe GH reIease has nol yel been sludied.

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133

One of lhe resenling fealures of (rolracled) rimary hyolhyroidism in
humans is gaIaclorrhea.
37
Oflen lhere is aIso hyerroIaclinemia.
4,7,37,38

GaIaclorrhea is nol a common fealure of canine hyolhyroidism. Il vas aIso nol
observed in lhe resenl beagIes and lheir basaI Iasma IRL concenlralions did nol
increase afler induclion of hyolhyroidism. In facl, Iasma IRL concenlralions
decreased significanlIy aboul 6 monlhs afler lhyroideclomy. SimiIarIy, lhe iniliaIIy
increased resonsiveness in lhe 4RH lesl decreased vilh lime, vilh evenluaIIy a
significanlIy decreased resonsiveness. OnIy afler severaI monlhs of I-lhyroxine
reIacemenl lheray did basaI Iasma IRL concenlralions increase lovard re-
lhyroideclomy IeveIs.
Ovarieclomy in dogs increases lhe basaI Iasma LH concenlralions by a
faclor of aboul 30.
39
In lhe currenl sludy, lhe high basaI Iasma LH concenlralions
resonded iniliaIIy lo lhe induclion of hyolhyroidism vilh a modesl increase bul
remained raclicaIIy lhe same lhroughoul lhe sludy. L-lhyroxine suIemenlalion
did nol significanlIy change LH concenlralions. AarenlIy, hyolhyroidism does
nol decrease Iasma LH concenlralions - as has been observed in rals, humans and
monkeys in ovarieclomized beagIes.
40,41,42

The induclion of hyolhyroidism caused iluilary enIargemenl in aII dogs,
vhereas iluilary hyerIasia secondary lo rimary hyolhyroidism aears lo be
rare in humans. Hovever, a recenl series of alienls vilh rimary hyolhyroidism
reveaIed iluilary enIargemenl on MRI in 70% of cases.
37
L-lhyroxine
adminislralion Ied lo regression of lhe iluilary enIargemenl in lhe lhree lrealed
dogs, as has been reorled in some humans vilh iluilary hyerIasia secondary
lo rimary hyolhyroidism.
6,7,43

Taken logelher, vilh lime hyolhyroidism caused: (1) lhe iniliaIIy eIevaled
Iasma TSH concenlralions lo normaIize vilh Ioss of resonse lo slimuIalion, (2)
Iasma GH concenlralions lo rise lo abnormaIIy high IeveIs vilh a aradoxicaI
resonse lo TRH slimuIalion, and (3) Iasma IRL concenlralions lo decrease.
Changes in Iasma LH and ACTH concenlralions vere essenliaIIy unremarkabIe.
The funclionaI changes vere associaled vilh sIovIy rogressing iluilary
enIargemenl, being reversibIe on lrealmenl vilh I-lhyroxine.
Iiluilary hislomorhoIogy shoved massive lhyrolroe hyerIasia in lhe
unlrealed hyolhyroid dogs. The Iarge, aIe ceIIs had abundanlIy vacuoIaled
cyloIasm, simiIar lo lhe so-caIIed lhyroideclomy or lhyroid deficiency ceIIs in
hyolhyroid rals and in humans vilh unlrealed rolracled rimary
hyolhyroidism.
3,4
We suggesl lhal lhe ersislenl slimuIalion of lhyrolroes via
lhe negalive feedback Ioo uIlimaleIy caused TRH-recelor desensilizalion,
Ieading lo graduaIIy decreasing TSH reIease and Ioss of resonsiveness lo
slimuIalion. Raid desensilizalion eIiciled by a Iov concenlralion of agonisl

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134

allenuales hysioIogicaI ceIIuIar resonses, vhereas ersislenl desensilizalion
eIiminales lhis reguIalory mechanism.
44,45

The resence of miloses in lhyrolroe ceIIs suggesls lhal lhe division of
re-exisling lhyrolroes and/or Iess differenlialed (slem) ceIIs conlribuled lo lhe
deveIomenl of lhe lhyroid deficiency ceIIs. In addilion lo lhis mechanism, lhe
resuIls of doubIe immunoslaining indicale lhal anolher mechanism aIso vas
oeralionaI. SeveraI smaII ceIIs immunoslained for bolh TSH and GH, simiIar lo
lhe changes lhal have been reorled in hyolhyroid rals and humans.
3,4
This
lransdifferenlalion of somalolroes inlo lhyrolroes may aIso exIain lhe
aradoxicaI GH secrelion uon slimuIalion vilh TRH observed in dogs vilh
sonlaneous rimary hyolhyroidism.
15
The recenlIy idenlified muIlifunclionaI
ceIIs lhal slore and secrele lvo differenl adenohyohyseaI hormones
(oIyhormonaI ceIIs) and/or resond lo severaI reIeasing hormones may be
invoIved in lransdifferenlialion and aradoxicaI secrelion.
46
We aIso found a fev
adenohyohyseaI ceIIs lhal slained for bolh TSH and GH in conlroI dogs.
Wilh lime lhe GH resonse lo slimuIalion seemed lo decrease, aIlhough
Ialer and Iess ronounced lhan lhe TSH resonse. In conlrasl lo basaI TSH
concenlralions, basaI GH concenlralions remained high. Thus il vouId seem lhal
lhe roosed exIanalion (ermanenl desensilizalion) for lhe Iov resonsiveness
of TSH lo TRH does nol hoId lrue for GH reIease. One exIanalion is lhal lhe TRH
recelors exressed on lhyrosomalolroes are Iess rone lo desensilizalion lhan
lhe TRH recelors on lhe (monohormonaI) lhyrolroes. The amounl of TSH lhal
mighl be secreled by lhese lhyrosomalolroes afler TRH-slimuIalion is obviousIy
insufficienl lo resuIl in enhanced Iasma TSH concenlralions.
The observed decrease in basaI IRL concenlralions and lhe decrease in
IRL resonsiveness lo slimuIalion vilh lime may aIso be a resuIl of
lransdifferenlialion. In humans, Iong-slanding acromegaIy due lo ecloic
overroduclion of GHRH is associaled vilh somalolroe hyerIasia combined
vilh lhe aearance of mammosomalolroes, derived from Iaclolroes.
1
The
number of monohormonaI Iaclolroes vas very Iov in lhese alienls. Againsl lhe
background of lhese observalions, lhe graduaI decrease in Iaclolroe aclivily in our
dogs couId be due lo recruilmenl of Iaclolroes lo form mammosomalolroes and
ossibIy lhyromammosomalolroes.
In concIusion, vilh lime lhe abrul induclion of rimary hyolhyroidism
Ied lo severaI funclionaI and morhoIogicaI changes in lhe adenohyohysis. The
main funclionaI changes vere graduaI Ioss of TSH resonse lo lhe Iov circuIaling
T4 concenlralions beginning afler 4 monlhs, graduaI deveIomenl of
hyersecrelion of GH in lhe firsl year vhich lhen remained high, and
hyosecrelion of IRL afler 6 monlhs. HislomorhoIogicaIIy, lhe enIarged
iluilaries vere characlerized by lhyrolroe hyerIasia, Iarge vacuoIaled lhyroid
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

135

deficiency ceIIs, and doubIe-slaining ceIIs, indicalive of lransdifferenlialion. These
observalions exIain severaI of lhe earIier observed biochemicaI fealures of canine
hyolhyroidism, aIlhough lhe resenl findings may nol be fuIIy reresenlalive of
lhe graduaI sonlaneous disease in olher breeds, ages and gender.

Acknnw!cdgmcnts

Urinary lolaI iodine conlenls vere measured in lhe Iaboralory of lhe
Division of Iedialric IndocrinoIogy, Imma ChiIdren's HosilaI, Academic
MedicaI Cenler, Universily of Amslerdam, NL. The aulhors gralefuIIy
acknovIedge lhe heI of Irof. Dr. }.}.M. de Vi|Ider and lhe Iaboralory slaff. The
heI of Dr. }. van den roek in lhe slalislicaI anaIyses is highIy arecialed. The
aulhors are very gralefuI for lhe assislance of Mr. H.G.H. van IngeIen and Mrs.
Y.W.I.A. IoIIak.


























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Figurc 1. 5-year-oId beagIe vilh fealures of hyolhyroidism: lragic faciaI exression and
dorsaI nasaI aIoecia vilh igmenlalion. This beagIe aIso deveIoed maxiIIary rognalhia
vilh videning of lhe inlerdenlaI saces.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

137


















Figurc 7-1


















Figurc 7-2
Figurc 7. Seclions of lhe iluilary of a dog vilh rimary hyolhyroidism slained vilh HI (7.1) and an
anlibody againsl TSH (7.2). Many Iarge aIe ceIIs, oflen vilh vacuoIaled cyloIasm, a Iarge round
nucIeus, and occasionaIIy doubIe nucIei. The number of TSH-immunoosilive ceIIs is slrongIy increased
in comarison vilh lhe number in heaIlhy dogs.
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138



















Figurc 8-1


















Figurc 8-2
Figurc 8. Seclions of lhe iluilary gIand of a hyolhyroid dog slained vilh an anlibody againsl GH
(brovn, 8.1), and doubIe slained (8.2) for GH (bIue) and TSH (orange). olh Iarge and normaIIy sized
ceIIs slain osilive for GH. SeveraI ceIIs lhal slain osilive for bolh GH and TSH.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
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139


















Figurc 9-1


















Figurc 9-2
Figurc 9. Seclions of lhe iluilary of a hyolhyroid dog slained vilh an anlibody againsl IRL.
Comared vilh heaIlhy conlroI dogs lhere vere onIy fev immunoosilive ceIIs (9.1), incIuding some of
lhe Iarge vacuoIaled ceIIs. Trealmenl vilh I-lhyroxine Ied lo a slrong increase in lhe number of IRL-
immunoosilive ceIIs (9.2)
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140

*

1. VidaI S, Horvalh I, Kovacs K, LIoyd RV, Smylh HS 2001 ReversibIe lransdifferenlialion:
Inlerconversion of somalolrohs and Iaclolrohs in iluilary hyerIasia. Mod IalhoI
14:20-28.
2. Radian S, CocuIescu M, Morris }I 2003 Somalolroh lo lhyrolroh ceII lransdifferenalialion
during exerimenlaI hyolhyroidism a Iighl and eIeclron-microscoy sludy. } CeII MoI
Med 7:297-306.
3. Horvalh I, LIoyd RV, Kovacs K 1990 IroyIlhiouracyI-induced hyolhyroidism resuIls in
reversibIe lransdifferenlialion fo somalolrohs inlo lhyroideclomy ceIIs. A morhoIogic
sludy of lhe ral iluilary incIuding immunoeIeclron microscoy. Lab Invesl 63:511-520.
4. VidaI S, Horvalh I, Kovacs K, Cohen SM, LIoyd RV, Scheilhauer W 2000
Transdifferenlialion of somalolrohs lo lhyrolrohs in lhe iluilary of alienls vilh
rolracled rimary hyolhyroidism. Virchovs Arch 436:43-51.
5. }oshi AS, WooIf ID 2005 Iiluilary hyerIasia secondary lo rimary hyolhyroidism: A
case reorl and a reviev of lhe Iileralure. Iiluilary 8:99-103.
6. SarIis N}, ruckner-Davis I, Doman }L, SkaruIis MC 1997 MRI-demonslrabIe regression
of a iluilary mass in a case of rimary hyolhyroidism afler a veek of acule lhyroid
hormone lheray. } CIin IndocrinoI Melab 82:808-811.
7. VaIenla L}, Tamkin }, Soslrin R, IIias AN, Iisenberg H 1983 Regression of a iluilary
adenoma foIIoving Ievolhyroxine lheray of rimary hyolhyroidism. IerliI SleriI 40:389-
392.
8. Hanev K, Ulsumi A, Sugavara A, Tanaka A, Iukazava H, Yoshida K, Abe K 1995
Inhanced Iasma GH resonses lo simuIlaneous adminislralion of TRH and GHRH in
alienls vilh rimary hyolhyroidism. Indocr } 42:43-47.
9. Ielerson MI, MeIian C, NichoIs R 1997 Measuremenls of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroin concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Ass 211:1396-1402.
10. orelli IS, reyer-Haube I, Kasers , Reusch CI . KIinische, hmaloIogische,
biochemische und endokrinoIogische Asekle bei 32 Hunden mil Hyolhyreose. |CIinicaI,
hemaloIogicaI, biochemicaI and endocrinoIogicaI asecls of dogs vilh hyolhyroidismj
Schveiz Arch TierheiIkd 2003,145:149-158.
11. Kooislra HS, Diaz-Isineira M, MoI }A, van den rom WI, Ri|nberk A 2000 Secrelion
allern of lhyroid-slimuIaling hormone in dogs during eulhyroidism and hyolhyroidism.
Domesl Anim IndocrinoI 18:19-29.
12. Scoll-Moncrieff }CR, NeIson RW 1998 Change in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolroin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and eulhyroid dogs vilh concurrenl disease. } Am Vel Med Ass
213:1435-1438.

IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

141

13. Diaz Isineira MM, MoI }A, Ieelers MI, IoIIak YWIA, Iversen L, van Di|k }I, Ri|nberk A,
Kooislra HS 2007 Assessmenl of lhyroid funclion in dogs vilh Iov Iasma lhyroxine
concenlralion. } Vel Inl Med 21:25-32.
14. Lee WM, Diaz-Isineira M, MoI }A, Ri|nberk A, Kooislra HS 2001 Irimary hyolhyroidism
in dogs is associaled vilh eIevaled GH reIease. } IndocrinoI 168:59-66.
15. Diaz-Isineira MM, GaIac S, MoI }A, Ri|nberk A, Kooislra HS 2008 Thyrolroin-reIeasing
hormone-induced grovlh hormone secrelion in dogs vilh rimary hyolhyroidism.
Domesl Anim IndocrinoI 34: 176-181.
16. Gons MH, Kok }H, TegeIaers WHH, de Vi|Ider }}M 1983 Concenlralion of Iasma
lhyrogIobuIin and urinary excrelion of iodinaled maleriaI in lhe diagnosis of lhyroid
disorders in congenilaI hyolhyroidism. Acla IndocrinoI 104:27-34.
17. Mei| I, MoI }A, HazevinkeI HAW, evers MM, Ri|nberk A 1996 Assessmenl of a
combined anlerior iluilary funclion lesl in beagIe dogs: raid sequenliaI inlravenous
adminislralion of four hyolhaIamic reIeasing hormones. Domesl Anim IndocrinoI 13:161-
170.
18. Mei| I, MoI }A, Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13:465-468.
19. IeIdman IC, NeIson RW 1996 Hyolhyroidism. In: Canine and IeIine IndocrinoIogy and
Reroduclion. 2
nd
edilion. IeIdman IC and NeIson RW (Ids.). IhiIadeIhia, IA: W
Saunders Comany. :55-85.
20. Ri|nberk A 1996 Thyroids. In: CIinicaI IndocrinoIogy of Dogs and Cals. 1
sl
edilion. Ri|nberk
A (Id.). Dordrechl: KIuver Academic IubIishers. :35-59.
21. runer }M, Scoll-Moncrieff CR, WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. }
Am Vel Med Assoc 212:1572-1575.
22. Iigenmann }I, Iigenmann RY 1981 Radioimmunoassay of canine grovlh hormone. Acla
IndocrinoI 98:514-520.
23. Okkens AC, evers MM, DieIeman S}, WiIIemse AH 1985 Shorlening of lhe inleroeslrous
inlervaI and lhe Iifesan of lhe corus Iuleum of lhe cycIic dog by bromocriline lrealmenl.
Vel Quarl 7:173-176.
24. Nell TM, Akbar AM, Ihemisler RD, HoIsl IA, RIiherl LI }r, Nisvender GD 1975 LeveIs of
Iuleinizing hormone, eslradioI and rogeslerone in serum during lhe eslrous cycIe and
regnancy in lhe beagIe bilch. Iroc Soc Ix ioI Med 148:134-139.
25. DieIeman S}, evers MM 1987 Iffecls of monocIonaI anlibody againsl IMSG adminislered
shorlIy afler lhe reovuIalory LH surge on lime and number of ovuIalions in IMSG/IG-
lrealed covs. } Rerod IerliI 81:533-542.
26. Van der VIugl-Mei|er RH, Mei| I, Voorhoul G 2006 Inlraobserver and inlerobserver
agreemenl, reroducibiIily, and accuracy of comuled lomograhic measuremenls of
iluilary gIand dimensions in heaIlhy dogs. Am } Vel Res 67:1750-1755.
Chaler 7
_________________________________________________________________________________

142

27. Van der VIugl-Mei|er RH, Voorhoul G, Mei| I 2002 Imaging of lhe iluilary gIand in dogs
vilh iluilary-deendenl hyeradrenocorlicism. MoI CeII IndocrinoI 197: 81-87.
28. Kooislra HS, Voorhoul G, MoI }A, Ri|nberk A 1997 CorreIalion belveen imairmenl of
gIucocorlicoid feedback and lhe size of lhe iluilary gIand in dogs vilh iluilary-
deendenl hyeradrenocorlicism . } IndocrinoI 152:387-394.
29. Iigenmann }I, Iigenmann RY, Ri|nberk A, van der Gaag I, Zaf }, Iroesch IR 1983
Irogeslerone-conlroIIed grovlh hormone overroduclion and naluraIIy occurring canine
diabeles and acromegaIy. Acla IndocrinoI 104:167-176.
30. SeIman I}, MoI }A, Rulleman GR, van Garderen I, Ri|nberk A 1994 Irogeslin-induced
grovlh hormone excess in lhe dog originales in lhe mammary gIand. IndocrinoIogy
134:287-292.
31. Van Garderen I, de Wil M, Voorhoul WI, Rulleman GR, MoI }A, Nederbragl H, Misdor
W 1997 Ixression of grovlh hormone in canine mammary lissue and mammary lumors.
Ividence for a olenliaI aulocrine/aracrine slimuIalory Ioo. Amer } Ialh 150:1037-1047.
32. halli SIM, Tao NAS, Okkens AC, MoI }A, Duchaleau L, DucaleIIe R, van den Ingh
TSGAM, TshamaIa M, Van Ham LML, Coryn M, Ri|nberk A, Kooislra HS 2007 RoIe of
rogeslin-induced mammary-derived grovlh hormone in lhe alhogenesis of cyslic
endomelriaI hyerIasia in lhe bilch. Domesl Anim IndocrinoI 33:294-312.
33. rucker-Davis I, OIdfieId IH, SkaruIis MC, Doman }L Weinlraub D 1999 Thyrolroin-
secreling iluilary lumors: Diagnoslic crileria, lhyroid hormone sensilivily, and lrealmenl
oulcome in 25 alienls foIIoved al lhe NalionaI Inslilules of HeaIlh. } CIin IndocrinoI
Melab 84:476-486.
34. Robben }H, van den rom WI, MoI }A, van Haeflen TW, Ri|nberk A 2006 Iffecl of
oclreolide on Iasma concenlralions of gIucose, insuIin, gIucagons, grovlh hormone, and
corlisoI in heaIlhy dogs and dogs vilh insuIinoma. Res Vel Sci 80:25-32.
35. SeIman I}, MoI }A, Rulleman GR, Ri|nberk A 1991 Irogeslins and grovlh hormone excess
in lhe dog. Acla IndocrinoI 125(suI 1):42-47.
36. MuIIigan T, }aen-VinuaIes A, GodschaIk M, Iranmanesh A, VeIdhuis }D 1999 Synlhelic
somaloslalin anaIog (oclreolide) suresses daylime grovlh hormone secrelion
equivaIenlIy in young and oIder men: reserved iluilary resonsiveness lo somaloslalin's
inhibilion in aging. } Am Geriar Soc 47:1422-1424.
37. Khava|a NM, Taher M, arham MI, Naser AA, Hadidy AM, Ahmad AT, Hamamy HA,
Yaghi NA, A|Iouni KM 2006 Iiluilary enIargemenl in alienls vilh hyolhyroidism.
Indocr Iracl 12:29-34.
38. Dam|anovic S, Ioovic V, Ielakov M, D|urovic M, Dieguez C, Casanueva II 1996 Iiluilary
enIargemenl due lo rimary hyolhyroidism: Grovlh hormone resonse lo GHRH,
GHRI-6 and GHRH Ius GHRI-6. } Iedialr Indocr Melab 9:549-553.
39. ei|erink N}, ui|leIs }}CWM, Okkens AC, Kooislra HS, DieIeman S} 2007 asaI and GnRH-
induced secrelion of ISH and LH in aneslrous versus ovarieclomized bilches.
TheriogeneIogy 67:1039-1045.
IunclionaI and morhoIogicaI changes in lhe adenohyohysis
_________________________________________________________________________________

143

40. Haon M, Molla A, Izquer M, onafede M, }ahn GA 2007 Hyolhyroidism roIongs
corus Iuleum funclion in lhe regnanl ral. Reroduclion 133:197-205.
41. Sharma Y, a|ai A, MillaI S, Kabra M, Menon IS 2006 Ovarian cysls in young girIs vilh
hyolhyroidism: foIIov-u and effecl of lrealmenl. } Iedialr IndocrinoI Melab19:895-900.
42. Mann DR, hal GK, Slah CD, IohI CR, IIanl TM 2006 Induclion of a hyolhyroid slale
during |uveniIe deveIomenl deIays uberlaI reaclivalion of lhe neuroendocrine syslem
governing Iuleinising hormone secrelion in lhe maIe rhesus monkey (Macaca muIalla). }
NeuroendocrinoI 18:662-671.
43. ZeIissen IM}, Croughs R}M, Koeschaar HII, de ruin TW, Hendriks M}, van 'l VerIaal
}W 1989 Irimary hyolhyroidism mimicking iluilary adenoma. Nelh } Med 35:95-99.
44. ZaIlsman I, Grimberg H, Luu-Meiri M, Lifschilz I, Oron Y 2000 Raid desensilizalion of
lhe TRH recelor and ersislenl desensilizalion of ils consliluliveIy aclive mulanl. ril }
IharmacoI 130:315-320.
45. IngeI S, Neumann S, Kaur N, Monga V, RahuI }, Norlhu }, Gershengorn MC 2006 Lov
affinily anaIogs of lhyrolroin-reIeasing hormone are suer-agonisls. } ioI Chem
281:13103-13109.
46. ViIIaIobos C, Nuez L, Garcia-Sancho } 2004 Anlerior iluilary lhyrlroes are
muIlifunclionaI ceIIs. Am } IhysioI IndocrinoI Melab 287:I1166-I1170.



Chaler 7
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144










Chaptcr 8

Adcnnhypnphysca! Iunctinn in dngs
with primary hypnthyrnidism
and nnn-thyrnida! i!!ncss








M.M. Diaz-Espincira, J.A. Mn!, A. Ri|nbcrk, H.5. Knnistra

Acccptcd Inr pub!icatinn in Jnurna! nI Vctcrinary Intcrna! Mcdicinc

_________________________________________________________________________

Dearlmenl of CIinicaI Sciences of Comanion AnimaIs,
IacuIly of Velerinary Medicine, Ulrechl Universily, Ulrechl, The NelherIands.

Chaler 8
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146

#

A recenl sludy of dogs vilh induced rimary hyolhyroidism (IH)
demonslraled lhal lhyroid hormone deficiency Ieads lo Ioss of lhyrolroin (TSH)
hyersecrelion, hyersomalolroism, hyoroIaclinemia, and iluilary
enIargemenl vilh Iarge vacuoIaled lhyroid deficiency ceIIs lhal doubIe-slained
for grovlh hormone (GH) and TSH, indicalive of lransdifferenlialion of
somalolroes lo lhyrosomalroes. Wilh lhe hyolhesis lhal simiIar funclionaI
changes in adenohyohyseaI funclion occur in dogs vilh sonlaneous IH as do
in dogs vilh induced IH, bul nol in dogs vilh non-lhyroidaI iIIness (NTI), lhe
adenohyohyseaI funclion vas invesligaled in 14 dogs vilh sonlaneous IH and
13 dogs vilh NTI, by combined inlravenous adminislralion of 4 hyohysiolroic
reIeasing hormones (4RH lesl), foIIoved by measuremenl of Iasma concenlralions
of adenocorlicolroic hormone (ACTH), GH, Iuleinizing hormone (LH), roIaclin
(IRL), and TSH. In lhe IH dogs lhis lesl vas reealed afler 4 and 12 veeks of
lhyroxine lrealmenl.
In 6 IH dogs, lhe basaI TSH concenlralion vas vilhin lhe reference range.
In lhe IH dogs, lhe TSH concenlralions did nol increase vilh lhe 4RH lesl.
Hovever, TSH concenlralions increased significanlIy in lhe NTI dogs. asaI and
slimuIaled GH and IRL concenlralions indicaled reversibIe hyersomalolroism
and hyerroIaclinemia in lhe IH dogs, bul nol in lhe NTI dogs. asaI and
slimuIaled LH and ACTH concenlralions did nol differ belveen grous.
Dogs vilh sonlaneous IH hyersecrele GH bul have IillIe or no TSH
hyersecrelion. DeveIomenl of hyerroIaclinemia (and ossibIe gaIaclorrhea) in
dogs vilh IH seems lo occur onIy in sexuaIIy inlacl bilches. In lhis grou of dogs
vilh NTI, basaI and slimuIaled Iasma adenohyohyseaI hormone
concenlralions are nol aIlered.
AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


147

Intrnductinn

In lhe 1960s and lhe earIy 1970s, lhe diagnosis of rimary hyolhyroidism
(IH) in dogs had lo reIy on (cumbersome) measuremenls of iodine (rolein-bound
iodine, II) in Iasma or lhe use of radioiodine. Wilh regard lo lhe Ialler
melhod, lhe abiIily of lhe lhyroid lo concenlrale iodine vas measured by
moniloring radioaclivily in lhe neck for lhree lo four days. The rocedure vas
reealed afler slimuIalion vilh bovine lhyroid-slimuIaling hormone (bTSH). This
aroach vas lime-consuming, exensive, and Iimiled lo research inslilulions.
Thyroid scanning vilh lhe shorl-Iived radionucIide lechnelium (
99m
TcO4
-
)
shorlened lhe rocedure considerabIy, bul sliII radialion recaulions vere
required. The advenl of radioimmunoassays for lhyroxine (T4) in lhe 1970s made
lhe diagnosis of hyolhyroidism easier. IH vas diagnosed vhen Iasma T4
concenlralion vas Iov and couId nol be raised sufficienlIy afler slimuIalion vilh
bTSH.
1,2,3

The inlroduclion of homoIogous immunoassays for measuring Iasma
concenlralions of TSH in lhe 1990s meanl lhal il vas ossibIe lo assess lhe canine
iluilary-lhyroid axis by measuring lhe hormone air T4 and TSH, vilh lhe
diagnosis of IH being eslabIished on lhe basis of a Iov Iasma T4 concenlralion
and a high Iasma TSH concenlralion. Hovever, in evaIualions vilh lhe TSH-
slimuIalion lesl as a goId slandard, il aeared lhal in u lo 33% of dogs vilh IH,
Iasma TSH concenlralions vere nol increased.
4,5,6
Due lo Iimilalions of lhe
endogenous canine TSH assay,
7
cIinicians have resorled lo using lhe lradilionaI
TSH slimuIalion lesl, using recombinanl human (rh) TSH inslead of bTSH.
8,9,10

Hovever, lhe goId slandard slalus of lhe TSH slimuIalion lesl has been
queslioned.
11
Techniques nol invoIving a biochemicaI assessmenl of lhe iluilary-
lhyroid axis, such as radionucIide imaging or ulake of
99m
TcO4
-
and high-
resoIulion uIlrasonograhy, seem lo be lhe mosl reIiabIe melhods for diagnosing
IH in dogs.
11,12

SeveraI assay modificalions have been suggesled lo imrove lhe Iov
diagnoslic vaIue of TSH measuremenls.
7
Hovever, lhere is nov evidence lhal il is
nol necessariIy lhe assay bul changes in iluilary funclion lhal are resonsibIe for
lhe Iov TSH concenlralions found in some dogs vilh IH. In a recenl sludy, 7
beagIe dogs vilh induced IH vere foIIoved for 3 years. Three of lhese dogs lhen
received I-lhyroxine for an addilionaI 1.5 years. Induclion of IH Ied lo high Iasma
TSH concenlralions for a fev monlhs, bul lhereafler concenlralions graduaIIy
decIined lo resuIls no Ionger significanlIy differenl from lhose of eulhyroid dogs. A
bIunled resonse lo slimuIalion of TSH reIease receded lhis decIine.
13
In
agreemenl vilh earIier observalions,
14
basaI Iasma grovlh hormone (GH)
concenlralions graduaIIy increased in associalion vilh lhe deveIomenl of a
Chaler 8
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148

aradoxicaI GH resonse lo adminislralion of lhyrolroin-reIeasing hormone
(TRH) in lhese hyolhyroid dogs. asaI Iasma roIaclin (IRL) concenlralions
decreased during hyolhyroidism, vhereas Iasma concenlralions of Iuleinizing
hormone (LH) and adrenocorlicolroic hormone (ACTH) vere essenliaIIy
unchanged. The funclionaI changes vere associaled vilh iluilary enIargemenl,
hisloIogicaIIy characlerized by lhyrolroe hyerIasia, Iarge vacuoIaled lhyroid
deficiency ceIIs, and doubIe-slaining of ceIIs indicalive of lransdifferenlialion of
somalolroes lo lhyrosomalolroes. Iiluilary enIargemenl and severaI of lhe
funclionaI changes vere reversibIe on lrealmenl vilh I-lhyroxine.
13
ResuIls of lhis
sludy cIarify lhe difficuIlies lhal may be encounlered vhen lrying lo diagnose IH
by measuring Iasma TSH concenlralion and confirm lhal canine IH is associaled
vilh hyersomalolroism. The abrulIy induced form of hyolhyroidism in
ovarieclomized beagIe dogs hovever may nol be reresenlalive of lhe graduaIIy
deveIoing sonlaneous disease in dogs of olher breeds, ages, and sex.
Here, ve reorl on lhe resuIls of adenohyohyseaI funclion lesls in 14
dogs vilh sonlaneous IH. ResuIls are comared vilh lhose of 13 dogs vilh Iov
Iasma T4

concenlralions, bul no lhyroid Iesions (i.e., dogs vilh non-lhyroidaI
iIIness |NTIj). In lhe IH dogs, funclionaI sludies vere conlinued during
subslilulion vilh I-lhyroxine.


Matcria! and Mcthnds


The 27 dogs sludied have been described in delaiI earIier.
11
riefIy, lhe 14
hyolhyroid dogs vere of various breeds and lheir ages ranged from 2 lo 10 years
(median, 5 years). Nine dogs vere maIe (3 caslraled) and 5 dogs vere femaIe (1
sayed). The median body veighl of lhe dogs vas 30 kg (range, 5-57 kg). IIeven
dogs veighed more lhan 25 kg. The diagnosis of IH vas based on Iov Iasma
lolaI T4 concenlralions (TT4, 2-4 nmoI/I |0.16-0.31 g/dIj), Iack of resonse lo TSH
slimuIalion, Iov lhyroid ulake of
99m
TcO4
-
, and exlensive Ioss of lhyroid lissue on
hisloalhoIogicaI examinalion of biosy samIes of lhe lhyroid gIand.
11
In 11 dogs,
generaIized infiIlralion of Iymhocyles and occasionaIIy Iasma ceIIs vas observed
in lhe biosy secimens. The remaining 3 dogs had comIele alrohy of lhyroid
lissue vilhoul infIammalory infiIlralion.
The 13 dogs vilh NTI vere of various breeds and lheir ages ranged from 3
lo 11 years (median, 6 years). Nine dogs vere maIe (3 caslraled) and 4 dogs vere
femaIe (2 sayed). The median body veighl vas 39 kg (range, 7-56 kg). TveIve
dogs veighed more lhan 25 kg. IIasma TT4 concenlralions ranged from 2-18 nmoI/I
(0.16-1.40 g/dI). IH vas excIuded by resuIls of lhyroid ulake of
99m
TcO4
-

AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


149

and hisloalhoIogic examinalion of lhyroid biosy samIes.
11
AII 13 dogs had
normaI lhyroid lissue vilh lhe usuaI varialion in foIIicIe size. The eilheIiaI ceIIs
vere cubicaI, vilh some resorlion vacuoIes in lhe ad|acenl coIIoid.

|unciicn icsis
In each dog, lhe rolocoI vas carried oul on 2 searale days vilh al Ieasl 1
veek (range, 1-3 veeks) belveen lesling. Al 8:00 a.m. on day 1, bIood samIes
vere coIIecled by cehaIic veniunclure for delerminalion of TT4 and TSH
concenlralions. ImmedialeIy lhereafler, quanlilalive ulake of
99m
TcO4
-
by lhe
lhyroid vas measured al 10, 20, 30, 45, 60, 90, and 120 minules afler in|eclion of lhe
radioharmaceulicaI.
11

On lhe second day, anlerior iluilary funclion vas invesligaled using
combined adminislralion of 4 hyohysiolroic reIeasing hormones: corlicolroin-
reIeasing hormone (CRH), grovlh hormone-reIeasing hormone (GHRH),
gonadolroin-reIeasing hormone (GnRH), and lhyrolroin-reIeasing hormone
(TRH) vilh measuremenl of ACTH, GH, LH, IRL, and TSH. This combined
anlerior iluilary funclion lesl (4RH lesl) vas erformed belveen 8:00 a.m. and
12:00 .m. afler an overnighl fasl, according lo melhods described reviousIy.
15,16

An inlravenous (iv) calheler vas Iaced in lhe cehaIic vein of each dog lo
faciIilale raid sequenliaI in|eclions. ImmedialeIy afler coIIeclion of lhe lime zero
bIood samIe from lhe |uguIar vein, lhe 4 hyolhaIamic reIeasing hormones vere
in|ecled iv, vilhin 30 seconds, al doses 1 g oCRH
a
er kg body veighl, 1 g
hGHRH
b
er kg body veighl, 10 g GnRH
c
er kg body veighl, and 10 g TRH
d

er kg body veighl. Iood samIes for delerminalion of Iasma concenlralions of
ACTH, GH, LH, IRL, and TSH vere coIIecled al 15, 0, 5, 10, 20, 30, and 45 min
from lhe |uguIar vein, immedialeIy Iaced in chiIIed IDTA-coaled or hearinized
(for TSH) lubes, and cenlrifuged al 4C for 10 minules al 3000 g. IIasma vas slored
al 25C unliI assayed.
Aflervards, a surgicaI lhyroid biosy for hisloalhoIogicaI examinalion of
lhyroid lissue vas laken.
11

Hcrncnc !cicrninaiicns
IIasma TT4 concenlralions vere delermined using a homoIogous soIid-
hase, chemiIuminescence enzyme immunoassay
e
in accordance vilh lhe
inslruclions of lhe manufaclurer and vaIidaled for use in lhe dog.
17
The inlra-assay
coefficienls of varialion (CVs) vere 13.8% and 8.2% al TT4 concenlralions of 8 and
25 nmoI/I, resecliveIy. The inler-assay CV vas 8.5% al a TT4 concenlralion of 21
nmoI/I. The Iovesl deleclabIe concenlralion of TT4 vas 2 nmoI/I.

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150

IIasma TSH concenlralions vere delermined vilh a homoIogous soIid-
hase, 2-sile chemiIuminescence enzyme immunomelric assay
f
in accordance vilh
lhe inslruclions of lhe manufaclurer and as described reviousIy.
17
The inlra-assay
CVs vere 5.0%, 4.0%, and 3.8% al TSH concenlralions of 0.20, 0.50 and 2.6 g/I,
resecliveIy. The inler-assay CVs vere 6.3% and 8.2% al TSH concenlralions of
0.16 and 2.8 g/I, resecliveIy. The Iovesl deleclabIe concenlralion of TSH vas 0.03
g/I. The uer Iimil of lhe reference range for lhe Iasma TSH concenlralion in
eulhyroid dogs in our Iaboralory is 0.6 g/I. TSH concenlralions beIov 0.03 g/I
are exressed as 0.03 g/I.
IIasma GH concenlralions vere delermined vilh a homoIogous
radioimmunoassay (RIA) as described reviousIy.
18
The inlra- and inler-assay CVs
vere 3.8% and 7.2%, resecliveIy. The sensilivily of lhe assay vas 0.3 g/I.
IIasma IRL concenlralions vere delermined vilh a reviousIy vaIidaled
heleroIogous RIA.
19
The inlra-assay and inler-assay CVs vere 3.5% and 11.5%,
resecliveIy. The Iover Iimil of quanlilalion vas 0.8 g/I, vhereas lhe maximum
Iimil of quanlilalion vas 200 g/I.
IIasma LH concenlralions vere delermined vilh a heleroIogous RIA
described earIier.
20
We used rabbil anliserum raised againsl ovine LH
g
, radio-
iodinaled bovine LH-7981 as reared for a bovine assay,
21
and canine iluilary
slandard LIR 1685-1
h
. The inlra-assay and inler-assay CVs for concenlralions
above 0.5 g/I vere 2.3% and 10.5%, resecliveIy. The Iimil of quanlilalion vas 0.3
g/I.
IIasma ACTH concenlralions vere delermined vilh a commerciaIIy
avaiIabIe 2-sile immunoradiomelric assay (IRMA) for use in humans
i
. The inlra-
and inler- assay CVs vere 3.2% and 7.8%, resecliveIy. The sensilivily vas 1 ng/I.

Trcaincni an! jc||cu-up
Afler IH vas diagnosed, aII 14 dogs vere lrealed vilh I-lhyroxine al an
iniliaI dosage of 10 g er kg body veighl lvice daiIy. Al foIIov-u examinalions
afler 4 and 12 veeks of subslilulion lheray, informalion on behavioraI and
hysicaI changes vas documenled. Al each foIIov-u examinalion, bIood samIes
vere coIIecled for measuremenl of Iasma concenlralions of TT4 and TSH and a
4RH slimuIalion lesl (see above) vas erformed. In lhe evenl of hysicaI or
biochemicaI evidence of undersubslilulion, lhe lhyroxine dose vas increased,
vhich occurred in 3 dogs, Ieading lo a range of dosages of 10-15 g/kg lvice daiIy.
The 4RH lesl vas erformed al 8:00 a.m., aroximaleIy 12 hours afler lhe Iasl
lhyroxine dose vas adminislered. ComIele foIIov-u dala vere avaiIabIe for 10
of lhe 14 dogs.


AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


151

AIready al lhe firsl foIIov-u examinalion, aII IH dogs had become more
aIerl, aclive, and resonsive. Wilh lime, lhere vas some decrease in body veighl
and comIele regrovlh of lhe hair coal.

Oaia ana|qsis
The area under lhe curve (AUC) above basaI hormone concenlralions vas
caIcuIaled using lhe lraezoidaI ruIe. The KoImogorov-Smirnov lesl vas used lo
lesl lhe normaIily of dala. ody veighls vere comared vilh Sludenl's i-lesl for
aired samIes. asaI hormone concenlralions and AUCs before and afler 4 and 12
veeks of subslilulion lheray vilh I-lhyroxine vere comared vilh lhe GeneraI
Linear ModeI MuIlivariale Tesl of SISS for Windovs (version 12.0.1, SISS Inc.,
Chicago, USA). Differences in basaI Iasma TSH, GH, IRL, and LH concenlralions,
as veII as differences in AUCs of TSH, GH, LH, and ACTH, belveen lhe
hyolhyroid dogs and lhe dogs vilh NTI vere comared vilh a Sludenl's i-lesl.
Differences in basaI Iasma ACTH concenlralions and AUCs of IRL belveen lhe
hyolhyroid dogs and dogs vilh NTI vere comared vilh lhe Mann-Whilney U-
lesl. Differences vere considered significanl al Is0.05. Dala are resenled as
means + SIM.


Ethics

The Ilhics Commillee of lhe IacuIly of Velerinary Medicine, Ulrechl
Universily aroved lhe sludies.

Rcsu!ts

Basa| ccnccniraiicns an! cjjccis cj 4|H siinu|aiicn
In lhe 14 IH dogs, lhe basaI Iasma TSH concenlralions ranged from 0.03
3.8 g/I vilh a mean (+ SIM) of 1.05 0.29 g/I. In 8 of lhese dogs, lhe Iasma TSH
concenlralion vas above lhe uer Iimil of lhe reference range, and 6 dogs had
Iasma TSH concenlralions vilhin lhe reference range (0.000.60 g/I). The Iasma
TSH concenlralions of lhe hyolhyroid dogs vere significanlIy (I0.016) higher
lhan lhose of lhe dogs vilh NTI (0.24 0.07 g/I). In lhe IH dogs, lhe 4RH lesls
caused no significanl change in Iasma TSH concenlralion, vhereas in lhe NTI
dogs a significanl increase occurred (Iig 1). In lhe NTI dogs, lhe mean AUC (above
baseIine) for TSH (14.85 2.27 g/I x 45 min) vas significanlIy higher (I0.001)
lhan lhal of lhe hyolhyroid dogs (4.35 1.74 g/I x 45 min).
The basaI Iasma GH concenlralions of lhe IH dogs (3.2 0.7 g/I) vere
significanlIy (I0.025) higher lhan lhose of lhe NTI dogs (1.4 0.3 g/I). In lhe IH
Chaler 8
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152

dogs, 4RH adminislralion increased lhe Iasma GH concenlralions significanlIy al
aII lime oinls, vhereas in lhe NTI dogs a significanl increase onIy occurred al 2
lime oinls earIy afler adminislralion of lhe reIeasing hormones (Iig 1). The mean
AUCs (above baseIine) of GH of lhe dogs vilh NTI (133 + 51 g/I x 45 min) and lhe
IH dogs (241 + 61 g/I x 45 min) vere nol significanlIy differenl (I0.19).
The mean basaI Iasma IRL concenlralion of lhe hyolhyroid dogs (3.9
1.3 g/I) vas higher bul nol significanlIy differenl (I0.13) from lhal of lhe dogs
vilh NTI (1.8 0.2 g/I). Afler removaI of lhe dala of lhe 2 bilches in meleslrus, lhe
IRL concenlralions of lhe dogs vilh rimary hyolhyroidism (2.5 0.2 g/I) vere
significanlIy (I0.03) higher lhan lhose of lhe dogs vilh NTI. In bolh grous, 4RH
adminislralion vas foIIoved by a non-significanl increase (I0.06 in IH dogs and
I0.10 in NTI dogs) in Iasma IRL concenlralions (Iig 1). The mean AUC (above
baseIine) in lhe NTI grou (273 97 g/I x 45 min) vas nol significanlIy differenl
(I0.22) from lhal in lhe hyolhyroid grou (1140 577 g/I x 45 min). The Iack of
a significanl resonse may have been due lo lhe Iarge inler-individuaI difference in
resonse (see Iig 1). This vas arlicuIarIy lrue for lhe IH dogs, 2 of vhich had
slrong IRL resonses lo 4RH slimuIalion. olh dogs vere inlacl femaIes lhal,
desile IH, sliII came inlo eslrus. Al lhe lime of lhe firsl 4RH lesl, 1 dog had |usl
been in eslrus, vhereas lhe olher |usl had slarled eslrus (Iig 2).
The mean basaI Iasma LH concenlralion of lhe hyolhyroid dogs (10.7
2.5 g/I) vas nol significanlIy differenl (I0.5) from lhal of lhe dogs vilh NTI (8.5
2.1 g/I). In bolh grous of dogs, Iasma LH concenlralions increased
significanlIy al aII lime oinls afler 4RH slimuIalion (Iig 1). The mean AUCs
(above baseIine) of lhe NTI dogs (588 98 g/I x 45 min) and lhe IH dogs (1308
496 g/I x 45 min) vere nol significanlIy differenl (I0.18).
The mean basaI Iasma ACTH concenlralion of lhe IH dogs (110 31 ng/I)
vas nol significanlIy differenl (I0.59) from lhal of lhe NTI dogs (79 22 ng/I). In
bolh grous, Iasma ACTH concenlralions increased significanlIy al aII lime oinls
afler 4RH slimuIalion (Iig 1). The mean AUCs (above baseIine) of lhe NTI dogs
(9089 1971 ng/I x 45 min) and lhe IH dogs (8410 1657 ng/I x 45 min) did nol
differ significanlIy (I0.79).
AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


153



Figurc 1. Mean ( SIM) Iasma concenlralions of TSH, GH, IRL, LH, and ACTH during lhe 4RH-
slimuIalion lesl in 14 dogs vilh rimary hyolhyroidism (vhile shere), and in 13 dogs vilh NTI (bIack
shere). Al lime oinl 0, 4RH slimuIalion vas erformed by inlravenous adminislralion of lhe 4
hyolhaIamic hormones TRH, GHRH, GnRH, and CRH. SignificanlIy higher lhan baseIine vaIues.
Chaler 8
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154






























Figurc 2. Logarilhmic Iols of Iasma IRL concenlralions measured during lhe 4RH-slimuIalion lesl in
2 femaIe dogs vilh IH. On admission (uer aneI) dog A () had slarled a nev eslrous cycIe 3 monlhs
earIier, vhiIe in dog () a nev eslrous cycIe |usl had slarled. Dog A had high basaI IRL
concenlralions vilh excessive resonsiveness lo slimuIalion. Dog had basaI IRL vaIues in lhe uer
range for heaIlhy aneslrus dogs (3.3 0.3 g/I),
35
and a resonse lhal |usl exceeded lhe range of eak
concenlralions of heaIlhy dogs (3.3 63.6 g/I).
23
Iour veeks Ialer (middIe aneI), vhiIe lhe dogs vere
on lhyroxine lheray, lhe silualion had reversed. In dog A bolh lhe basaI vaIues and lhe slimuIaled
vaIues had normaIized, and in dog basaI IRL concenlralions vere eIevaled vilh a cIear-cul resonse
lo slimuIalion aIbeil vilhin lhe reference range. Afler 12 veeks (Iover aneI) bolh lhe basaI and
slimuIaled IRL concenlralions of dog A vere Iov bul vilhin lhe reference range. In dog basaI IRL
vaIues had decreased and lhe resonse lo slimuIalion vas normaI.
AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


155

|jjccis cj |-inqrcxinc ircaincni in nqpcinqrci! !cgs
In 10 IH dogs, 4RH slimuIalion lesls vere erformed afler 4 and 12 veeks
of lheray vilh I-lhyroxine. The mean basaI Iasma TSH concenlralion decreased
significanlIy afler 4 and 12 veeks of lrealmenl vilh I-lhyroxine (I0.021 and
I0.05, resecliveIy) (Iig 3), vilh TSH concenlralions being beIov lhe uer Iimil
of lhe reference range (0.60 g/I) in 8 of 10 dogs afler 12 veeks. The 4RH
slimuIalion lesl did nol resuIl in a significanl increase in Iasma TSH concenlralion
in lhe lrealed dogs, and lhe mean AUC (above baseIine) of TSH did nol change
significanlIy comared vilh lhe re-lrealmenl resuIl (TabIe 1).
The mean basaI Iasma GH concenlralion decreased significanlIy afler 4
and 12 veeks of lrealmenl vilh I-lhyroxine (I0.034 and I0.024, resecliveIy) (Iig
3). Afler 4 veeks of lrealmenl vilh I-lhyroxine, 4RH slimuIalion resuIled in a
significanl increase (I0.02) in Iasma GH concenlralions al aII lime oinls,
vhereas afler 12 veeks a significanl increase onIy occurred 10 minules afler
slimuIalion. The mean AUC (above baseIine) of GH vas Iover (I0.032) afler 4
veeks of I-lhyroxine lrealmenl lhan before lrealmenl, vhereas afler 12 veeks lhe
mean AUC of GH did nol differ significanlIy from lhe mean AUC of GH before
lheray (TabIe 1).
The mean basaI Iasma IRL concenlralion did nol change significanlIy
afler 4 and 12 veeks of I-lhyroxine lrealmenl (Iig 3). Afler 4 veeks of lrealmenl,
lhe 4RH lesl resuIled in a significanl increase in Iasma IRL concenlralions al 5
and 10 minules afler slimuIalion (I0.02, and I0.03, resecliveIy), vhereas afler
12 veeks lhe increase in Iasma IRL concenlralions did nol reach slalislicaI
significance. In lhe 2 bilches lhal had recenlIy slarled a nev eslrous cycIe, lhe basaI
Iasma IRL concenlralions and lhe slimuIaled IRL concenlralions corresonded
lo lhe reseclive slages of lhe eslrous cycIes (Iig 2). The mean AUCs (above
baseIine) of IRL afler 4 and 12 veeks of lrealmenl vilh I-lhyroxine vere nol
significanlIy differenl from lhe re-lrealmenl resuIl (TabIe 1).
The mean basaI Iasma concenlralions of LH and ACTH did nol change
significanlIy during lhyroxine subslilulion (Iig 3). Afler bolh 4 and 12 veeks of
lrealmenl vilh I-lhyroxine, 4RH slimuIalion resuIled in a significanl increase in
Iasma concenlralions of LH and ACTH al aII lime oinls. The mean AUCs (above
baseIine) of LH, and ACTH afler 4 and 12 veeks of lrealmenl vilh I-lhyroxine
vere nol significanlIy differenl from lhe corresonding re-lrealmenl resuIls
(TabIe 1).
Chaler 8
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156






































Figurc 3. Mean ( SIM) basaI Iasma concenlralions of TSH, GH, IRL, LH, and ACTH of hyolhyroid
dogs, afler one and lhree monlhs of lheray vilh I-lhyroxine. SignificanlIy Iover lhan iniliaI vaIues.
AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


157



Hormone Basal After 4 weeks After 12 weeks
TSH (g/l) 1.2 0.39 0.6 0.29
*
0.44 0.21
*

GH (g/l) 3.4 0.9 1.2 0.4
*
0.9 0.3
*

PRL (g/l) 4.9 2.0 4.7 1.6 2.9 0.4
LH (g/l) 12.9 3.2 17.3 7.3 15.7 6.5
ACTH (ng/l) 94 32 112 29 95 20
AUC
TSH (g/l x 45 min) 5.1 2.3 6.3 2.9 5.8 2.0
GH (g/l x 45 min) 276 81 88 26
*
202 122
PRL (g/l x 45 min) 1501 790 180 79 197 94
LH (g/l x 45 min) 1712 658 1081 359 1381 508
ACTH (ng/l x 45 min) 9800 1967 13533 2241 11984 2230


Mean ( SIM) basaI hormone concenlralions and AUCs before and al 4 and 12
veeks of lrealmenl vilh I-lhyroxine in 10 dogs vilh rimary hyolhyroidism.

indicales
significanl difference comared vilh basaI vaIue.






Chaler 8
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158

Discussion

Use of lhe same 4RH slimuIalion lesl and hormone assays in lhe foIIov-u
sludy in dogs vilh induced IH
13
and in lhe resenl sludy in dogs vilh
sonlaneous disease rovides a good comarison. In lhe dogs vilh sonlaneous
IH, lhe lime from onsel of symloms of hyolhyroidism unliI resenlalion ranged
from 5 monlhs lo 4 years,
11
vilh a median of 1.5 years. Thus, for comarison vilh
lhe dogs vilh induced IH allenlion shouId be aid lo lhe changes lhal had
occurred 1-2 years afler induclion of hyolhyroidism, or even somevhal Ialer
because in severaI of lhe sonlaneous cases manifeslalions of lhe disease may have
Iagged behind deveIomenl of hyolhyroidism.
Afler an iniliaI increase by a faclor 15 in lhe dogs vilh induced IH and 1-2
years afler lhe induclion of hyolhyroidism, basaI Iasma TSH concenlralions had
decreased lo aroximaleIy lhe same concenlralion as measured in lhe dogs vilh
sonlaneous IH in lhe resenl sludy. olh in dogs vilh induced and sonlaneous
IH, lhere vas no significanl effecl of 4RH slimuIalion on Iasma TSH
concenlralions. In conlrasl, in lhe NTI dogs lhe Iover basaI Iasma TSH
concenlralions increased in lhe 4RH lesl in a manner simiIar lo lhal reorled
reviousIy for heaIlhy dogs.
22
Whereas in bolh grous of IH dogs, basaI Iasma
TSH concenlralions decreased addilionaIIy vilh lhyroxine lrealmenl, lhe 3 monlhs
of subslilulion lheray did nol reslore TSH resonsiveness in lhe 4RH lesl.
As for TSH, lhere aIso vere no nolabIe differences in GH belveen lhe
lvo IH grous. asaI Iasma GH concenlralions vere increased in lhe IH dogs
and increased addilionaIIy afler 4RH adminislralion. The Iover Iasma GH
concenlralions in lhe NTI dogs aIso increased in lhe 4RH lesl, aIlhough Iess lhan in
lhe IH dogs. The higher resonse of lhe IH dogs may have been caused by a
aradoxicaI resonse lo TRH,
14
as veII as a resonse lo GHRH. Whereas in lhe NTI
dogs, as in heaIlhy dogs, lhere vas mosl IikeIy onIy a resonse lo GHRH.
14,23
As
observed in lhe dogs vilh induced IH, in lhe dogs vilh sonlaneous IH Iasma
GH concenlralions decreased during lhyroxine lrealmenl.
Wilh regard lo IRL, lhe silualion vas quile differenl in lhe dogs vilh
induced IH as comared lo lhe dogs vilh sonlaneous IH. Afler 1-2 years, lhe
dogs vilh induced IH had significanlIy Iover Iasma IRL concenlralions lhan
iniliaIIy and aIso lhe IRL resonses lo 4RH slimuIalion had decreased
considerabIy. In conlrasl, in lhe dogs vilh sonlaneous IH lhe basaI IRL
concenlralions vere nol Iov and in generaI lhere vas a good resonse in lhe 4RH
lesl, aIbeil vilh vide varialion. The exIanalion for lhis difference may be lhal lhe
dogs vilh induced IH vere sayed femaIes, vhereas some of lhe dogs vilh
sonlaneous IH vere sexuaIIy inlacl. In arlicuIar, 2 dogs vilh sonlaneous IH

AdenohyohyseaI funclion in dogs vilh IH and NTI
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159

lhal recenlIy slarled a nev eslrous cycIe had or deveIoed high basaI IRL
concenlralions vilh slrong resonsiveness lo 4RH adminislralion.
The occurrence of hyerroIaclinemia (and gaIaclorrhea) has been
reorled earIier in a femaIe dog vilh rimary hyolhyroidism.
24
In humans,
aroximaleIy 8% of alienls vilh IH have increased Iasma IRL concenlralions
vilh gaIaclorrhea, vhich occurs eseciaIIy in regnanl or Iaclaling vomen.
25

Disinhibiled hyolhaIamic TRH slimuIalion and olher mechanisms, such as a Iack
of direcl effecls of lhyroxine on hyolhaIamic doaminergic mechanisms and
Iaclolroe doamine recelor funclion, have been suggesled lo cause lhe increase
in Iasma IRL concenlralion.
26
Ixogenous TRH is a olenl slimuIus of roIaclin
reIease,
27
bul accumuIaled dala for mice indicale lhal endogenous TRH in lhe
hyolhaIamus is nol invoIved subslanliaIIy in lhe mainlenance of IRL secrelion
under nonsuckIing condilions.
28
The recenlIy idenlified roIaclin-reIeasing elide
(IrRI) may have a hysioIogicaI roIe in roIaclin reIease.
29

In accordance vilh lhe occurrence of hyerroIaclinemia (and
gaIaclorrhea) in humans vilh IH during regnancy and Iaclalion, in lhe resenl
sludy hyerroIaclinemia occurred rimariIy in lhe IuleaI hase of lhe eslrous
cycIe of lhe IH dogs. There is evidence lhal lhe reguIalion of IRL secrelion is
deendenl on sleroid concenlralions. In femaIe rhesus monkeys, lhe reguIalion of
IRL reIease during lhe IuleaI hase is differenl from lhal observed in lhe foIIicuIar
and menslruaI hases. The reIease of IrRIs and IRL may be enhanced in lhe
resence of ovarian feedback.
30
In dogs, rogeslerone in arlicuIar has a
moduIaling roIe in roIaclin secrelion.
31

The imorlanl roIe of rogeslerone comared vilh olher gonadaI sleroids
is consislenl vilh lhe Iack of lhe occurrence of hyerroIaclinemia or gaIaclorrhea
in maIe dogs vilh IH. None of lhe inlacl maIe dogs vilh sonlaneous IH in lhe
resenl sludy deveIoed hyerroIaclinemia. In men, gaIaclorrhea and
gynecomaslia are regarded as very rare manifeslalions of IH.
32
In a Iarge series of
humans vilh IH (femaIe lo maIe ralio 13:1), hyerroIaclinemia occurred in 84 oul
of 1003 alienls (8%). Iive of lhese alienls vere men. In lhe men vilh IH and
hyerroIaclinemia, Iasma IRL concenlralions vere significanlIy Iover lhan in
vomen vilh IH and hyerroIaclinemia.
25

There is increasing evidence lhal in dogs il is nol an increase in lhe Iasma
rogeslerone concenlralion lhal slimuIales IRL reIease bul ralher a decrease in ils
concenlralion. In non-regnanl bilches, lhe decrease in lhe second arl of lhe IuleaI
hase is associaled vilh an increase in bolh basaI Iasma IRL concenlralion and
AUC for IRL.
31
AIso in seudoregnanl bilches, decreasing Iasma rogeslerone
concenlralions are associaled vilh increasing IRL reIease.
33
Moreover,
adminislralion of a rogeslerone recelor anlagonisl lo regnanl bilches causes
Iasma IRL concenlralions lo increase sharIy.
34
SimiIarIy, in lhe 2 hyolhyroid
Chaler 8
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160

bilches lhal recenlIy slarled a nev eslrous cycIe, lhe highesl basaI IRL
concenlralions and resonses lo slimuIalion occurred Iale in lhe IuleaI hase.
In dogs vilh induced IH,
13
and in lhe resenl dogs vilh sonlaneous IH
and NTI, basaI and slimuIaled Iasma concenlralions of LH and ACTH vere in
aroximaleIy lhe same range. In bolh lhe dogs vilh induced IH and lhe dogs
vilh sonlaneous IH, lhese variabIes vere nol affecled by lhyroxine subslilulion.
In concIusion, dogs vilh sonlaneous IH hyersecrele GH bul have IillIe
or no TSH hyersecrelion. The deveIomenl of hyerroIaclinemia (and ossibIy
gaIaclorrhea) in dogs vilh IH seems lo occur onIy in sexuaIIy inlacl bilches. In lhis
grou of dogs vilh NTI, basaI and slimuIaled Iasma adenohyohyseaI hormone
concenlralions are nol aIlered.

Acknnw!cdgmcnts

The aulhors gralefuIIy acknovIedge lhe heI of Mrs. M.I. van WoIferen
and lhe Iaboralory slaff. The heI of Dr. }. van den roek in lhe slalislicaI anaIysis
is highIy arecialed. The aulhors are very gralefuI for lhe skiIIed and enlhusiasl
assislance of Mrs. Y.W.I.A. IoIIak and Mr. H.G.H. van IngeIen.

AdenohyohyseaI funclion in dogs vilh IH and NTI
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161

References

1. uIIock L 1970 Irolein bound iodine delerminalion as a diagnoslic aid for canine
hyolhyroidism. } Am Vel Med Ass 156:892-899.
2. Ri|nberk A 1974 Hyolhyroidism. In: Currenl Velerinary Theray V. Kirk RW (Id.).
IhiIadeIhia, IA: W Saunders Comany. :791-797.
3. eIshav I, Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Am Anim Hos Ass 15:17-23.
4. Ielerson MI, MeIian C, NichiIs R 1997 Measuremenls of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroin concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Ass 211:1396-1402.
5. Dixon RM, Mooney CT 2001 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40:72-78.
6. orelli IS, Reusch CI 2004 Indogenous TSH in lhe diagnosis of hyolhyroidism in dogs.
Schveiz Arch TierheiIkd 146:183-188.
7. Ierguson DC 2007 Tesling for hyolhyroidism in dogs. Vel CIin Norlh Am SmaII Anim Iracl
37:647-669.
8. Sauve I, Iaradis M 2000 Use of recombinanl human lhyroid-slimuIaling hormone for
lhyrolroin slimuIalion lesl in eulhyroid dogs. Can Vel } 41:215-219.
9. orelli IS, Sieber-RucksluhI NS, Iavrol C, Lulz H, Hofmann-Lehmann R, Reusch CI 2006
IvaIualion of recombinanl human lhyroid-slimuIaling hormone lo lesl lhyroid funclion in
dogs susecled of having hyolhyroidism. Am } Vel Res 67:2012-2016.
10. De Roover K, Duchaleau L, CarmichaeI N, van Geffen C, Daminel S 2006 Iffecl of slorage of
reconsliluled recombinanl human lhyroid-slimuIaling hormone (rhTSH) on lhyroid-
slimuIaling hormone (TSH) resonse lesling in eulhyroid dogs. } Vel Inlern Med 20:812-817.
11. Diaz Isieira MM, MoI }A, Ieelers MI, IoIIak YWIA, Iversen L, van Di|k }I, Ri|nberk A,
Kooislra HS 2007 Assessmenl of lhyroid funclion in dogs vilh Iov Iasma lhyroxine
concenlralion. } Vel Inlern Med 21:25-32.
12. Reese S, reyer U, Deeg C, Krafl W, Kasers 2005 Thyroid sonograhy as an effeclive looI
lo discriminale belveen eulhyroid sick and hyolhyroid dogs. } Vel Inlern Med 19:491-498.
13. Diaz Isieira MM, MoI }A, van den Ingh TSGAM, van der VIugl-Mei|er RH, Ri|nberk A,
Kooislra HS 2008 IunclionaI and morhoIogicaI changes in lhe adenohyohysis of dogs
vilh induced rimary hyolhyroidism, Ioss of TSH hyersecrelion, hyersomololroism,
hyoroIaclinemia, and iluilary enIargemenl vilh lransdifferenlialion. Domesl Anim
IndocrinoI 35:98-111.
14. Diaz Isieira MM, GaIac S, MoI }A, Ri|nberk A, Kooislra HS 2008 Thyrolroin-reIeasing
hormone-induced grovlh hormone secrelion in dogs vilh rimary hyolhyroidism. Domesl
Anim IndocrinoI 34:176-181.


Chaler 8
_____________________________________________________________________________

162

15. Mei| I, MoI }A, HazevinkeI HAW, evers MM, Ri|nberk A 1996 Assessmenl of a combined
anlerior iluilary funclion lesl in beagIe dogs: raid sequenliaI inlravenous adminislralion of
four hyolhaIamic reIeasing hormones. Domesl Anim IndocrinoI 13:161-170.
16. Mei| I, MoI }A, Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13:465-468.
17. runer }, Scoll-Moncrieff CR, WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on serum
lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am Vel
Med Assoc 212:1572-1575.
18. Iigenmann }I, Iigenmann RY 1981 Radioimmunoassay of canine grovlh hormone. Acla
IndocrinoI,98:514-520.
19. Okkens AC, evers MM, DieIeman S}, WiIIemse AH 1985 Shorlening of lhe inleroeslrous
inlervaI and lhe Iifesan of lhe corus Iuleum of lhe cycIic dog by bromocriline lrealmenl.
Vel Quarl 7:173-176.
20. Nell TM, Akbar AM, Ihemisler RD, HoIsl IA, Reicherl }r LI, Nisvender GD 1975 LeveIs of
Iuleinizing hormone, eslradioI and rogeslerone in serum during lhe eslrous cycIe and
regnancy in lhe beagIe bilch. Iroc Soc Ix ioI Med 148:134-139.
21. DieIeman S}, evers MM 1987 Iffecls of monocIonaI anlibody againsl IMSG adminislered
shorlIy afler lhe reovuIalory LH surge on lime and number of ovuIalions in IMSG/IG-
lrealed covs. } Rerod IerliI 81:533-542.
22. Mei| I, MoI }A, Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13:465-468.
23. Mei| I, MoI }A, HazevinkeI HAW, evers MM, Ri|nberk A 1996 Assessmenl of a combined
anlerior iluilary funclion lesl in beagIe dogs: Raid sequenliaI inlravenous adminislralion of
four hyolhaIamic reIeasing hormones. Domesl Anim IndocrinoI 13:161-170.
24. Corlese L, OIiva G, Verslegen }, CiarameIIa I, Iersechino A 1997 HyerroIaclinaemia and
gaIaclorrhoea associaled vilh rimary hyolhyroidism in a bilch. } SmaII Anim Iracl 38:572-
575.
25. Raber W, GessI A, Novolny I, Vierhaer H 2003 HyerroIaclinaemia in hyolhyroidism:
cIinicaI significance and imacl of TSH normaIizalion. CIin IndocrinoI 58:185-191.
26. Conlreras I, Generini G, MicheIsen H, Iumarino H, Camino C 1981 HyerroIaclinemia
and gaIaclorhea: sonlaneous versus ialrogenic hyolhyroidism. } CIin IndocrinoI Melab
53:1036-1039.
27. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IMM,
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114:275-282.
28. Yamada M, Saloh T, Mori M 2003 Mice Iacking lhe lhyrolroin-reIeasing hormone gene:
Whal do lhey leII us` Thyroid 13:1111-1121.
AdenohyohyseaI funclion in dogs vilh IH and NTI
_____________________________________________________________________________


163

29. Sun , Iu|ivara K, Adachi S, Inoue K 2005 IhysioIogicaI roIes of roIaclin-reIeasing elide.
ReguI Iel 15:27-33.
30. }ahan S, }aIaIi S, Shami SA 2007 Neuroendocrine reguIalion of roIaclin secrelion in aduIl
femaIe rhesus monkeys during differenl hases of lhe menslruaI cycIe: roIe of neuroexcilalory
amino acid (NMA). Am } IrimaloI 69:395-406.
31. Kooislra HS, Okkens AC 2002 Secrelion of grovlh hormone and roIaclin during rogression
of lhe IuleaI hase in heaIlhy dogs: a reviev. MoI CeII IndocrinoI 197:167-172.
32. Arnaoul MA, Garlhvaile TL, Krubsack A}, Hagen TC 1987 GaIaclorrhea, gynecomaslia, and
hyolhyroidism in a man. Ann Inlern Med 106:779-780.
33. Okkens AC, DieIeman S}, Kooislra HS, evers MM 1997 IIasma concenlralions of roIaclin in
overlIy seudoregnanl Afghan hounds and lhe effecl of melergoIine. } Rerod IerliI SuI
51:295-301.
34. GaIac S, Kooislra HS, ulinar }, evers MM, DieIeman S}, Voorhoul G, Okkens AC 2000
Terminalion of mid-geslalion regnancy in bilches vilh agIerislone, a rogeslerone recelor
anlagonisl. TheriogenoIogy 53:941-950.
35. ei|erink N}, DieIeman S}, Kooislra HS, Okkens AC 2003 Lov doses of bromocriline shorlen
lhe inleroeslrous inlervaI in lhe bilch vilhoul Iovering Iasma roIaclin concenlralion.
TheriogenoIogy 60:1379-1386.



a
IeninsuIa Laboralories Inc., eImonl, CA, USA
b
IeninsuIa Laboralories Inc.
c
IerlagyI, Inlervel, oxmeer, The NelherIands
d
TRH Hoffmann-La Roche, aseI, SvilzerIand
e
ImmuIile canine TolaI T4, Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, CA, USA
f
ImmuIile canine TSH, Diagnoslic Iroducls Cororalion (DIC), Los AngeIes, CA, USA
g
CSU-204, kindIy suIied by G.D. Nisvender, CoIorado Slale Universily, CO, USA
h
a gifl of Dr L.I. Reicherl, AIbany MedicaI CoIIege, NY, USA
i
NichoIs Inslilule, Wi|chen, NL














Chaler 8
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164




Chaptcr 9

5ummarizing discussinn and cnnc!usinns
Chaler 9
_________________________________________________________________________________

166
Summarizing discussion and concIusions
_________________________________________________________________________________

167

Hyolhyroidism may be erroneousIy diagnosed in dogs because many non-
lhyroidaI faclors (e.g., drug lheray, concurrenl disease) may resuIl in a Iover
Iasma lolaI lhyroxine (TT4) concenlralion. Il vas hoed lhal lhe inlroduclion in
lhe 1990s of homoIogous immunoassays for measuring canine Iasma lhyroid-
slimuIaling hormone (TSH) concenlralions vouId imrove diagnoslic accuracy,
vilh lhe diagnosis of rimary hyolhyroidism being eslabIished on lhe basis of a
Iov Iasma TT4 concenlralion and a high Iasma TSH concenlralion. Hovever,
vhen lhe TSH-slimuIalion lesl vas used as a goId slandard, il aeared lhal
Iasma TSH concenlralions vere nol eIevaled in as many as a lhird of dogs vilh
rimary hyolhyroidism. The main aim of lhe sludies described in lhis lhesis vas
lo lesl lhe hyolhesis lhal rimary hyolhyroidism Ieads lo adenohyohyseaI
aIleralions lhal may exIain lhe occurrence of Iov Iasma TSH concenlralions. In
addilion, il vas hoed lhal lhe sludies vouId generale beller vays lo diagnose
rimary hyolhyroidism in dogs.
As menlioned above, Iasma TT4 concenlralions may be Iov in dogs vilh
non-lhyroidaI iIIness (NTI), and lherefore differenlialion belveen canine
hyolhyroidism and non-lhyroidaI iIIness may be difficuIl. Chaptcr 3 reorls on
lhe resuIls of a diagnoslic sludy invoIving 30 dogs vilh a Iov Iasma TT4
concenlralion. In lhese dogs lhe finaI diagnosis of rimary hyolhyroidism or non-
lhyroidaI iIIness vas based uon hisloalhoIogicaI examinalion of a lhyroid
biosy. AII dogs vilh rimary hyolhyroidism had very Iov Iasma TT4
concenlralions. This is consislenl vilh revious sludies reorling lhal
measuremenl of lhe Iasma TT4 concenlralion is a sensilive lesl for diagnosing
hyolhyroidism.
1-5
In lhe dogs vilh non-lhyroidaI iIIness, lhe Iasma TT4
concenlralions vere significanlIy higher lhan lhose in lhe dogs vilh rimary
hyolhyroidism, bul some of lhe dogs vilh non-lhyroidaI iIIness aIso had very Iov
Iasma TT4 concenlralions. This confirms lhal vhiIe lhe sensilivily of TT4
measuremenls in diagnosing hyolhyroidism is high, lhe secificily is Iov.
5,6

Serum free T4 (fT4) concenlralions in lhe dogs vilh rimary
hyolhyroidism vere significanlIy Iover lhan lhose in lhe dogs vilh non-lhyroidaI
iIIness. AII hyolhyroid dogs bul one had serum fT4 concenlralions beIov lhe
deleclion Iimil of lhe assay. On lhe olher hand, onIy one dog vilh non-lhyroidaI
iIIness had a serum fT4 concenlralion beIov lhe deleclion Iimil of lhe assay. The
sIighlIy beller secificily of fT4 comared vilh TT4 couId be considered a reason lo
incIude lhis lesl in rouline diagnoslics for canine hyolhyroidism,
4,7
afler
eslabIishmenl of a reference range in an aroriale number of dogs.
The TT4 incremenl afler TSH slimuIalion vas significanlIy higher in lhe
dogs vilh non-lhyroidaI iIIness lhan in lhe dogs vilh rimary hyolhyroidism. In
some of lhe dogs vilh non-lhyroidaI iIIness lhere vas onIy a minor increase in
Iasma TT4 concenlralions. If earIier eslabIished cul-off vaIues for hyolhyroidism
Chaler 9
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168

(osl-slimuIalion T4 >25 nmoI/I and/or al Ieasl 1.5 limes lhe baseIine concenlralion)
4

vouId have been aIied, some dogs vilh non-lhyroidaI iIIness vouId have been
miscIassified as having rimary hyolhyroidism. Il is IikeIy lhal lhis
miscIassificalion has occurred in revious sludies of canine hyolhyroidism in
vhich lhe TSH-slimuIalion lesl vas used as a goId slandard in order lo assess
olher lesls. NeverlheIess, lhe TSH-slimuIalion lesl may sliII be regarded as lhe Ieasl
inconcIusive biochemicaI lhyroid funclion lesl for differenlialing belveen rimary
hyolhyroidism and non-lhyroidaI iIIness in dogs.
A reIaliveIy high number of dogs vilh rimary hyolhyroidism (6 of 14)
had a Iasma TSH concenlralion vilhin lhe reference range for eulhyroid dogs.
This is in agreemenl vilh revious observalions (vilh lhe TSH-slimuIalion lesl as
lhe goId slandard) lhal Iasma TSH concenlralions are nol eIevaled in as many as a
lhird of dogs vilh rimary hyolhyroidism.
3,4,5,8,9,10
asaI Iasma TSH
concenlralions in lhe dogs vilh non-lhyroidaI iIIness vere vilhin lhe reference
range, vilh lhe excelion of one dog lhal had an eIevaled Iasma TSH
concenlralion. The occurrence of Iasma TSH concenlralions exceeding lhe
reference range has been reorled before in eulhyroid dogs.
3,4,5,10

Measuremenl of lhe TSH resonse afler lhyrolroin-reIeasing hormone
(TRH) adminislralion offers a raid, safe, and effeclive melhod of invesligaling lhe
reserve caacily of canine iluilary lhyrolroic ceII funclion.
11,12
TheorelicaIIy, lhe
TRH-slimuIalion lesl can aIso be used in lhe diagnosis of rimary hyolhyroidism,
because exogenous TRH may acl as an amIifier lo exaggerale an abnormaIily in
TSH secrelion.
13
Hovever, in lhe dogs vilh rimary hyolhyroidism lhere vas nol
an exaggeraled TSH resonse lo slimuIalion vilh TRH. On lhe conlrary, in Iine
vilh revious sludies,
3,14
lhe increase aeared lo be significanlIy smaIIer lhan lhal
in lhe dogs vilh non-lhyroidaI iIIness. AIso lhe resuIls of lhe sludy reorled in
Chaptcr 6 shoved lhal TRH adminislralion causes Iasma TSH concenlralions lo
rise significanlIy in heaIlhy conlroI dogs, bul nol in hyolhyroid dogs. These
observalions indicale lhal iluilary resonsiveness lo TRH is considerabIy
decreased in dogs vilh rimary hyolhyroidism. In conlrasl, lhe TSH resonse lo
TRH is aIvays increased in humans vilh rimary hyolhyroidism.
15
AIso human
alienls vilh Iong-slanding rimary hyolhyroidism and lhyrolroe hyerIasia
resond in an exaggeraled fashion lo TRH slimuIalion.
16
Thus lhe occurrence of
bIunled or absenl TSH resonses in lhe TRH lesl is secific lo hyolhyroid dogs,
and so far unexIained.
Al 30 minules afler TRH adminislralion, lhe Iasma TSH concenlralions
did nol differ significanlIy belveen hyolhyroid dogs and dogs vilh non-lhyroidaI
iIIness. This recIudes lhe use of lhe TRH-slimuIalion lesl vilh measuremenl of
Iasma TSH concenlralions lo dislinguish belveen rimary hyolhyroidism and
lhe non-lhyroidaI iIIness, as found earIier.
13,14

Summarizing discussion and concIusions
_________________________________________________________________________________

169

CaIcuIalion of lhe ercenl ulake of
99m
TcO4
-
inlo lhe lhyroid has roven lo
be very usefuI in lhe assessmenl of lhyroid funclion in humans and cals.
17,18
This
vas aIso lhe case for lhe resenl dogs vilh hyolhyroidism. In lhe erlechnelale
ulake sludies, lhere vas no overIa in lhyroid
99m
TcO4
-
ulake al 45-120 minules
afler in|eclion of lhe radioharmaceulicaI belveen lhe dogs vilh rimary
hyolhyroidism and lhe dogs vilh non-lhyroidaI iIIness. In lhe dogs vilh non-
lhyroidaI iIIness,
99m
TcO4
-
ulake vas comarabIe lo lhal reorled in heaIlhy beagIe
dogs.
19

The resuIls of lhe sludy reorled in Chaptcr 3 confirm earIier observalions
lhal in dogs accurale biochemicaI diagnosis of rimary hyolhyroidism oses
secific robIems and demonslrale lhal lhe circuIaling concenlralions of TT4, fT4,
and TSH overIa in rimary hyolhyroidism and non-lhyroidaI iIIness. ResuIls of
lhe TSH-slimuIalion lesl did nol meel earIier eslabIished diagnoslic crileria and/or
overIaed in lhe lvo grous of dogs, indicaling lhal in revious sludies, in vhich
lhe TSH-slimuIalion lesl vas used as lhe goId slandard for lhe diagnosis of
hyolhyroidism, dogs may have been miscIassified. Quanlilalive measuremenl of
lhe lhyroid ulake of
99m
TcO4
-
had lhe highesl discriminalory over vilh regard lo
lhe differenlialion belveen rimary hyolhyroidism and non-lhyroidaI iIIness.
To delermine vhelher Iasma TSH concenlralions vilhin lhe reference
range in dogs vilh rimary hyolhyroidism can be exIained by fIuclualions in
lhe iluilary reIease of TSH, lhe Iasma rofiIes of TSH vere anaIyzed in 7 beagIe
bilches by coIIecling bIood samIes every 10 minules for 6 hours, bolh before and
afler lhe induclion of rimary hyolhyroidism (Chaptcr 4). The resuIls
demonslrale uIsaliIe secrelion of TSH in dogs during hyolhyroidism and onIy
smaII fIuclualions in Iasma TSH concenlralion during eulhyroidism. The
fIuclualions around lhe TSH baseIine in lhe dogs vilh induced rimary
hyolhyroidism may in arl exIain vhy in some dogs vilh sonlaneous rimary
hyolhyroidism Iasma TSH concenlralions are vilhin lhe reference range for
eulhyroid dogs. The mean Iasma TSH concenlralion in dogs vilh sonlaneous
rimary hyolhyroidism has been reorled lo exceed onIy marginaIIy lhe uer
Iimil of lhe reference range for TSH in eulhyroid dogs.
4,10
ConsequenlIy,
fIuclualions around lhe TSH baseIine al lhis IeveI may occasionaIIy resuIl in
Iasma TSH concenlralions vilhin lhe reference range for eulhyroid dogs.
The cIinicaI manifeslalions of rimary hyolhyroidism in dogs incIude
veighl gain, Ielhargy, and dermaloIogicaI changes such as lruncaI aIoecia and
skin lhickening due lo mucooIysaccharide accumuIalion.
20,21
To lesl lhe ossibIe
invoIvemenl of aIlered secrelion of grovlh hormone (GH) in lhe fealures of dogs
vilh rimary hyolhyroidism, 6-hour uIsaliIe allerns of GH secrelion vere
invesligaled bolh before and afler induclion of hyolhyroidism in seven beagIe
dogs (Chaptcr 5). In conlrasl lo severaI olher mammaIian secies, in dogs rimary
Chaler 9
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170

hyolhyroidism aeared lo be associaled vilh eIevaled GH secrelion. This
eIevaled GH secrelion has endocrine reIevance, as iIIuslraled by eIevaled Iasma
insuIin-Iike grovlh faclor-I (IGI-I) concenlralions and some hysicaI changes
mimicking acromegaIy.
In eulhyroid humans, rals, and dogs, adminislralion of TRH does nol
resuIl in areciabIe changes in Iasma GH IeveIs.
22-26
In conlrasl, in humans and
rals vilh rimary hyolhyroidism, TRH adminislralion resuIls in significanlIy
increased Iasma GH IeveIs.
22-25
To invesligale vhelher adminislralion of TRH aIso
resuIls in a aradoxicaI rise in Iasma GH concenlralion in dogs vilh rimary
hyolhyroidism, lhe effecl of inlravenous adminislralion of TRH on GH reIease
vas sludied in len dogs vilh rimary hyolhyroidism and six heaIlhy conlroI dogs
(Chaptcr 6). In lhe dogs vilh rimary hyolhyroidism, lhe Iasma GH
concenlralion increased significanlIy afler in|eclion of TRH. As reorled earIier by
Rulleman
26
in lhe heaIlhy conlroI dogs lhe Iasma GH concenlralion did nol
increase significanlIy afler TRH adminislralion.
Massive lhyrolroe hyerIasia has been found in humans vilh rolracled
rimary hyolhyroidism. DoubIe immunoslaining, immunoeIeclron microscoy,
and in silu hybridizalion reveaIed lhe resence of bihormonaI ceIIs roducing bolh
GH and TSH.
27
Thus, in addilion lo lhe muIliIicalion of lhyrolroes,
lransdifferenlialion of somalolroes inlo lhyrolroes seems lo conlribule lo lhe
increase in TSH-roducing ceIIs. To invesligale vhelher simiIar morhoIogicaI
changes in lhe iluilary gIand aIso occur in dogs vilh rimary hyolhyroidism,
seven beagIe dogs vere foIIoved u for 3 years afler lhe induclion of rimary
hyolhyroidism (Chaptcr 7). Three of lhese dogs vere foIIoved u for anolher 1.5
years vhiIe receiving I-lhyroxine. AdenohyohyseaI funclion vas sludied al 2-
monlh inlervaIs vilh lhe combined inlravenous in|eclion of four hyolhaIamic
reIeasing hormones (CRH, GHRH, GnRH, and TRH) and measuremenl of lhe
Iasma concenlralions of lhe iluilary hormones ACTH, GH, LH, IRL, and TSH.
In addilion, every 6 monlhs lhe iluilary gIand vas visuaIized by comuled
lomograhy. HislomorhoIogicaI and immunohislochemicaI sludies of lhe
iluilary gIands vere erformed in dogs vilh unlrealed hyolhyroidism and in
hyolhyroid dogs on subslilulion lheray vilh I-lhyroxine.
The induclion of rimary hyolhyroidism caused lhe Iasma TSH
concenlralions lo rise by a faclor of aboul 15. These Iasma TSH concenlralions
remained al lhis high IeveI for onIy a fev monlhs and lhen graduaIIy decIined,
aIlhough remaining significanlIy higher lhan lhe iniliaI eulhyroid vaIues for mosl
of lhe observalion eriod. Hovever, al lhe end of lhis arl of lhe sludy (ca. 3 years
afler induclion of rimary hyolhyroidism), lhe Iasma TSH concenlralions vere
nol significanlIy differenl from lhe iniliaI vaIues. The TSH resonse lo
surailuilary slimuIalion decreased even fasler. These observalions can exIain
Summarizing discussion and concIusions
_________________________________________________________________________________

171

lhe oflen noneIevaled basaI Iasma TSH concenlralions measured in dogs vilh
sonlaneous rimary hyolhyroidism.
asaI Iasma GH concenlralions increased graduaIIy during lhe firsl year
and remained al high IeveIs. Thus in conlrasl lo lhe basaI TSH concenlralions, lhe
eIevaled basaI GH concenlralions did nol decIine vilh lime. They onIy relurned lo
re-lhyroidecomy vaIues vilh I-lhyroxine lrealmenl. AIlhough in olher secies,
incIuding humans, rimary hyolhyroidism may nol be associaled vilh
hyersomalolroism, lhe GH resonse lo surailuilary slimuIalion aears lo be
simiIar lo lhal in dogs. Of lhe hormones adminislered, nol onIy GHRH bul aIso
TRH, as demonslraled in Chaptcr 6, viII have increased GH reIease.
Irimary hyolhyroidism in humans may be associaled vilh
hyerroIaclinemia
27-30
and gaIaclorrhea.
30
GaIaclorrhea vas nol observed in lhe
beagIes vilh rimary hyolhyroidism, and lheir Iasma basaI IRL concenlralions
did nol increase afler induclion of hyolhyroidism. On lhe conlrary, Iasma IRL
concenlralions decreased significanlIy aboul 6 monlhs afler lhyroideclomy. OnIy
afler severaI monlhs of I-lhyroxine reIacemenl lheray did basaI Iasma IRL
concenlralions increase lovard re-lhyroideclomy IeveIs.
In aII dogs, lhe induclion of hyolhyroidism Ied lo graduaI increase in
iluilary size. Recenl MRI sludies of humans vilh rimary hyolhyroidism
reveaIed iluilary enIargemenl in 70% of cases.
30
L-lhyroxine lheray Ied lo
regression of lhe iluilary enIargemenl in aII lhree lrealed dogs, as has been
described in humans vilh iluilary enIargemenl secondary lo rimary
hyolhyroidism.
16,28,31

HislomorhoIogicaI and immunohislochemicaI sludies of lhe iluilary
gIand afler 3 years of rimary hyolhyroidism reveaIed massive lhyrolroe
hyerIasia, Iarge vacuoIaled lhyroid deficiency ceIIs, and decreased numbers of
mammolroes. The Iarge, aIe ceIIs, vilh abundanlIy vacuoIaled cyloIasm vere
simiIar lo lhe so-caIIed lhyroideclomy ceIIs or lhyroid deficiency ceIIs
described in hyolhyroid rals and humans vilh unlrealed rolracled rimary
hyolhyroidism.
27,32
The resence of miloses in severaI lhyrolroes suggesls lhal
lhe division of re-exisling lhyrolroes and/or Iess differenlialed (slem) ceIIs
conlribuled lo lhe deveIomenl of lhese lhyroid deficiency ceIIs. In addilion lo
lhis mechanism, lhe resuIls of doubIe immunoslaining indicaled lhal anolher
mechanism aIso vas oeralionaI. SeveraI ceIIs immunoslained for bolh GH and
TSH, simiIar lo lhe changes lhal have been reorled in hyolhyroid rals and
humans.
27,32
This has been inlerreled as lransilion of somalolroes inlo
lhyrolroes (lransdifferenlialion) and may exIain lhe aradoxicaI GH secrelion
observed afler slimuIalion vilh TRH in dogs vilh sonlaneous rimary
hyolhyroidism. The recenlIy idenlified muIlifunclionaI ceIIs lhal slore and secrele
lvo differenl adenohyohyseaI hormones (oIyhormonaI ceIIs) and/or resond lo
Chaler 9
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172

severaI reIeasing hormones may be invoIved in lransdifferenlialion and
aradoxicaI secrelion.
33

The observed decrease in basaI IRL concenlralions and lhe decrease in
IRL resonsiveness lo slimuIalion vilh lime may aIso be a resuIl of
lransdifferenlialion. In humans, Iong-slanding acromegaIy due lo ecloic
overroduclion of GHRH is associaled vilh somalolroe hyerIasia combined
vilh lhe aearance of mammosomalolroes, derived from Iaclolroes.
34
The
number of monohormonaI Iaclolroes vas very Iov in lhese alienls. Againsl lhe
background of lhese observalions, lhe graduaI decrease in Iaclolroe aclivily in our
dogs couId be due lo lhe recruilmenl of Iaclolroes lo form mammosomalolroes
and ossibIy lhyromammosomalolroes.
The resuIls of lhe sludy described in Chaptcr 7 demonslrale lhal lhe
abrul induclion of rimary hyolhyroidism Ied lo severaI funclionaI and
morhoIogicaI changes in lhe adenohyohysis. The main funclionaI changes
vere 1) graduaI Ioss of lhe TSH resonse lo lhe Iov circuIaling TT4 concenlralions,
beginning afler 4 monlhs, 2) graduaI deveIomenl of hyersecrelion of GH in lhe
firsl year, vhich lhen remained high, and 3) hyosecrelion of IRL afler 6 monlhs.
HislomorhoIogicaIIy, lhe enIarged iluilaries vere characlerized by lhyrolroe
hyerIasia, Iarge vacuoIaled lhyroid deficiency ceIIs, and doubIe-slaining ceIIs,
indicalive of lransdifferenlialion. These observalions exIain severaI of lhe earIier
observed biochemicaI fealures of canine hyolhyroidism.
The abrulIy induced form of hyolhyroidism in lhe beagIe dogs
described in Chaptcr 7 may nol be reresenlalive of lhe graduaIIy deveIoing
sonlaneous disease in dogs of olher breeds, ages, and gender. Therefore, lhe same
adenohyohyseaI funclion lesl as described in Chaptcr 7 vas erformed in 14
dogs vilh sonlaneous rimary hyolhyroidism and lhe resuIls vere comared
vilh lhose of 13 dogs vilh a Iov Iasma TT4

concenlralion bul no lhyroid Iesion,
i.e., dogs vilh non-lhyroidaI iIIness (Chaptcr 8). In lhe dogs vilh rimary
hyolhyroidism lhe funclion lesl vas reealed afler 4 and 12 veeks of subslilulion
lheray vilh I-lhyroxine.
As in lhe dogs vilh induced rimary hyolhyroidism, in lhe dogs vilh
sonlaneous rimary hyolhyroidism surailuilary slimuIalion did nol affecl
significanlIy lhe Iasma TSH concenlralions. WhiIe in bolh grous of dogs vilh
rimary hyolhyroidism (induced and sonlaneous) basaI Iasma TSH
concenlralions (furlher) decreased vilh I-lhyroxine lrealmenl, subslilulion lheray
did nol reslore TSH resonsiveness lo surailuilary slimuIalion.
asaI Iasma GH concenlralions in lhe dogs vilh sonlaneous rimary
hyolhyroidism vere significanlIy higher lhan lhose in lhe dogs vilh non-
lhyroidaI iIIness and increased furlher afler surailuilary slimuIalion. The Iover
Iasma GH concenlralion in lhe dogs vilh non-lhyroidaI iIIness increased afler
Summarizing discussion and concIusions
_________________________________________________________________________________

173

surailuilary slimuIalion, aIlhough Iess lhan in lhe dogs vilh rimary
hyolhyroidism. The higher GH resonse of lhe hyolhyroid dogs in comarison
vilh lhe dogs vilh non-lhyroidaI iIIness may have been caused by lhe aradoxicaI
resonse lo TRH in hyolhyroid dogs, as described in Chaptcr 6. In conlrasl, in lhe
dogs vilh non-lhyroidaI iIIness, lhere vas mosl IikeIy onIy a resonse lo GHRH.
As in lhe dogs vilh induced rimary hyolhyroidism, lhe Iasma GH
concenlralion decreased vilh I-lhyroxine lrealmenl in lhe dogs vilh sonlaneous
rimary hyolhyroidism.
There vas a remarkabIe difference vilh regard lo IRL belveen lhe dogs
vilh induced rimary hyolhyroidism and lhe dogs vilh sonlaneous rimary
hyolhyroidism. Afler 1-2 years, lhe dogs vilh induced rimary hyolhyroidism
had significanlIy Iover Iasma IRL concenlralions lhan iniliaIIy, and aIso lhe IRL
resonse lo surailuilary slimuIalion had decIined considerabIy. In conlrasl, in
lhe dogs vilh sonlaneous rimary hyolhyroidism lhe basaI IRL concenlralions
vere nol Iov, and in generaI lhere vas a good resonse lo surailuilary
slimuIalion, aIbeil vilh a vide varialion. This difference may in arl be due lo lhe
facl lhal lhe dogs vilh induced rimary hyolhyroidism vere sayed femaIes,
vhereas some of lhe dogs vilh sonlaneous rimary hyolhyroidism vere
sexuaIIy inlacl. In arlicuIar, lvo bilches vilh sonlaneous rimary
hyolhyroidism lhal had recenlIy slarled a nev eslrous cycIe had high basaI
Iasma IRL concenlralions vilh slrong resonsiveness lo surailuilary
slimuIalion.
The occurrence of hyerroIaclinemia and gaIaclorrhea has been
documenled earIier in a femaIe dog vilh rimary hyolhyroidism.
35
Aboul 8% of
humans vilh rimary hyolhyroidism have eIevaled IRL concenlralions vilh
gaIaclorrhea, vhich occurs mainIy in regnanl or Iaclaling vomen.
36
In Iine vilh
lhe occurrence of hyerroIaclinemia and gaIaclorrhea in humans vilh rimary
hyolhyroidism during regnancy and Iaclalion, in our sludy hyerroIaclinemia
vas observed rimariIy in lhe IuleaI hase of lhe eslrous cycIe of lhe dogs vilh
rimary hyolhyroidism. In dogs, rogeslerone in arlicuIar has a moduIalory
roIe in IRL secrelion.
37
The imorlance of lhe roIe of rogeslerone is consislenl
vilh lhe absence of hyerroIaclinemia and/or gaIaclorrhea in maIe dogs vilh
rimary hyolhyroidism. There is increasing evidence lhal in dogs il is nol an
increase in lhe Iasma rogeslerone concenlralion lhal slimuIales IRL reIease bul
ralher ils decrease. In nonregnanl bilches, lhe decrease in rogeslerone
concenlralion in lhe second arl of lhe IuleaI hase is associaled vilh an increase
in basaI Iasma IRL concenlralion.
37
AIso in seudoregnanl bilches, decreasing
Iasma rogeslerone concenlralions are associaled vilh increasing IRL reIease.
38

Moreover, adminislralion of a rogeslerone-recelor anlagonisl lo regnanl
bilches causes Iasma IRL concenlralions lo rise sharIy.
39
SimiIarIy in lhe lvo
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174

hyolhyroid bilches lhal recenlIy slarled a nev eslrous cycIe, lhe highesl basaI
IRL concenlralions and resonses lo slimuIalion occurred Iale in lhe IuleaI hase.
In lhe dogs vilh induced rimary hyolhyroidism, in lhe dogs vilh
sonlaneous rimary hyolhyroidism, and in lhe dogs vilh non-lhyroidaI iIIness,
basaI and slimuIaled concenlralions of LH and ACTH vere in aboul lhe same
range. L-lhyroxine subslilulion did nol affecl ACTH and LH IeveIs in lhe dogs
vilh induced rimary hyolhyroidism or in lhe dogs vilh sonlaneous rimary
hyolhyroidism.
The resuIls of lhe sludy described in Chaptcr 8 indicale lhal, simiIar lo
dogs vilh induced rimary hyolhyroidism, dogs vilh sonlaneous rimary
hyolhyroidism hyersecrele GH bul have IillIe or no TSH hyersecrelion. The
deveIomenl of hyerroIaclinemia (and ossibIe gaIaclorrhea) in rimary
hyolhyroidism seems lo occur onIy in sexuaIIy inlacl bilches. asaI and
slimuIaled Iasma adenohyohyseaI hormone concenlralions are nol aIlered in
dogs vilh non-lhyroidaI iIIness.

Irom lhe sludies described in lhis lhesis lhe foIIoving main concIusions can be
dravn:

In dogs vilh susecled hyolhyroidism, in vhich measuremenl of Iasma
concenlralions of lhyroxine (TT4 and/or fT4) and lhyroid-slimuIaling hormone
(TSH) rovides inconcIusive resuIls, measuremenl of lhyroid
99m
TcO4
-
ulake
can discriminale belveen rimary hyolhyroidism and non-lhyroidaI iIIness.

In heaIlhy dogs, TSH is secreled in a uIsaliIe manner vilh onIy smaII
fIuclualions. In dogs vilh rimary hyolhyroidism, lhe TSH uIses are more
ronounced and may occasionaIIy resuIl in Iasma TSH concenlralions vilhin
lhe reference range for eulhyroid dogs.

Irimary hyolhyroidism in dogs is associaled vilh eIevaled Iasma
concenlralions of grovlh hormone (GH) and of lhe grovlh faclor IGI-I. This
exIains some of lhe hysicaI changes of canine hyolhyroidism, mimicking lhe
syndrome of GH excess (acromegaIy).

In conlrasl lo heaIlhy dogs, dogs vilh rimary hyolhyroidism resond lo lhe
adminislralion of lhyrolroin-reIeasing hormone (TRH) vilh an increase in
Iasma GH concenlralions.



Summarizing discussion and concIusions
_________________________________________________________________________________

175

The iluilary funclion of dogs vilh rimary hyolhyroidism is characlerized
by:
- IniliaIIy high Iasma TSH concenlralions lhal decrease vilh lime lo IeveIs
lhal may faII vilhin lhe reference range for heaIlhy dogs. A graduaI Ioss of
TSH resonse lo surailuilary slimuIalion recedes lhis Iovering of basaI
Iasma TSH concenlralions.
- DeveIomenl of GH hyersecrelion, ersisling vilh lime.
- Hyosecrelion of roIaclin (IRL) in neulered bilches and IRL hyersecrelion
in sexuaIIy inlacl bilches during lhe IuleaI hase of lhe eslrus cycIe.

Iiluilary enIargemenl vilh lime is associaled vilh lhyrolroe hyerIasia,
Iarge vacuoIaled lhyroid deficiency ceIIs, and ceIIs doubIe-slaining for TSH
and GH.


Chaler 9
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176



1. eIshav I and Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Am Anim Hos Assoc 15: 17-23.
2. MiIIer A, NeIson RW, Scoll-Moncrieff }C, NeaI L and olloms GD 1992 SeriaI lhyroid
hormone concenlralions in heaIlhy eulhyroid dogs, dogs vilh hyolhyroidism, and
eulhyroid dogs vilh aloic dermalilis. r Vel } 148: 451-457.
3. Ramsey IK, Ivans H, Herrlage MI 1997 Thyroid-slimuIaling hormone and lolaI lhyroxine
concenlralions in eulhyroid, sick eulhyroid and hyolhyroid dogs. } SmaII Anim Iracl 38:
540-545.
4. Ielerson MI, MeIian C and NichoIs R 1997 Measuremenl of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroine concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Assoc 211: 1396-1402.
5. Dixon RM, Mooney CT 1999 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40: 72-78.
6. Kanlrovilz L, Ielerson MI, MeIian C, NichoIs R 2001 Serum lolaI lhyroxine, lolaI
lriiodolhyronine, free lhyroxine, and lhyrolrohin concenlralions in dogs vilh nonlhyroidaI
disease. } Am Vel Med Assoc 219: 765-769.
7. Kemainen R}, ehrend IN 2001 Diagnosis of canine hyolhyroidism. Ierseclives from a
lesling Iaboralory. Vel CIin Norlh Am SmaII Anim Iracl 31: 951-962.
8. }ensen AL, Iversen L, Hoier R, Krislensen I and Henriksen 1996 IvaIualion of an
immunoradiomelric assay for lhyrolroin in serum and Iasma samIes of dogs vilh
rimary hyolhyroidism. } Com IalhoI 114: 339-346.
9. runer }M, Scoll-Moncrieff CR, WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am
Vel Med Assoc. 212:1572-1575.
10. Scoll-Moncrieff }CR, NeIson RW, runer }M and WiIIiams DA 1998 Comarison of serum
concenlralions of lhyroid-slimuIaling hormone in heaIlhy dogs, hyolhyroid dogs, and
eulhyroid dogs vilh concurrenl disease } Am Vel Med Assoc 212: 387-391.
11. Mei| I, MoI }A and Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13: 465-468.
12. Mei| I, MoI }A, evers MM and Ri|nberk A 1997 ResiduaI iluilary funclion afler
lransshenoidaI hyohyseclomy in dogs vilh iluilary-deendenl hyeradrenocorlicism }
Indocr 155: 531-539.
13. Larsen IR, Davis TI, SchIumberger M-}, Hay ID 2003 Thyroid hysioIogy and diagnoslic
evaIualion of alienls vilh lhyroid disorders. In: WiIIiams Texlbook of IndocrinoIogy, 10
lh

ed. Larsen IR, Kronenberg HM, MeImed S, IoIonsky KS (Ids.) IhiIadeIhia, IA: W
Saunders. : 331-373.

Summarizing discussion and concIusions
_________________________________________________________________________________

177

14. Scoll-Moncrieff }CR and NeIson RW 1998 Changes in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolroin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and dogs vilh concurrenl disease. } Am Vel Med Assoc 213: 1435-
1438.
15. Shairo LI and Surks MI 2001 Hyolhyroidism. In: IrinciIes and Iraclice of IndocrinoIogy
and MelaboIism. 3
rd
ed. ecker KL (ed). IhiIadeIhia: Liincoll WiIIiams & WiIkins.: 445-
454.
16. SarIis N}, ruckner-Davis I, Doman }L, SkaruIis MC 1997 MRI-demonslrabIe regression of
a iluilary mass in a case of rimary hyolhyroidsm afler a veek of acule lhyroid hormone
lheray. } CIin IndocrinoI Melab 82: 808-811.
17. Sucuira MS, Camargo II, NickoIoff IL, AIderson IO, Wagner HN }r 1983 The roIe of
99m
Tc
erlechnelale ulake in lhe evaIualion of lhyroid funclion. Inl } NucI Med ioI 10: 29-33.
18. Na AMI, IoIIak YWIA, van den rom WI, Ri|nberk A 1994 Quanlilalive asecls of lhyroid
scinligrahy vilh erlechnelale (
99m
TcO4
-
) in cals. } Vel Inlern Med 8: 302-303.
19. Adams WH, DanieI G, Ielersen MG, Young K 1997 Quanlilalive
99m
Tc-erlechnelale lhyroid
scinligrahy in normaI beagIes. Vel RadioI UIlrasound 38: 323-328.
20. IeIdman and NeIson 1996. Hyolhyroidism. In: Canine and IeIine IndocrinoIogy and
Reroduclion. 2
nd
edilion. IeIdman IC and NeIson (Ids.). W Saunders Comany
IhiIadeIhia. I: 55-85.
21. Ri|nberk A 1996 Thyroids. In: CIinicaI IndocrinoIogy of Dogs and Cals. Ri|nberk A (ed).
Dordrechl, KIuver Academic IubIishers, : 35-39.
22. Iaggiano M, CriscuoIo T, Graziani M, Iorio S, Iisano G, Sinisi AA, Sinisi AM and eIIasleIIa A
1985 Iersilenl TRH-induced grovlh hormone reIease afler shorl-lerm and Iong-lerm L-
lhyroxine reIacemenl lheray in rimary congenilaI hyolhyroidis. CIin IndocrinoI 23:61-
66.
23. VareIa C, Cacicedo L, Iernandez G, de Ios IraiIes T and Sanchez Iranco I 1991 InfIuence of
hyolhyroidism duralion on deveIomenlaI changes in lhe hyolhaIamic faclors imIicaled
in grovlh hormone secrelion in lhe maIe ral. NeuroendocrinoIogy 54: 340-345.
24. Coiro V, VoIi R, Carelli L, Seroni G, Marchesi C, Vescovi II, Caffarri G, CoIIa R, Rossi G,
DavoIi C and Chiodera I 1994 InfIuence of lhyroid slalus on lhe aradoxicaI grovlh hormone
resonse lo lhyrolroin-reIeasing hormone in human obesily. MelaboIism 43: 514-517.
25. Hanev K, Ulsumi A, Sugavara A, Tanaka A, Iukazava H, Yoshida K and Abe K 1995
Inhanced Iasma GH resonses lo simuIlaneous adminislralion of TRH and GHRH in
alienls vilh rimary hyolhyroidism. IndocrinoI } 42: 43-47.
26. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IM and
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114: 275-282.


Chaler 9
_________________________________________________________________________________

178

27. VidaI S, Horvalh I, Kovacs K, Cohen SM, LIoyd RV, Scheilhauer W 2000.
Transdifferenlialion of somalolrohs lo lhyrolrohs in lhe iluilary of alienls vilh
rolracled rimary hyolhyroidism. Virchovs Arch 436: 43-51
28. VaIenla L}, Tamkin }, Soslrin R, IIias AN, Iisenberg H 1983 Regression of a iluilary
adenoma foIIoving Ievolhyroxine lheray of rimary hyolhyroidism. IerliI SleriI 40: 389-
392.
29. Dam|anovic S, Ioovic V, Ielakov M, D|urovic M, Dieguez C and Casanueva II 1996
Iiluilary enIargemenl due lo rimary hyolhyroidism: grovlh hormone resonse lo GHRH,
GHRI-6 and GHRH Ius GHRI-6. } Iedialr IndocrinoI Melab 9: 549-553.
30. Khava|a NM, Taher M, arham MI, Naser AA, Hadidy AM, Ahmad AT, Hamamy HA,
Yaghi NA, and A|Iouni KM. 2006 Iiluilary enIargemenl in alienls vilh hyolhyroidism
Indocr Iracl 12: 29-34.
31. ZeIissen IM}, Croughs R}M, Koerschaar HII, de ruin TW, Hendriks M}, van'l VerIaal
}W 1989 Irimary hyolhyroidism mimicking iluilary adenoma. Nelh } Med 35: 95-99.
32. Horvarlh I, LIoyd RV, Kovacs K 1990 IroyIlhiouracyI-induced hyolhyroidism resuIls in
reversibIe lransdifferenlialion for somalolrohs inlo lhyroideclomy ceIIs. A morhoIogic
sludy of lhe ral iluilary incIuding immunoeIeclron microscoy. Lab Invesl 63: 511-520.
33. ViIIaIobos C, Nuez L, Garcia-Sancho } 2004 Anlerior iluilary lhyrolroes are
muIlifunclionaI ceIIs Am } IhysioI IndocrinoI Melab 287: I1166-70.
34. VidaI S, Horvalh I, Kovacs K, LIoyd RV, Smylh HS. ReversibIe lransdifferenlialion:
Inlerconversion of somalolrohs and Iaclolrohs in iluilary hyerIasia. Mod IalhoI 2001,
14:20-28.
35. Corlese L, OIiva G, Verslegen }, CiarameIIa I, Iersechino A 1997 HyerroIaclinemia and
gaIaclorrhea associaled vilh rimary hyolhyroidism in a bilch. } SmaII Anim Iracl 38: 572-
575.
36. Raber W, GessI A, Novolny I, Vierhaer H 2003 HyerroIaclinaemia in hyolhyroidism:
cIinicaI significance and imacl of TSH normaIizalion. CIin IndocrinoI 58: 185-191.
37. Kooislra HD, Okkens AC 2002 Secrelion of grovlh hormone and roIaclin during
rogression of lhe IuleaI hase in heaIlhy dogs: a reviev. MoI CeII IndocrinoI 197: 167-172.
38. Okkens AC, DieIeman S}, Kooislra HS, evers MM 1997 IIasma concenlralions of roIaclin in
overlIy seudoregnanl Afghan hounds and lhe effecl of melergoIine. } Rerod IerliI, SuI
51: 295-301.
39. GaIac S, Kooislra HS, ulinar }, evers MM, DieIeman S}, Voorhoul G, Okkens AC 2000
Terminalion of midgeslalion regnancy in bilches vilh agIerislone, a rogeslerone recelor
anlagonisl. TheriogenoIogy 53: 941-950.



Chaptcr 10

5amcnvattcndc bcsprcking cn cnnc!usics
Chaler 10
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180
Samenvallende besreking en concIusies
_________________________________________________________________________________

181

i| de hond vordl de diagnose hyolhyreodie nogaI eens len onrechle
gesleId doordal elleIi|ke builen de schiIdkIieren geIegen facloren (bi|voorbeeId
medici|ngebruik en ziekle) kunnen Ieiden lol een Iage Iasmaconcenlralie van
lolaaI lhyroxine (TT4). In de |aren negenlig hoole men dal de invoering van
homoIoge immunoassays voor de beaIing van de Iasmaconcenlralie van
schiIdkIier-slimuIerend hormoon (TSH) bi| de hond de diagnosliek zou verbeleren.
De diagnose rimaire hyolhyreodie zou dan gesleId kunnen vorden o basis
van een Iage Iasma-TT4-concenlralie en een hoge Iasma-TSH-concenlralie.
Ichler, onderzoek, mel de TSH-slimuIalie lesl aIs gouden slandaard, Iiel zien dal
bi| een derde van de honden mel rimaire hyolhyreodie de Iasmaconcenlralie
van TSH niel verhoogd vas. Hel beIangri|ksle doeI van hel onderzoek beschreven
in dil roefschrifl vas om de hyolhese le loelsen dal rimaire hyolhyreodie bi|
de hond Ieidl lol veranderingen in de adenohyofyse die hel olreden van Iage
Iasmaconcenlralies van TSH kunnen verkIaren. Daarnaasl verd gehool dal hel
onderzoek ook belrouvbaardere melhodes voor de diagnose rimaire
hyolhyreodie bi| de hond zou oIeveren.
ZoaIs in hel voorgaande reeds is aangegeven, kunnen Iasma-TT4-
concenlralies Iaag zi|n bi| honden mel non-lhyroidaI iIIness (NTI), en daardoor
kan hel moeiIi|k zi|n om onderscheid le maken lussen hyolhyreodie en NTI bi| de
hond. HnnIdstuk 3 beschri|fl de bevindingen mel een grool aanlaI diagnoslische
leslen bi| 30 honden mel een Iage Iasma-TT4-concenlralie. De definilieve diagnose
rimaire hyolhyreodie of NTI beruslle bi| deze honden o hisloIogisch
onderzoek van een schiIdkIierbiol.
AIIe honden mel rimaire hyolhyreodie hadden zeer Iage Iasma-TT4-
concenlralies. Dil koml overeen mel de bevindingen in eerdere sludies die Iielen
zien dal de beaIing van de Iasma-TT4-concenlralie een gevoeIige lesl is voor de
diagnose hyolhyreodie.
1-5
De honden mel NTI hadden een Iasma-TT4-
concenlralie die significanl hoger vas dan die van de honden mel rimaire
hyolhyreodie, maar enkeIe honden mel NTI hadden ook een zeer Iage Iasma-
TT4-concenlralie. Dil bevesligl eerdere vaarnemingen dal de hoge sensilivileil van
TT4-beaIingen bi| de diagnosliek van hyolhyreodie heIaas geaard gaal mel een
Iage secificileil.
5,6

De serumconcenlralies van vri| T4 (fT4) van de honden mel rimaire
hyolhyreodie varen significanl Iager dan die van de honden mel NTI. AIIe
honden mel hyolhyreodie, mel uilzondering van 1 hond, hadden een serum-fT4-
concenlralie onder de deleclieIimiel van de assay. Daarnaasl vas er maar 1 hond
mel NTI die een serumconcenlralie van fT4 onder de deleclieIimiel van de assay
had. De gering belere secificileil van fT4 vergeIeken mel die van TT4 kan een
reden zi|n om deze lesl roulinemalig le gaan gebruiken bi| de diagnosliek van
hyolhyreodie bi| de hond.
4,7

Chaler 10
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182

De sli|ging van de Iasmaconcenlralie van TT4 na slimuIalie mel TSH vas
significanl groler bi| de honden mel NTI dan bi| de honden mel rimaire
hyolhyreodie. i| enkeIe honden mel NTI vas er echler maar een minimaIe
loename in de Iasma-TT4-concenlralie. Indien eerder vaslgesleIde afkavaarden
voor hyolhyreodie zouden zi|n loegeasl (na TSH slimuIalie een
Iasmaconcenlralie van TT4 >25 nmoI/I en/of een Iasmaconcenlralie van TT4 die
minslens 1,5 keer de basaIe TT4 concenlralie bedraagl),
4
dan zouden enkeIe honden
mel NTI on|uisl gecIassificeerd zi|n aIs honden mel rimaire hyolhyreodie. Hel is
vaarschi|nIi|k dal deze miscIassificalie ook is ogelreden bi| eerdere onderzoeken
naar hyolhyreodie bi| de hond, vaarbi| de TSH-slimuIalielesl aIs gouden
slandaard is gebruikl om andere leslen le evaIueren voor de diagnosliek van
hyolhyreodie bi| de hond. Niellemin mag de TSH-slimuIalielesl nog sleeds
vorden beschouvd aIs de minsl sIechle biochemische schiIdkIierfunclielesl voor
hel onderscheid lussen rimaire hyolhyreodie en NTI bi| de hond.
Ien reIalief grool aanlaI honden mel rimaire hyolhyreodie (6 van de 14)
had een Iasma-TSH-concenlralie die binnen hel referenliegebied voor eulhyreole
honden Iag. Dil resuIlaal koml overeen mel eerdere vaarnemingen (mel de TSH-
slimuIalielesl aIs de gouden slandaard) dal bi| een derde van de honden mel
rimaire hyolhyreodie de Iasma-TSH-concenlralie niel verhoogd is.
3,4,5,8,9,10
De
basaIe Iasma-TSH-concenlralies bi| honden mel NTI vieIen binnen hel
referenliegebied, mel uilzondering van een hond die een verhoogde Iasma-TSH-
concenlralie had. Hel voorkomen van Iasma-TSH-concenlralies hoger dan hel
referenliegebied is eerder beschreven bi| eulhyreole honden.
3,4,5,10

De beaIing van de TSH-resons na loediening van TRH biedl een sneIIe,
veiIige en effeclieve melhode om de reservecaacileil van de lhyreolroe ceIIen in
de hyofyse le onderzoeken.
11,12
De TRH-slimuIalielesl zou mogeIi|k ook gebruikl
kunnen vorden bi| de diagnosliek van rimaire hyolhyreodie, vaarbi|
loediening van TRH o lheorelische gronden dan zou moelen Ieiden lol een
grolere dan normaIe TSH-afgifle.
13
Ichler bi| de honden mel rimaire
hyolhyreodie vas er geen overmalige TSH-resons na slimuIalie mel TRH.
InlegendeeI, in overeenslemming mel de resuIlalen van eerdere onderzoeken
3,14

bIeek de TSH-sli|ging significanl geringer dan bi| de honden mel NTI. Daarnaasl
Ialen de resuIlalen van hel onderzoek beschreven in HnnIdstuk 6 ook zien dal
loediening van TRH een significanle sli|ging van de Iasma-TSH-concenlralie
veroorzaakl bi| gezonde conlroIehonden maar niel bi| honden mel rimaire
hyolhyreodie. Dus de hyofysaire TSH-resons o TRH is bi| honden mel
rimaire hyolhyreodie aanzienIi|k verminderd. Dil is in legensleIIing lol de
silualie bi| mensen mel rimaire hyolhyreodie vaar de TSH-resons o TRH
aIli|d is loegenomen.
15
ZeIfs humane alinlen mel Iangdurige rimaire
hyolhyreodie en hyerIasie van de lhyreolroe ceIIen reageren nog overmalig
Samenvallende besreking en concIusies
_________________________________________________________________________________

183

o TRH-slimuIalie.
16
Dus een afvezige of verminderde TSH-resons bi| een TRH-
slimuIalielesl is secifiek voor honden mel hyolhyreodie en vooraIsnog
onverkIaard.
Derlig minulen na TRH-loediening vas er geen significanl verschiI lussen
de Iasma-TSH-concenlralies van honden mel hyolhyreodie en honden mel NTI.
ZoaIs eerder beschreven, is dus hel gebruik van de TRH-slimuIalielesl, mel
beaIing van Iasma-TSH-concenlralies, niel geschikl voor hel onderscheid lussen
rimaire hyolhyreodie en NTI.
13,14

De berekening van de rocenlueIe oname van
99m
TcO4
-
in hel
schiIdkIierveefseI is zeer bruikbaar gebIeken bi| de beoordeIing van de
schiIdkIierfunclie bi| mensen en kallen.
17,18
Dil goId ook voor de honden mel
hyolhyreodie in dil onderzoek. Ir vas nameIi|k geen overIaing van de
rocenlueIe
99m
TcO4
-
-oname o 45 lol 120 minulen na de in|eclie van hel
radioaclieve isoloo lussen de honden mel rimaire hyolhyreodie en de honden
mel NTI. De
99m
TcO4
-
-oname bi| de honden mel NTI vas vergeIi|kbaar mel de
oname die beschreven is bi| gezonde beagIes.
19

Samenvallend bevesligen de resuIlalen van hel onderzoek beschreven in
HnnIdstuk 3 eerdere vaarnemingen dal de biochemische diagnose van rimaire
hyolhyreodie niel eenvoudig is en geaard gaal mel secifieke robIemen door
de overIaingen van de bIoedconcenlralies van TT4, fT4 en TSH bi| honden mel
hyolhyreodie en honden mel NTI. Toeassing van eerder vaslgesleIde
diagnoslische crileria bi| gebruik van de TSH-slimuIalielesl bIeek ook niel aIli|d lol
een |uisle diagnose le Ieiden. ovendien overIalen de resuIlalen van deze lesl bi|
de 2 groeen honden. In eerdere onderzoeken, vaarbi| de TSH-slimuIalielesl aIs
gouden slandaard voor de diagnose van hyolhyreodie verd gebruikl, is mogeIi|k
srake geveesl van miscIassificalie. De kvanlilalieve meling van de oname in de
schiIdkIieren van
99m
TcO4
-
had hel hoogsle onderscheidend vermogen in de
differenlialie voor hyolhyreodie en NTI.
Om le onderzoeken of Iasma-TSH-concenlralies binnen hel
referenliegebied, zoaIs die bi| sommige honden mel rimaire hyolhyreodie
vorden gezien, verkIaard kunnen vorden door fIuclualies in de hyofysaire
afgifle van TSH, verd bi| 7 beagIe leven, zoveI voor aIs na de induclie van
rimaire hyolhyreodie, hel Iasma-TSH-rofieI vaslgesleId door gedurende 6
uur eIke 10 minulen een bIoedmonsler le nemen en in deze monslers de Iasma-
TSH-concenlralie le melen (HnnIdstuk 4). Ir vas duideIi|ke uIsalieIe secrelie van
TSH bi| de honden li|dens hyolhyreodie en li|dens eulhyreodie varen er sIechls
kIeine fIuclualies in de Iasma-TSH-concenlralie. De fIuclualies rond de TSH-
basaaIvaarden bi| honden mel genduceerde rimaire hyolhyreodie kunnen
deeIs verkIaren vaarom bi| sommige honden mel sonlane rimaire
hyolhyreodie de Iasma-TSH-concenlralies binnen hel referenliegebied van
Chaler 10
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184

eulhyreole honden kunnen vaIIen. Omdal geraorleerd is dal de gemiddeIde
Iasma-TSH-concenlralie van honden mel sonlane rimaire hyolhyreodie
sIechls marginaaI hoger is dan de bovengrens van hel referenliegebied van TSH bi|
eulhyreole honden,
4,10
zouden de fIuclualies rond de TSH-basaaIvaarden o dil
niveau kunnen resuIleren in Iasmaconcenlralies binnen hel referenliegebied van
eulhyreole honden.
De verschi|nseIen van rimaire hyolhyreodie bi| honden zi|n ondermeer
gevichlsloename, Ielhargie en dermaloIogische veranderingen zoaIs kaaIheid en
huidverdikkingen door ohoing van mucooIysacchariden.
20,21
Om de mogeIi|ke
belrokkenheid van een veranderende afgifle van groeihormoon (GH) le leslen bi|
honden mel rimaire hyolhyreodie verden bi| 7 beagIes de 6-uurs uIsalieIe
secreliealronen van GH onderzochl zoveI voor aIs na induclie van
hyolhyreodie (HnnIdstuk 5). In legensleIIing lol helgeen geraorleerd is bi|
verscheidene andere zoogdieren, bIeken honden mel hyolhyreodie een
verhoogde GH-secrelie le hebben. De verhoogde GH-secrelie heefl ook endocriene
reIevanlie zoaIs bIi|kl uil de verhoogde Iasmaconcenlralies van insuIine-achlige
groeifaclor-I (IGI-I) en enkeIe fysieke veranderingen die assen bi| acromegaIie.
i| eulhyreole mensen, rallen en honden Ieidl de loediening van TRH niel
lol vaarneembare veranderingen in de Iasmaconcenlralie van GH.
22-26
Ichler, de
loediening van TRH Ieidl bi| mensen en rallen mel rimaire hyolhyreodie veI lol
een significanle loename van de Iasmaconcenlralie van GH.
22-25
Om le
onderzoeken of loediening van TRH bi| honden mel rimaire hyolhyreodie ook
Ieidl lol een aradoxaIe sli|ging van de Iasma-GH-concenlralie, verd hel effecl
onderzochl van de inlraveneuze loediening van TRH o de GH- secrelie bi| 10
honden mel rimaire hyolhyreodie en 6 gezonde conlroIehonden (HnnIdstuk 6).
i| de honden mel rimaire hyolhyreodie sleeg de Iasma-GH-concenlralie
significanl na loediening van TRH. ZoaIs eerder ook geraorleerd door Rulleman
.
26
, Ieidde TRH- loediening niel lol een significanle sli|ging van de Iasma-GH-
concenlralie bi| de gezonde conlroIehonden.
i| mensen mel Iangdurige rimaire hyolhyreodie vordl uilgebreide
hyerIasie van lhyreolroe ceIIen gezien. DubbeIe immunokIeuring, immuno-
eIeclronenmicroscoie en in silu hybridalie loonde de aanvezigheid aan van
bihormonaIe ceIIen, ofleveI ceIIen die zoveI GH aIs TSH roduceren.
27
Dus naasl
hyerIasie van lhyreolroe ceIIen, Ii|kl ook lransdifferenlialie van somalolroe
ceIIen naar lhyreolroe ceIIen bi| le dragen aan de loename van TSH-roducerende
ceIIen.
Om le onderzoeken of dergeIi|ke morfoIogische veranderingen ook
olreden in de hyofyse van honden mel rimaire hyolhyreodie, verden 7
beagIes gevoIgd gedurende 3 |aar na de induclie van rimaire hyolhyreodie

Samenvallende besreking en concIusies
_________________________________________________________________________________

185

(HnnIdstuk 7). Drie van deze honden verden nog vervoIgd voor een addilioneIe
eriode van 1,5 |aar lervi|I ze behandeId verden mel I-lhyroxine. IIke 2 maanden
verd de adenohyofysaire funclie onderzochl door middeI van een
gecombineerde in|eclie van vier hyolhaIame hormonen (CRH, GHRH, GnRH en
TRH) en de beaIing van de Iasmaconcenlralies van de hyofysaire hormonen
(ACTH, GH, LH, IRL en TSH). Daarnaasl verd iedere 6 maanden de hyofyse in
beeId gebrachl mel behuI van comulerlomografie (CT). i| zoveI de 4 honden
mel onbehandeIde hyolhyreodie aIs bi| de 3 honden mel hyolhyreodie en
aansIuilende behandeIing mel I-lhyroxine verden hislomorfoIogische en
immunohislochemische sludies van de hyofyse verrichl.
De induclie van rimaire hyolhyreodie Ieidde lol een sli|ging van de
Iasma-TSH-concenlralie mel een faclor 15. De Iasma-TSH-concenlralies bIeven
sIechls een aanlaI maanden zo hoog. Daarna lrad een geIeideIi|ke daIing in.
Gedurende hel groolsle deeI van de observalieeriode bIeven de Iasma-TSH-
concenlralies echler veI significanl hoger dan de iniliIe vaarden voorafgaande
aan de induclie van hyolhyreodie. AIIeen o hel einde van dil deeI van de sludie
(ca. 3 |aar na induclie van rimaire hyolhyreodie) varen de Iasma-TSH-
concenlralies niel meer significanl verschiIIend van de iniliIe vaarden. De TSH-
resons na surahyofysaire slimuIalie nam nog sneIIer af. Deze vaarnemingen
kunnen de vaak niel verhoogde basaIe Iasma-TSH-concenlralies bi| honden mel
sonlane rimaire hyolhyreodie verkIaren.
asaIe Iasma-GH-concenlralies namen geIeideIi|k loe gedurende hel
eersle |aar en bIeven daarna o dil verhoogde niveau. Dus in legensleIIing lol de
basaIe TSH-concenlralie nam de basaIe GH-concenlralie niel af in de Ioo van de
li|d. AIIeen na behandeIing mel I-lhyroxine keerden deze vaarden lerug lol de
vaarden zoaIs aanvezig voor de induclie van hyolhyreodie. HoeveI bi| andere
diersoorlen en de mens rimaire hyolhyreodie niel geaard gaal mel
hyersomalolroisme, is de aradoxaIe GH-resons na surahyofysaire
slimuIalie veI vergeIi|kbaar mel die bi| honden. Van de loegediende hormonen
kunnen zoveI GHRH aIs TRH de GH afgifle hebben verhoogd (zie ook
HnnIdstuk 6).
Irimaire hyolhreodie bi| de mens kan geaard gaan mel
hyerroIaclinemie
27-30
en gaIaclorrhoea.
30
GaIaclorrhoea verd niel vaargenomen
bi| de beagIes mel rimaire hyolhyreodie. Ook namen hun basaIe Iasma-IRL-
concenlralies niel loe na de induclie van hyolhyreodie. InlegendeeI, de Iasma-
IRL-concenlralies namen significanl af vanaf ongeveer 6 maanden na de induclie
van hyolhyreodie. Ias na een aanlaI maanden behandeIen mel I-lhyroxine
namen de basaIe Iasma-IRL-concenlralies loe lol de vaarden voor de induclie
van hyolhyreodie.

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186

i| aIIe honden Ieidde de induclie van hyolhyreodie lol een geIeideIi|ke
loename van de groolle van de hyofyse. In een recenle ubIicalie is le Iezen dal
mel MRI ook bi| 70% van mensen mel hyolhyreodie een hyofysevergroling
aanloonbaar is.
30
ehandeIing mel I-lhyroxine Ieidde lol regressie van de
hyofysevergroling bi| de drie behandeIde honden, zoaIs ook beschreven is bi|
mensen mel een hyofysevergroling lengevoIge van rimaire hyolhyreodie.
16,28,31

HislomorfoIogisch en immunohislochemisch onderzoek van de hyofyse
na 3 |aar (onbehandeIde) rimaire hyolhyreodie loonde een uilgebreide
hyerIasie van lhyreolroe ceIIen, een grool aanlaI gevacuoIiseerde, bIeke ceIIen,
en een afgenomen aanlaI mammolroe ceIIen. De grole bIeke ceIIen mel
gevacuoIiseerd cyloIasma verloonden slerke geIi|kenis mel de zogenaamde
lhyroideclomy ceIIs of lhyroid deficiency ceIIs, zoaIs beschreven bi|
hyolhyreole rallen en mensen mel een aI Ianger beslaande onbehandeIde
rimaire hyolhyreodie.
27,32
De aanvezigheid van milosen in verscheidende
lhyreolroe ceIIen suggereerl dal de deIing van rexislenle lhyreolroe ceIIen
en/of minder gedifferenlieerde (slam)ceIIen heefl bi|gedragen aan de onlvikkeIing
van de lhyroid deficiency ceIIs. De resuIlalen van een dubbeIe immunokIeuring
lonen dal er ook nog een ander mechanisme oeralioneeI vas. Meerdere ceIIen
bIeken immunoosilief voor zoveI GH aIs TSH, vergeIi|kbaar mel de
veranderingen die zi|n geraorleerd bi| hyolhyreole rallen en mensen.
27,32
Deze
dubbeIkIeuring asl bi| de overgang van somalolroe naar lhyreolroe ceIIen
(lransdifferenlialie) en kan de aradoxaIe GH secrelie na slimuIalie mel TRH bi|
honden mel sonlane rimaire hyolhyreodie verkIaren. De recenl
gedenlificeerde muIlifunclioneIe ceIIen die lvee verschiIIende adenohyofysaire
hormonen bevallen en afscheiden (oIyhormonaIe ceIIen) en/of reageren o
meerdere hyolhaIame hormonen zouden aan de basis kunnen slaan van de
lransdifferenlialie en de aradoxaIe secrelie.
33

De vaargenomen afname van de basaIe Iasma-IRL-concenlralies en de
verminderde IRL-resons o slimuIalie in de Ioo van de li|d kunnen ook een
gevoIg zi|n van lransdifferenlialie. i| de mens gaal Iangdurige acromegaIie aIs
gevoIg van ecloische overroduclie van GHRH geaard mel hyerIasie van
somalolroe ceIIen en hel verschi|nen van mammasomalolroe ceIIen (onlslaan uil
Iaclolroe ceIIen).
34
Hel aanlaI monohormonaIe Iaclolroe ceIIen vas erg Iaag bi|
deze alinlen. Tegen de achlergrond van deze vaarnemingen kan de geIeideIi|ke
afname van de Iaclolroe aclivileil bi| onze honden hel gevoIg zi|n van de
rekrulering van Iaclolroe ceIIen voor de vorming van mammosomalolroe en
mogeIi|k lhyromammosomalolroe ceIIen.
Samenvallend lonen de resuIlalen van de sludies die beschreven zi|n in
HnnIdstuk 7 aan dal de abrule induclie van rimaire hyolhyreodie Ieidl lol
verscheidene funclioneIe en morfoIogische veranderingen in de adenohyofyse.
Samenvallende besreking en concIusies
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187

De beIangri|ksle funclioneIe veranderingen zi|n: 1) een geIeideIi|k olredend
verIies van de TSH-resonsivileil o de Iage Iasmaconcenlralies van
schiIdkIierhormoon, een roces dal beginl ongeveer 4 maanden na hel onlslaan
van de hyolhyreodie, 2) geIeideIi|ke onlvikkeIing van hyersecrelie van GH in
hel eersle |aar, vaarna de Iasma-GH-concenlralies conlinu hoog bIi|ven, en 3)
hyosecrelie van IRL vanaf 6 maanden. HislomorfoIogisch verden de vergrole
hyofyses gekarakleriseerd door lhyreolroe hyerIasie, grole gevacuoIiseerde
lhyroid deficiency ceIIs, en dubbeIkIeurende ceIIen. Dil zi|n slerke aanvi|zingen
voor lransdifferenlialie. Deze vaarnemingen verkIaren verscheidene van de eerder
vaargenomen biochemische kenmerken van hyolhyreodie bi| de hond.
De abrul genduceerde vorm van hyolhyreodie bi| de beagIes
beschreven in HnnIdstuk 7 is misschien niel reresenlalief voor de geIeideIi|ke
onlvikkeIing van de sonlane ziekle, en mogeIi|k ook afhankeIi|k van ras, Ieefli|d
en gesIachl. Daarom verd dezeIfde funclielesl van de adenohyofyse uilgevoerd
bi| 14 honden mel sonlane hyolhyreodie. De resuIlalen verden vergeIeken mel
die van 13 honden mel een Iaag Iasma TT4 concenlralie zonder schiIdkIierIaesie
(honden mel NTI) (HnnIdstuk 8). i| de honden mel rimaire hyolhyreodie
verd de funclielesl herhaaId na 4 en 12 veken van subslilulielheraie mel I-
lhyroxine.
ZoveI bi| de honden mel exerimenleeI ogevekle rimaire
hyolhyreodie, aIs bi| de honden mel sonlane rimaire hyolhyreodie, vas er
geen significanl effecl van surahyofysaire slimuIalie o de Iasma-TSH-
concenlralie. Tervi|I in beide groeen van honden mel rimaire hyolhyreodie
(genduceerd en sonlaan) de basaIe Iasma-TSH-concenlralies (verder) afnamen
mel I-lhyroxine behandeIing, verbelerde mel de subslilulielheraie de TSH-
resons o surahyofysaire slimuIalie niel.
De basaIe Iasma-GH-concenlralies van de honden mel sonlane rimaire
hyolhyreodie varen significanl hoger dan die van de honden mel NTI en ze
namen verder loe na surahyofysaire slimuIalie. De Iagere Iasma-GH-
concenlralies bi| de honden mel NTI namen ook loe na surahyofysaire
slimuIalie, echler minder dan de sli|ging die gezien verd bi| de honden mel
rimaire hyolhyreodie. De hogere GH-resons bi| de honden mel
hyolhyreodie in vergeIi|king mel die bi| de honden mel NTI zou hel gevoIg
kunnen zi|n van de aradoxaIe resons o TRH bi| de honden mel hyolhyreodie,
zoaIs beschreven in HnnIdstuk 6. i| de honden mel NTI vas er vaarschi|nIi|k
aIIeen een resons o GHRH. ZoveI bi| de honden mel exerimenleeI ogevekle
rimaire hyolhyreodie aIs bi| de honden mel sonlane rimaire hyolhyreoidie
nam de Iasma-GH-concenlralie af na behandeIing mel I-lhyroxine.
Ir vas een omerkeIi|k verschiI lussen honden mel genduceerde rimaire
hyolhyreodie en honden mel sonlane rimaire hyolhyreodie mel belrekking
Chaler 10
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188

lol IRL. Na 1-2 |aar hadden de honden mel genduceerde rimaire hyolhyreodie
een significanl Iagere Iasma-IRL-concenlralie dan inilieeI en ook de IRL resons
na surahyofysaire slimuIalie vas aanzienIi|k afgenomen. Daarenlegen hadden
de honden mel sonlane rimaire hyolhyreodie geen Iage basaIe Iasma-IRL-
concenlralies en in hel aIgemeen vas er een goede resons o surahyofysaire
slimuIalie. Dil verschiI zou gedeeIleIi|k veroorzaakl kunnen vorden door hel feil
dal de honden mel genduceerde rimaire hyolhyreodie gecaslreerde leven
varen, lervi|I sommige honden mel sonlane rimaire hyolhyreodie seksueeI
inlacl varen. Mel name 2 leven mel sonlane rimaire hyolhyreodie die nel een
nieuve oeslrische cycIus varen geslarl hadden hoge Iasma-IRL-concenlralies
mel een slerke reaclie o surahyofysaire slimuIalie.
Hel olreden van hyerroIaclinemie en gaIaclorrhoea is eerder
gedocumenleerd bi| een leef mel rimaire hyolhyreodie.
35
Ongeveer 8% van de
mensen mel rimaire hyolhyreodie hebben verhoogde IRL-concenlralies mel
gaIaclorrhoea, dil vordl vooraI gezien bi| zvangere en Iaclerende vrouven.
36

VergeIi|kbaar mel hel olreden van hyerroIaclinemie en gaIaclorrhoea bi|
mensen mel rimaire hyolhyreodie li|dens de zvangerscha en de Iaclalie, verd
de hyerroIaclinemie bi| honden mel sonlane hyolhyreodie vooraI gezien
li|dens de IuleaIe fase van de oeslrische cycIus.
i| de hond heefl rogesleron een slerk moduIerende effecl o de IRL
secrelie.
37
Hel beIang van rogesleron bIi|kl uil de afvezigheid van
hyerroIaclinemie en/of gaIaclorrrhea bi| reuen mel rimaire hyolhyreodie. Ir
is sleeds meer bevi|s dal hel bi| honden niel de loename van de
rogesleronconcenlralie is die de IRL-secrelie slimuIeerl, maar eerder de afname
van de rogesleronconcenlralie in Iasma. i| niel-drachlige leven gaal de afname
van de rogesleronconcenlralie in hel lveede deeI van de IuleaIe fase geaard mel
een loename van de basaIe IRL-concenlralie.
37
Ook bi| schi|nzvangere leven gaal
een daIing van de Iasma-rogesleron-concenlralie geaard mel een loename van
de IRL-afgifle.
38
ovendien Ieidl loediening van een rogesleron-recelor-
anlagonisl aan drachlige leven lol een slerke sli|ging van de IRL-concenlralie.
39
i|
de 2 hyolhyreole leven in ons onderzoek die recenl een nieuve oeslrische cycIus
varen geslarl lraden de hoogsle basaIe Iasma-IRL-concenlralies en de slerksle
reaclie o slimuIalie o in de Iaal-IuleaIe fase.
i| de honden mel genduceerde rimaire hyolhyreodie, de honden mel
sonlane rimaire hyolhyreodie, en de honden mel NTI varen de basaIe en
geslimuIeerde Iasmaconcenlralies van LH en ACTH min of meer vergeIi|kbaar.
i| zoveI de honden mel genduceerde rimaire hyolhyreodie aIs bi| de honden
mel sonlane rimaire hyolhyreodie veranderden deze vaarden niel na I-
lhyroxine-subslilulie.

Samenvallende besreking en concIusies
_________________________________________________________________________________

189

Samenvallend lonen de resuIlalen van de sludie beschreven in HnnIdstuk
8 dal zoveI honden mel exerimenleeI ogevekle rimaire hyolhyreodie aIs
honden mel sonlane rimaire hyolhyreodie een hyersecrelie van GH hebben,
maar reIalief veinig hyersecrelie van TSH. De onlvikkeIing van
hyerroIaclinemie (en mogeIi|k gaIaclorrhea) Ii|kl aIIeen o le lreden bi|
ongecaslreerde leven mel rimaire hyolhyreodie. asaIe en geslimuIeerde
Iasmaconcenlralies van de adenohyofysaire hormonen zi|n niel veranderd bi|
honden mel NTI.

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190

De resuIlalen van hel onderzoek beschreven in dil roefschrifl maken de voIgende
concIusies mogeIi|k:

i| honden die verdachl vorden van hyolhyreodie, maar vaarbi| de
Iasmaconcenlralies van lhyroxine (TT4 en/of fT4) en schiIdkIier-
slimuIerend hormoon (TSH) niel lol een diagnose Ieiden, kan mel de
meling van de oname van
99m
TcO4
-
in de schiIdkIieren onderscheid
vorden gemaakl lussen rimaire hyolhyreodie en NTI.

i| gezonde honden vordl TSH o een uIsalieIe manier afgegeven door
de hyofyse, zi| hel mel sIechls kIeine fIuclualies. i| honden mel rimaire
hyolhyreodie zi|n de TSH-uIsen meer uilgesroken en kan de
uIsalieIe TSH-secrelie soms Ieiden lol Iasma-TSH-concenlralies binnen
de referenlievaarden voor eulhyreole honden.

Irimaire hyolhyreodie bi| honden gaal samen mel verhoogde
Iasmaconcenlralies van groeihormoon (GH) en de groeifaclor IGI-1. Dil
verkIaarl vaarom de fysieke veranderingen van honden mel
hyolhyreodie zoveeI geIi|kenis verlonen mel de veranderingen bi|
acromegaIie.

In legensleIIing lol de silualie bi| gezonde honden heefl de loediening van
TRH bi| honden mel hyolhyreodie een sli|ging van de Iasma-GH-
concenlralie lol gevoIg.

De hyofysefunclie van honden mel rimaire hyolhyreodie kan aIs
voIgl omschreven vorden:
- IniliIe hoge Iasma-TSH-concenlralies die mel de li|d kunnen afnemen
lol vaarden die binnen hel referenliegebied Iiggen voor gezonde honden.
De verIaging van de basaIe Iasma-TSH-concenlralies vordl voorafgegaan
door een geIeideIi|k verIies van de TSH-resons o surahyofysaire
slimuIalie
- Hyersecrelie van GH, die ersisleerl in de li|d.
- Hyosecrelie van IRL bi| gecaslreerde leven en hyersecrelie van IRL
bi| ongecaslreerde leven li|dens de IuleaIe fase van de oeslrische cycIus.

De groolle van de hyofyse van honden mel rimaire hyolhyreodie
neeml loe in de Ioo van de li|d en dil gaal geaard mel hyerIasie van
lhyrolroe ceIIen, grole gevacuoIiseerde lhyroid deficiency ceIIen, en
ceIIen die dubbeIkIeuren voor TSH en GH.
Samenvallende besreking en concIusies
_________________________________________________________________________________

191

$

1. eIshav I and Ri|nberk A 1979 Radioimmunoassay of Iasma T4 and T3 in lhe diagnosis of
rimary hyolhyroidism in dogs. } Am Anim Hos Assoc 15: 17-23.
2. MiIIer A, NeIson RW, Scoll-Moncrieff }C, NeaI L and olloms GD 1992 SeriaI lhyroid
hormone concenlralions in heaIlhy eulhyroid dogs, dogs vilh hyolhyroidism, and
eulhyroid dogs vilh aloic dermalilis. r Vel } 148: 451-457.
3. Ramsey IK, Ivans H, Herrlage MI 1997 Thyroid-slimuIaling hormone and lolaI lhyroxine
concenlralions in eulhyroid, sick eulhyroid and hyolhyroid dogs. } SmaII Anim Iracl 38:
540-545.
4. Ielerson MI, MeIian C and NichoIs R 1997 Measuremenl of serum lolaI lhyroxine,
lriiodolhyronine, free lhyroxine, and lhyrolroine concenlralions for diagnosis of
hyolhyroidism in dogs. } Am Vel Med Assoc 211: 1396-1402.
5. Dixon RM, Mooney CT 1999 IvaIualion of serum free lhyroxine and lhyrolroin
concenlralions in lhe diagnosis of canine hyolhyroidism. } SmaII Anim Iracl 40: 72-78.
6. Kanlrovilz L, Ielerson MI, MeIian C, NichoIs R 2001 Serum lolaI lhyroxine, lolaI
lriiodolhyronine, free lhyroxine, and lhyrolrohin concenlralions in dogs vilh nonlhyroidaI
disease. } Am Vel Med Assoc 219: 765-769.
7. Kemainen R}, ehrend IN 2001 Diagnosis of canine hyolhyroidism. Ierseclives from a
lesling Iaboralory. Vel CIin Norlh Am SmaII Anim Iracl 31: 951-962.
8. }ensen AL, Iversen L, Hoier R, Krislensen I and Henriksen 1996 IvaIualion of an
immunoradiomelric assay for lhyrolroin in serum and Iasma samIes of dogs vilh
rimary hyolhyroidism. } Com IalhoI 114: 339-346.
9. runer }M, Scoll-Moncrieff CR, WiIIiams DA 1998 Iffecl of lime of samIe coIIeclion on
serum lhyroid-slimuIaling hormone concenlralions in eulhyroid and hyolhyroid dogs. } Am
Vel Med Assoc. 212:1572-1575.
10. Scoll-Moncrieff }CR, NeIson RW, runer }M and WiIIiams DA 1998 Comarison of serum
concenlralions of lhyroid-slimuIaling hormone in heaIlhy dogs, hyolhyroid dogs, and
eulhyroid dogs vilh concurrenl disease } Am Vel Med Assoc 212: 387-391.
11. Mei| I, MoI }A and Ri|nberk A 1996 Thyroid-slimuIaling hormone resonses afler singIe
adminislralion of lhyrolroin-reIeasing hormone and combined adminislralion of four
hyolhaIamic reIeasing hormones in beagIe dogs. Domesl Anim IndocrinoI 13: 465-468.
12. Mei| I, MoI }A, evers MM and Ri|nberk A 1997 ResiduaI iluilary funclion afler
lransshenoidaI hyohyseclomy in dogs vilh iluilary-deendenl hyeradrenocorlicism }
Indocr 155: 531-539.
13. Larsen IR, Davis TI, SchIumberger M-}, Hay ID 2003 Thyroid hysioIogy and diagnoslic
evaIualion of alienls vilh lhyroid disorders. In: WiIIiams Texlbook of IndocrinoIogy, 10
lh

ed. Larsen IR, Kronenberg HM, MeImed S, IoIonsky KS (Ids.) IhiIadeIhia, IA: W
Saunders. : 331-373.

Chaler 10
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14. Scoll-Moncrieff }CR and NeIson RW 1998 Changes in serum lhyroid-slimuIaling hormone
concenlralion in resonse lo adminislralion of lhyrolroin-reIeasing hormone lo heaIlhy
dogs, hyolhyroid dogs, and dogs vilh concurrenl disease. } Am Vel Med Assoc 213: 1435-
1438.
15. Shairo LI and Surks MI 2001 Hyolhyroidism. In: IrinciIes and Iraclice of IndocrinoIogy
and MelaboIism. 3
rd
ed. ecker KL (ed). IhiIadeIhia: Liincoll WiIIiams & WiIkins.: 445-
454.
16. SarIis N}, ruckner-Davis I, Doman }L, SkaruIis MC 1997 MRI-demonslrabIe regression of
a iluilary mass in a case of rimary hyolhyroidsm afler a veek of acule lhyroid hormone
lheray. } CIin IndocrinoI Melab 82: 808-811.
17. Sucuira MS, Camargo II, NickoIoff IL, AIderson IO, Wagner HN }r 1983 The roIe of
99m
Tc
erlechnelale ulake in lhe evaIualion of lhyroid funclion. Inl } NucI Med ioI 10: 29-33.
18. Na AMI, IoIIak YWIA, van den rom WI, Ri|nberk A 1994 Quanlilalive asecls of lhyroid
scinligrahy vilh erlechnelale (
99m
TcO4
-
) in cals. } Vel Inlern Med 8: 302-303.
19. Adams WH, DanieI G, Ielersen MG, Young K 1997 Quanlilalive
99m
Tc-erlechnelale lhyroid
scinligrahy in normaI beagIes. Vel RadioI UIlrasound 38: 323-328.
20. IeIdman and NeIson 1996. Hyolhyroidism. In: Canine and IeIine IndocrinoIogy and
Reroduclion. 2
nd
edilion. IeIdman IC and NeIson (Ids.). W Saunders Comany
IhiIadeIhia. I: 55-85.
21. Ri|nberk A 1996 Thyroids. In: CIinicaI IndocrinoIogy of Dogs and Cals. Ri|nberk A (ed).
Dordrechl, KIuver Academic IubIishers, : 35-39.
22. Iaggiano M, CriscuoIo T, Graziani M, Iorio S, Iisano G, Sinisi AA, Sinisi AM and eIIasleIIa A
1985 Iersilenl TRH-induced grovlh hormone reIease afler shorl-lerm and Iong-lerm L-
lhyroxine reIacemenl lheray in rimary congenilaI hyolhyroidis. CIin IndocrinoI 23:61-
66.
23. VareIa C, Cacicedo L, Iernandez G, de Ios IraiIes T and Sanchez Iranco I 1991 InfIuence of
hyolhyroidism duralion on deveIomenlaI changes in lhe hyolhaIamic faclors imIicaled
in grovlh hormone secrelion in lhe maIe ral. NeuroendocrinoIogy 54: 340-345.
24. Coiro V, VoIi R, Carelli L, Seroni G, Marchesi C, Vescovi II, Caffarri G, CoIIa R, Rossi G,
DavoIi C and Chiodera I 1994 InfIuence of lhyroid slalus on lhe aradoxicaI grovlh hormone
resonse lo lhyrolroin-reIeasing hormone in human obesily. MelaboIism 43: 514-517.
25. Hanev K, Ulsumi A, Sugavara A, Tanaka A, Iukazava H, Yoshida K and Abe K 1995
Inhanced Iasma GH resonses lo simuIlaneous adminislralion of TRH and GHRH in
alienls vilh rimary hyolhyroidism. IndocrinoI } 42: 43-47.
26. Rulleman GR, SloI R, Ri|nberk A, LoeffIer S, akker }A, evers MM, MeuIenberg IM and
Iigenmann }I 1987 Medroxy-rogeslerone acelale adminislralion lo ovariohyslereclomized,
oeslradioI-rimed beagIe bilches. Iffecl on secrelion of grovlh hormone, roIaclin and
corlisoI. Acla IndocrinoI 114: 275-282.


Samenvallende besreking en concIusies
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193

27. VidaI S, Horvalh I, Kovacs K, Cohen SM, LIoyd RV, Scheilhauer W 2000.
Transdifferenlialion of somalolrohs lo lhyrolrohs in lhe iluilary of alienls vilh
rolracled rimary hyolhyroidism. Virchovs Arch 436: 43-51
28. VaIenla L}, Tamkin }, Soslrin R, IIias AN, Iisenberg H 1983 Regression of a iluilary
adenoma foIIoving Ievolhyroxine lheray of rimary hyolhyroidism. IerliI SleriI 40: 389-
392.
29. Dam|anovic S, Ioovic V, Ielakov M, D|urovic M, Dieguez C and Casanueva II 1996
Iiluilary enIargemenl due lo rimary hyolhyroidism: grovlh hormone resonse lo GHRH,
GHRI-6 and GHRH Ius GHRI-6. } Iedialr IndocrinoI Melab 9: 549-553.
30. Khava|a NM, Taher M, arham MI, Naser AA, Hadidy AM, Ahmad AT, Hamamy HA,
Yaghi NA, and A|Iouni KM. 2006 Iiluilary enIargemenl in alienls vilh hyolhyroidism
Indocr Iracl 12: 29-34.
31. ZeIissen IM}, Croughs R}M, Koerschaar HII, de ruin TW, Hendriks M}, van'l VerIaal
}W 1989 Irimary hyolhyroidism mimicking iluilary adenoma. Nelh } Med 35: 95-99.
32. Horvarlh I, LIoyd RV, Kovacs K 1990 IroyIlhiouracyI-induced hyolhyroidism resuIls in
reversibIe lransdifferenlialion for somalolrohs inlo lhyroideclomy ceIIs. A morhoIogic
sludy of lhe ral iluilary incIuding immunoeIeclron microscoy. Lab Invesl 63: 511-520.
33. ViIIaIobos C, Nuez L, Garcia-Sancho } 2004 Anlerior iluilary lhyrolroes are
muIlifunclionaI ceIIs Am } IhysioI IndocrinoI Melab 287: I1166-70.
34. VidaI S, Horvalh I, Kovacs K, LIoyd RV, Smylh HS. ReversibIe lransdifferenlialion:
Inlerconversion of somalolrohs and Iaclolrohs in iluilary hyerIasia. Mod IalhoI 2001,
14:20-28.
35. Corlese L, OIiva G, Verslegen }, CiarameIIa I, Iersechino A 1997 HyerroIaclinemia and
gaIaclorrhea associaled vilh rimary hyolhyroidism in a bilch. } SmaII Anim Iracl 38: 572-
575.
36. Raber W, GessI A, Novolny I, Vierhaer H 2003 HyerroIaclinaemia in hyolhyroidism:
cIinicaI significance and imacl of TSH normaIizalion. CIin IndocrinoI 58: 185-191.
37. Kooislra HD, Okkens AC 2002 Secrelion of grovlh hormone and roIaclin during
rogression of lhe IuleaI hase in heaIlhy dogs: a reviev. MoI CeII IndocrinoI 197: 167-172.
38. Okkens AC, DieIeman S}, Kooislra HS, evers MM 1997 IIasma concenlralions of roIaclin in
overlIy seudoregnanl Afghan hounds and lhe effecl of melergoIine. } Rerod IerliI, SuI
51: 295-301.
39. GaIac S, Kooislra HS, ulinar }, evers MM, DieIeman S}, Voorhoul G, Okkens AC 2000
Terminalion of midgeslalion regnancy in bilches vilh agIerislone, a rogeslerone recelor
anlagonisl. TheriogenoIogy 53: 941-950.





Chaler 10
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194












































Dankwnnrd

Curricu!um Vitac

Pub!icatinns

List nI abbrcviatinns

Dankvoord
_________________________________________________________________________________

196
Dankvoord
_________________________________________________________________________________

197

Dankwnnrd

Dil roefschrifl is hel resuIlaal van hel verk van veeI mensen, die hier in de Ioo
van deze |aren o een direcle of een indirecle manier bi| belrokken zi|n geveesl.
Ondanks een soms eenzaam gevoeI, is hel een feil dal ik dil verk nooil aIIeen had
kunnen doen.
Irof. Dr. Ad Ri|nberk. Woorden zi|n onvoIdoende om u le danken voor zo veeI
geduId en vaardevoIIe li|d li|dens hel begeIeiden van dil roefschrifl. Uv enorme
inzel, uv grondige kennis, uv assie voor de endocrinoIogie en voor de inlerne
geneeskunde, en uv vi|ze en obouvende kriliek zi|n bevonderensvaardig.
U heefl me Ialen zien dal hard verken, en zeker een onmisbare vaslberadenheid,
nodig zi|n om een doeI le bereiken.
Leefli|d is een slaal van de zieI en geesl. AIs men zi|n nieuvsgierigheid,
enlhousiasme en innerIi|k energie kan behouden, uilslraIen en deIen, dan heefl
men hel geheim van de eeuvige |eugd.
In hel bi|zonder viI ik ook uv vrouv, mevrouv CarIa Ri|nberk, heeI harleIi|k
bedanken voor haar gaslvri|heid en de geIegenheid die zi| bood om bi| uv lhuis le
kunnen verken.
Dr. Hans Kooislra. Ondanks een heeI drukke agenda en meerdere ro|eclen die
legeIi|k Iieen en nog sleeds Ioen, kon ik |ouv absoIuul onmisbare huI kri|gen
mel mi|n roefschrifl. }ouv uilgebreide en diee kennis van de endocrinoIogie en
de inlerne geneeskunde is onvoorsleIbaar. }ouv osilieve insleIIing en
enlhousiasme zi|n loch besmelleIi|k en gaven me dal exlra duvl|e dal nodig vas
om voI le houden en door le gaan.
Dr. }an MoI. }ouv uilgebreide ervaring mel onderzoek heefl mee gehoIen om dil
roefschrifl lol sland le brengen en le onlvikkeIen.
Dr. Mari|ke Ieelers. }ouv erfeclionislische en recieze manier van verken is voor
|ouv coIIega's, de eigenaren en de alinlen een enorm voordeeI geveesl. Geen
ander onderzoek over hyolhyreodie bi| de hond heefl de mogeIi|kheid gehad om
de funclie en de morfoIogie van de schiIdkIier le onderzoeken in zo'n grool aanlaI
alinlen. Mel een gerusl gevoeI Iielen ve de schiIdkIier biolen van onze
alinlen nemen, mel de zekerheid dal hel voImaakl zou verIoen.
Irof. Dr. }aa van Di|k. Ti|dens de bi|zondere momenlen van hel beki|ken van de
alhoIogie rearalen van de schiIdkIieren, heb ik mel echle inleresse geIuislerd
naar uv heIdere beschri|ving en uilIeg over de alhoIogische bevindingen.
Overluigend hel resuIlaal van veIe |aren rofessioneIe ervaring en kennis. Uv
beIangsleIIing en nieuvsgierigheid voor hel onderver varen heeI omerkeIi|k.
Uv anekdolen over de inleressanle gesrekken lussen alhoIogen en cIinici van
|aren geIeden over de alinlen, hun ziekleverschi|nseIen, en alhoIogische
bevindingen, varen fascinerend.
Dankvoord
_________________________________________________________________________________

198

Dr. Ted van den Ingh. Uv huI bi| hel alhoIogisch onderzoek en de
immunohislochemie van de hyofyses zi|n een cruciaIe bi|drage geveesl voor dil
onderzoek.
Dr. RoseIinda van de VIugl-Mei|er. AIIe CT-scans van de hyofyses zi|n o een
rofessioneIe en belrouvbare manier door |ou uilgevoerd. }ouv inzel en
enlhousiasme zi|n een onmisbare conlribulie geveesl om een deeI van de
resuIlalen van dil roefschrifl, vaar vi| zo lrols o zi|n, le kri|gen.
Yvonne IoIIak. }ouv essenliIe bi|drage aan dil ro|ecl is giganlisch. Vanaf de
eersle dagen heb |e builengevoon hard geverkl om aIIe scinligrafien uil le
voeren. Inlensief, li|drovend, erfecl verk. }ouv kennis, deskundigheid en
ervaring zi|n uilzonderIi|k. IindeIoos heb |e de grafieken, en figuren gemaakl
zonder le Ialen merken dal hel le veeI verd. }ouv rofessionaIileil en
erfeclionislische aard zi|n uniek en hel is voor mi| een enorm Iezier geveesl mel
|ou le hebben samengeverkl, een echle Iuxe.
Harry van IngeIen. Onderzoek zonder Iogisliek is onmogeIi|k. }ouv vermogen om
aIIes le regeIen en cordineren, en in een razende lemo en vIekkeIoos le Ialen
verIoen, is een van |ouv zeer osilieve eigenschaen. Mel bevondering kan ik
nog sleeds voor mi| zien, hoe |e kalhelers Iaalsle, bIoedmonslers afnam, en
afhandeIde mel een sneIheid en nauvkeurigheid die onmogeIi|k le verbeleren is.
}ouv onvermoeibare en rofessioneIe huI en medeverking hadden nooil gemisl
kunnen vorden.
Mi|n coIIega's dierverzorgers en dierverIegers van hel Dearlemenl Geneeskunde
van GezeIschasdieren. AI die |aren varen |uIIie een enorme sleun en huI in hel
uilvoeren van de funclieleslen en hel beaIen van de hormoonrofieI rolocoIIen.
Mevrouv Monique van WoIferen, Mevrouv }eannelle WoIfsvinkeI en de andere
medeverkers van de diverse Iaboraloria. Zonder |uIIie giganlische samenverking
en |uIIie Iangdurige verk om aIIe beaIingen uil le voeren, vas dil roefschrifl
onmogeIi|k geveesl.
Mi|n coIIega's van de afdeIing aneslhesioIogie. De CT-scans van de hyofyses, en
de schiIdkIier chirurgie moeslen naluurIi|k zonder een enkeI incidenl gebeuren.
Ien enorme veranlvoordeIi|kheid. }uIIie velen hel aI: zonder |uIIie vas hel niel
mogeIi|k geveesl.
Mi|n aranimfen: Aslrid van Dongen en Chrisline Iiek. Mel veeI Iezier ki|k ik
lerug naar zo veeI |aren van samenverking o de KIiniek. Vanaf hel begin aI
varen |uIIie aIli|d kIaar om le heIen en le ondersleunen. Hel is een voorrechl mel
zuIke rofessioneIe en echle fi|ne coIIega's le mogen verken.
In de veIe |aren o de KIiniek voor GezeIschasdieren heb ik de kans gehad om
verschiIIende coIIega's le onlmoelen (sommige hebben inmiddeIs de KIiniek aI
verIalen) mel vie ik gesrekken over ambilies, carriere en rive gedeeId heb.
i|zondere en unieke ervaringen die soms een diee indruk gemaakl hebben.
Dankvoord
_________________________________________________________________________________

199

CoIIega's die ieder o hun eigen manier hebben gehoIen mel hel afronden van de
romolie: ruiIingen van oIi's en dienslen, overname van verk- en hoor-
coIIeges, huI mel de Iayouls voor hel boek, enzovoorl.
Mi|n Saanse coIIega's DoIores Ierez (HosilaI CIinico Velerinario, IacuIlad de
Velerinaria, Universidad ComIulense Madrid) en CarIos MeIian (HosilaI CIinico
Velerinario en CIinica Velerinaria AlIanlico, Las IaImas de Gran Canaria). In de
Iaalsle |aren heb ik de kans gehad om mel |uIIie samen le verken. In hel bi|zonder
mel hel boek: ManuaI de IndocrinoIogia de Iequeos AnimaIes (2008). }uIIie
doorzellingsvermogen en inzel zi|n een insiralie. Mel LoIa (DoIores Ierez) heb ik
hel geIuk om een bi|zondere vriendscha le hebben die lerug gaal naar onze
sludenlenli|d in Madrid. (In Ios uIlimos aos he lenido Ia osibiIidad de coIaborar
con vosolros con eI Iibro: ManuaI de IndocrinoIogia de Iequeos AnimaIes (2008).
Vueslra caacidad de lraba|o y sueracin son una insiracin. Con LoIa lengo
ademas Ia suerle de lener una amislad eseciaI que se remonla a nueslros liemos
de esludianles en Madrid).
De Ieiding van hel Dearlemenl Geneeskunde van GezeIschasdieren, viI ik
danken voor de kans om dil roefschrifl lol sland le kunnen brengen en af le
ronden.
}aren lerug kreeg ik de geIegenheid om de oIeiding lol seciaIisl in Ulrechl le
voIgen. Deze uilzonderIi|ke kans heb ik le danken aan de inzel en invIoed die
beaaIde mensen loen voor mi| uilgeoefend hebben. Ik ben me aIli|d heeI bevusl
geveesl dal zonder hun sleun desli|ds hel me nooil geIukl zou zi|n. Ik ben en zaI er
aIli|d onlzellend dankbaar voor zi|n.
Mi|n ouders. Door |uIIie insanning en onvoorvaardeIi|ke sleun is mi|n droom om
dierenarls le vorden verkeIi|kheid gevorden. (Mis adres. Vueslro sacrificios y
ayuda incondicionaI han hecho osibIe mi sueo de ser velerinaria).
Iran, Nuria en MigueI. }uIIie energie, nieuvsgierigheid, en enlhousiasme eisl een
lo condilie om |uIIie rilme bi| le houden. }uIIie geven me iedere dag insiralie,
krachl en een comIeel geIukkig gevoeI.
Andre, de man van mi|n harl en mi|n Ieven. }ouv IiefdevoIIe ondersleuning en
begri is onbeschri|feIi|k. Samen hebben ve de mooisle beIevenissen en ro|eclen
in onze Ievens meegemaakl en hoeIi|k mogen ve in de loekomsl nog veIe
meemaken. Aan |ou en de kinderen is dil roefschrifl ogedragen.
CurricuIum Vilae
_________________________________________________________________________________

200

Curricu!um vitac

The aulhor of lhis lhesis vas born in II IerroI (La Corua, Sain) on lhe 27
lh
}uIy
1966. Afler a rimary schooI educalion in II IerroI (Sain), and secondary schooI
educalion al lhe High schooI Salurnino Monlo|o (II IerroI), and al lhe High schooI
Irincie IeIie (Madrid), she slarled velerinary medicine al lhe IacuIly of
Velerinary Medicine al ComIulense Universily, Madrid (Sain) in 1984. She
gradualed in 1989 and vorked as an assislanl leacher and cIinician al lhe IacuIly of
Velerinary Medicine al ComIulense Universily, Madrid (Sain). In 1994 she
slarled her inlernshi/residency lraining rogram al lhe Dearlmenl of CIinicaI
Sciences of Comanion AnimaIs, IacuIly of Velerinary Medicine, Ulrechl
Universily, (The NelherIands). In 1999 she successfuIIy assed lhe board
examinalion of lhe Iuroean CoIIege of Velerinary Medicine of Comanion
AnimaIs (ICVIM-CA) and became a DiIomale ICVIM-CA. Al lhe Dearlmenl of
CIinicaI Sciences of Comanion AnimaIs, IacuIly of Velerinary Medicine, Ulrechl
Universily, she has erformed her IhD and has been vorking lo dale as an
Inlernisl.


Lisl of ubIicalions
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201

List nI pub!icatinns

1. IscoIar I, Ierez D, Diaz-Isieira MM, Rodriguez A 1993 Canine Ianconi
Syndrome. A case reorl. }ournaI of SmaII AnimaI Iraclice 34: 567-570.
2. oersma A, ZonnevyIIe }, Sanchez MA, Diaz-Isieira MM 1995
Irogressive alaxia in a rollveiIer dog. Velerinary QuarlerIy 17:108-109.
3. Diaz-Isieira MM, IscoIar I, eIIanalo }, Rodriguez M 1995 Slruclure
and comosilion of equine uroIilhs. Iquine Velerinary Science 15:27-34.
4. Diaz-Isieira MM, IscoIar I, eIIanalo }, Medina }A 1995 CryslaIIine
comosilion of equine urinary sabuIous deosils. Scanning Microscoy
9:1071-1079.
5. Diaz-Isieira MM, IscoIar I, eIIanalo }, Rodriguez M 1996 Minor
consliluenls of sabuIous maleriaI in equine urine. Research in Velerinary
Science 60:238-242.
6. Diaz-Isieira MM, and SIaendeI R} 1997 A case of aulochlhonous
canine Ieishmaniasis in lhe NelherIands. Velerinary QuarlerIy 19:69-71.
7. Diaz-Isieira MM, Viehoff I, and NickeI RI 1998 Idioalhic delrusor-
urelhraI dyssynergia in dogs: a relroseclive anaIysis of 22 cases. }ournaI
of SmaII AnimaI Iraclice 39:264-270.
8. Diaz-Isieira MM, Vink-Nooleboom M, van den Ingh TSGAM,
Rolhuizen } 1999 Thrombosis of lhe orlaI vein in a minialure schnauzer.
}ournaI of SmaII AnimaI Iraclice 40:540-543.
9. Kooislra HS, Diaz-Isieira MM, MoI }A, van den rom WI, Ri|nberk A
2000 Secrelion allern of lhyroid-slimuIaling hormone in dogs during
eulhyroidism and hyolhyroidism. Domeslic AnimaI IndocrinoIogy 18:19-
29.
10. Lee WM, Diaz-Isieira MM, MoI }A, Ri|nberk A, Kooislra HS 2001
Irimary hyolhyroidism in dogs is associaled vilh eIevaled GH reIease.
}ournaI of IndocrinoIogy 168:59-66.
11. Diaz-Isieira MM, MoI }A, Ieelers MI, IoIIak YWIA, Iversen L, van
Di|k }I, Ri|nberk A, Kooislra HS 2007 Assessmenl of lhyroid funclion in
dogs vilh Iov Iasma lhyroxine concenlralion. }ournaI of Velerinary
InlernaI Medicine 21:25-32.
12. Diaz-Isieira MM, GaIac S, MoI }A, Ri|nberk A, Kooislra HS 2007
Thyrolroin-reIeasing hormone-induced grovlh hormone secrelion in
dogs vilh rimary hyolhyroidism. Domeslic AnimaI IndocrinoIogy
34:176-181.
13. Diaz-Isieira MM, MoI }A, van den Ingh TSGAM, van de VIugl-Mei|er
RH, Ri|nberk A, Kooislra HS 2008 IunclionaI and morhoIogicaI changes
in lhe adenohyohysis of dogs vilh rimary hyolhyroidism: Loss of
Lisl of ubIicalions
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202

TSH hyersecrelion, hyersomalolroism, hyoroIaclinemia, and
iluilary enIargemenl vilh lransdifferenlialion. Domeslic AnimaI
IndocrinoIogy 35:98-111.
14. Diaz-Isieira MM, MoI }A, Ri|nberk A, Kooislra HS AdenohyohyseaI
funclion in dogs vilh rimary and in non-lhyroidaI iIIness. Acceled for
ubIicalion in }ournaI of Velerinary InlernaI Medicine.
15. Diaz-Isieira MM 2008 GIanduIa liroidea. Hioliroidismo canino. In:
ManuaI de IndocrinoIogia de Iequeos AnimaIes. C. MeIian y M.D.
Ierez-AIenza (Ids.). Irimera edicin. MuIlimedica Idiciones Velerinarias,
arceIona, Isaa. I: 85-107.



Lisl of abbrevialions
_________________________________________________________________________________

203

List nI abbrcviatinns.


-MSH meIanocyle slimuIaling hormone
ACTH adrenocorlicolroic hormone
ANOVA anaIysis of variance
AT4 anlibodies againsl T4
AT3 anlibodies againsl T3
ATI adenosine lrihoshale
AUC area under lhe curve
cAMI cycIic adenosine monohoshale
CRH corlicolroin-reIeasing hormone
CT comuled lomograhy
DIT diiodolyrosine
IDTA elhyIene diamine lelraacelic acid
ISH foIIicIe-slimuIaling hormone
GH grovlh hormone
GHRH grovlh hormone-reIeasing hormone
GnRH gonadolroin-reIeasing hormone
IGI-I insuIin-Iike grovlh faclor-I
IU inlernalionaI unils
IV inlravenous
LH Iuleinizing hormone
MIT monoiodolyrosine
MRI magnelic resonance imaging
mRNA messenger ribose nucIeic acid
NIS sodium-iodide symorler
NTI non lhyroidaI iIIness
IH rimary hyolhyroidism
IRL roIaclin
rT3 reverse 3,5,3',-L-lriiodolhyronine
SD slandard devialion
TgAbs lhyrogIobuIin anlibodies
TIO lhyroid eroxidasa
TRH lhyrolroin reIeasing hormone
TSH lhyroid slimuIaling hormone or lhyrolroin
T4 3,5,3',5',-L-lelraiodolhyronine
T3 3,5,3',-L-lriiodolhyronine