BIOLOGY

OF

REPRODUCTION

35, 84-91

(1986)

Prolactin Granulosa

Modulates Steroidogenesis by Rat Cells: I. Effects on Progesterone1

and S. E. VINCENT of Biological of Arts and Sciences Sciences

J. E. FORTUNE2 Section of Physiology Women s Studies Program

Department of Physiology

in the Division in the College Cornell University

and

in the College of Veterinary York 14853

Medicine

Ithaca, New

ABSTRACT
Either

cellsfrom

testosterone or follicle-stimulating hormone (FSH) stimulates progesterone secretion by granulosa rats but the combination of the two hormones increases progesterone production in a synergistic

manner. We have investigated the effects of graded doses of prolactin (0, 0.02, 0.2, 2, or 10 pg/mi) alone or in combination with testosterone (0.5 pM), FSH (300 ng/mi), or FSH + testosterone on progesterone secretion by granulosa cells at two stages of differentiation. Relatively undifferentiated granulosa cells from immature, dietbystilbestrol-treated, hypopbysectomized (HPX) rats were cultured in defined (serum-free) medium for 3 days. More highly

follicles

of 30-day-old

differentiated granulosa cells were obtained rats induced to undergo an estrous were cultured in medium containing

on the morning of proestrus from the preovulatory cycle by injection with 4 IU pregnant mares serum 10% fetal bovine serum.

gonadotropin;

these cells

Prolactin

alone

did not

enhance the negligible secretion of progesterone cells from proestrous rats. Prolactin significantly on cells from with prolactin, both HPX and proestrous secretion was significantly the direction progesterone by cells from

by cells from HPX rats, enhanced the stimulatory cultures containing proestrous rats was

but increased progesterone effects of testosterone both significantly HPX from FSH
+

secretion by or FSH alone were treated secretion with both prolactin

rats. When

testosterone whereas

enhanced,

by cells from HPX rats FSH and testosterone,

+

depressed. Therefore when cells from of the effect of prolactin was reversed that whether

rats were cultured that observed with cultured was due

FSH or testosterone alone, and from #{247} FSH + testosterone. To determine

observed
the

when difference

cells from proestrous rats were in results for the two models different 10% culture systems, serum, With these with FSH,

with prolactin to differences were reprolac+

in the stage of differentiation of the granulosa cells peated with cells from HPX rats and with medium
tin stimulated tosterone. In

or to the containing alone

the experiments culture conditions testosterone,

progesterone
summary,

secretion prolactin in granulosa however, medium.

when

used

or in combination

or FSH

tes-

general

stimulated

secretion

of progesterone

by both

undifferentiated

and

relatively differentiated and FSH + testosterone; moving serum from the

cells from rats its enhancement

and enhanced of the effect

the stimulatory effects of FSH #{247} testosterone

of FSH, testosterone, could be reversed by re-

INTRODUCTION

In mammalian species prolactin is best known for its action in regulating mammary gland function and as an essential part of the luteotropic complex in rodents may opment conditions (see review by Smith, also play a role in the since certain in which ovulation 1980). control However, prolactin of follicular develand is suppressed pathological (e.g., lac-

tational mia) are prolactin evidence

anestrus/amenorrhea accompanied (see that review prolactin

by by

high

and hyperprolactinelevels of circulating 1979). There follicular is de-

McNeilly, may suppress

velopment indirectly by affecting of gonadotropin secretion (see 1979, time, isolated 1980; prolactin ovarian of Prolactins ovulatory actions on Tresguerres et may directly cells (see

the rate and pattern reviews by McNeilly, same by et al.,

physiological

al., 1981). At the effect steroidogenesis by McNeilly

review

1982).
ment
Accepted December 20, 1985. Received February 14, 1983. This study was supported by NIH 2 Reprint requests: J. E. Fortune, Cornell University, Ithaca, NY 14853.

action in suppressing follicles, may thus both gonadotropin

the developbe complex, secretion on been progesterreported. and

involving
grant 823 HD-14584. Veterinary

Research

Tower,

ovarian steroidogenesis. A variety of effects of prolactin one production by ovarian cells has 84

PROLACTIN

MODULATES

PROGESTERONE

SECRETION

85 of oocytes on et al., 1976).

McNatty tions

and of prolactin

coworkers inhibited

found

that

high

concentra-

production

of progesterone

30, the

and ovulation morning of

of a normal number Day 31 (Armstrong

by developing follicles from mice (McNatty et al., 1976) and by granulosa cells obtained from women at various stages of the cycle (McNatty et al., 1974). However, low concentrations of prolactin seemed necessary to maintain of human granulosa lactin cells secretion 1980). reported In progesterone secretion cells (McNatty et al., secretion follicles, follicles by but in cultures 1974). Progranulosa stimulated et a!., (1980) syner-

Animals were killed by cervical dislocation on the morning of Day 30, the day of proestrus. This model is believed to approximate closely the events of the adult granulosa differentiated Isolation Ovaries placed (MEM; and estrous cells cycle than Culture (Armstrong those from et al., HPX rats. Cells aseptic conditions, 1976) would and the obtained at proestrus be more

inhibited progesterone from small porcine by cells from large addition, Veldhuis that estradiol and

of Granulosa under

(Veldhuis

were in modified Dorrington

removed

and Hammond prolactin acted

Eagles minimum et al., 1975)

essential medium and removed from The with a into the the cells and

gistically on granulosa cells from small porcine follicles both to inhibit secretion of progesterone early in culture and to stimulate its secretion later in culture. Therefore, the effects of prolactin on granulosa cells seem dependent on the dose of prolactin, the presence of other stage of differentiation further investigated of progesterone cells maturation granulosa follicular hormones in the medium, and the of the granulosa cells. We have prolactins effects on the secretion by granulosa at with two graded cells by doses culturing of prolactin and/or rat different stages of

their bursae under a dissecting microscope. ovaries of HPX rats were pierced repeatedly fine insect pin to release granulosa cells medium. The ovaries were discarded, and were collected a by centrifugation, resuspended, Proestrous from the were cut scraping Cells counted in (10-12/rat) PMSG-treated the granulosa layer with animals were and counted a hemacytometer. were dissected rats. The follicles cells fine

follicles ovaries of in half and the from thecal several

obtained combination hormone

isolated by glass needle.

alone or in follicle-stimulating

with testosterone (FSH).

pooled for each experiment, as described above.

resuspended alone or in (0.5 pM), prolactin Testosterone to achieve ml medium). the

MATERIALS Animals Immature chased from Granulosa obtained rats that female Blue

AND METHODS

Cells were cultured in modified MEM modified MEM containing testosterone FSH (NIH-FSH-S8, 300 ng/ml), and/or (NIH-P-S dissolved desired The did 12; 0.02, 0.2, was (100 of ethanol testosterone. 2 or added 10 pg/ml). to MEM was in ethanol concentration same amount not contain

Sprague-Dawley Spruce Farms

rats were (Altamont,

purNY).

p1 ethanol/30

cells used in these experiments were from two different experimental models: had been hypophysectomized (HPX) and

added to media that In some experiments

treated with diethylstilbestrol (DES), and rats treated with pregnant mares serum gonadotropin (PMSG). Hypophysectomies were performed by the supplier at 21 days of age. Rats were injected s.c. with 2 mg DES in 0.1 ml sesame oil on Days 24-28 and killed by cervical dislocation on Day 29. Since DES is a mitogen for granulosa cells, ovaries of these rats contained large numbers of granulosa cells that had not been stimulated by were relatively intact immature endogenous undifferentiated. rats were gonadotropins and thus In the second model, injected s.c. with 4 IU

with cells from HPX rats and in all experiments with cells from proestrous rats, media contained 10% fetal bovine serum (GIBCO, Grand Island, NY). Treatments were applied to replicate cultures within each experiment two Cells dishes and groups each were (Costar, experiment cultured in Cambridge, was repeated with at least of rats. plastic, 24-well multiwell MA). Cultures were in 50 p1 MEM to were maintained gassed with 50% were removed were frozen and for

initiated by adding 200,000 cells 450 p1 treatmentmedium. Cultures at 37#{176}C in a humidified incubator C02, 95% air for 3 days. replaced daily. Medium Media samples

PMSG (Ayerst Laboratories, Inc, New York, NY) on Day 28 to initiate an estrous cycle. This treatment causes follicular growth, estrogen secretion, luteinizing hormone (LH) on the afternoon a surge of of Day

subsequent radioimmunoassay. observed daily with an inverted the general attachment appearance and flattening of the

Cultured microscope of the Prolactin

cells were to monitor cells and appeared the

cultures.

86 to have no effect of the on cells and cell viability or on

FORTUNE

AND

VINCENT

the

general

appearance

in culture.

A.HPX
Radioimmunoassays Statistics Aliquots of culture medium were measured for progesterone without extraction by radioimmunoassay, as described previously (Fortune and Eppig, 1979). The progesterone antiserum, prepared at Cornell University
by

6O Ui (I)

50

J.

E.

Hixon, with

cross-reacts

11.6%

with Scs-dihydroprogesterone, gestins tested and <0.1% trogens expressed analysis tested per of (Fortune 100,000 variance, were were and Accumulation

<10% with other prothe androgens and es1978).

o o o o

w 40

Armstrong, in the Data were individual

of progesterone cells. and

medium subjected comparisons

was to

20
C

between means range test. Data heterogeneity

made using transformed was RESULTS

Duncans multiple to logarithms when I.

Ui

10
...........

.........

of variance

present.

.,2zz::::::-.s:.
50

B.
Effects of Prolactin on Granulosa (I) 40
LU

Proestrous
--

Cells Obtained and Cultured

from HPX Rats in Defined Medium

I,

The effects of prolactin on secretion of progesterone by granulosa cells from HPX rats were assessed by culturing cells with prolactin alone and in combination were known tion with testosterone and/or FSH. These and progesterone and treatments FSH are secreselected because testosterone to interact in stimulating by granulosa cells (Armstrong

30
LU

/ -F

20
LII
.
/ lt I,J#{149}

,
/ / ,

--.--

(9
,_

,.-,

Dorring-

ton, 1976; Nimrod and Lindner, 1976). Both FSH and testosterone alone slightly, but significantly (p<0.001), stimulated secretion of progesterone, but the combination of FSH and testosterone secretion of progesterone in an apparently manner (p<O.OOl, Fig. 1A). These results to those (1976) first and reported Nimrod by and cells absence Armstrong Lindner were of and (1976) increased synergistic are similar Dorrington using to prolactin testosterone), 2A). The secretion the

LU

e_,_-..x

-f
HPX (10%

30

Seru,,4T

HPX rat model. When granulosa alone (i.e., in the progesterone stimulatory

exposed FSH or

secretion effect of

was negligible (Fig. FSH on progesterone

DAYS

OF

CULTURE

(Fig. 1A) was enhanced in a dose-dependent fashion by adding prolactin to the culture medium (Fig. 2B; p<0.01); 0.2 g prolactin/ml produced a maximal effect, except on the third day of culture. Although cultures production containing of progesterone alone, was low in testosterone or testosterone

FIG. 1. Cumulative secretion of progesterone by granulosa cells (ngl 100,000 cells SEM) over 3 days of culture in medium alone (C), with FSH (F; 300 ng/mI), testosterone (T; 0.5 MM), or FSFI + testosterone (T#{247}F). A) Cells obtained from hypophysectomized (HPX) rats and cultured in defined medium (n = 6 cultures). B) Cells from proestrous follicles cultured in medium containing 10% serum (n = 12 cultures). C) Cells from HPX rats, cultured in medium containing 10% serum (n = 8 cultures).

PROLACTIN

MODULATES

PROGESTERONE

SECRETION

87 effect HPX rats (p<0.001). (Fig. 1A), As with combina-

H PX
*A
LU 0.5 C)
+1
U)
-

tenfold) granulosa tion of secretion (p<0.001). In the of

stimulatory cells from

the

Pri

FSH and testosterone increased progesterone even more, in a greater than additive fashion with prolactin of prolactin in alone increased to (Fig. 3A), secretion the lack of

*B
1.5

PrI

FSH

cultures treated two highest doses (p<0.05)

progesterone

contrast

0
0

0
a,

F111
C.
PrI+T
0.5

effect on granulosa cells from HPX rats (Fig. 3A vs. Fig. 2A, note difference in scale of the figures). Prolactin also increased progesterone secretion by cells cultured with FSH with or FSH, testosterone prolactin (Figs. at doses 3B,C). In combination of 0.2 pg

z
0
ILU C-)
LU
(1,

D.

PrI+FSH+T
A.
I0
LU (I)
+1
U)

PROESTROUS Pri

LU

z
0
LU

IC/)
LU

4
B. Pri
+

CD

0
0 0 .02 .2 2 10 0 .02 .2 2

FSH

0 DAY I

DAY2

DAY3

0 0
0

10 r

PROLACTIN

(tg/mI)

0
a,
C

FIG. 2. Progesterone secretion (ng/100,000 cells SEM) on each day of culture by granulosa cells obtained from hypophysectomized rats (HPX) and cultured in defined medium containing A) graded doses of prolactin (Pr!) alone (n = 8 cultures), B) prolactin in combination with FSH (300 ng/ml; n = 8 cultures), C) prolactin in combination with testosterone (T, 0.05 MM; n = 8 cultures), or D) prolactin in combination with FSH and testosterone (n = 6 cultures). #{149}Notethe difference in scale of the Y axis of panels A-C compared with panel D.

20

C.

PrI+T

z
I-

10

0
LU

0
LU

I 1j
D.
PrI+FSH+T
x.-

a.

U, LU

II
.1

in combination tin, the highest of progesterone contrast, when were treated decreased by 10 g prolactin Effects Cells

with dose about cultures

the

three

lower

doses

of prolac-

z
0

of prolactin fourfold containing

increased secretion (p<0.01; Fig. 2C). In FSH + testosterone secretion of 2 or

cr
LU

20

x.

with prolactin, about twofold (p<0.01; Fig. on Granulosa Rats from

progesterone in the presence 2D).

IC,) LU C:,

I0

0 .02 .2 2 10
DAY 1

0 .02 .2 2
DAY2

10

0 .02 .2 2
DAY 3

10

of Prolactin from Proestrous cells

PROLACTIN

(hg/mI)

Granulosa

proestrous of

rats

secreted

small,

but measurable amounts absence of added hormones alone slightly increased (p<0.001), while FSH had

progesterone in the (Fig. 1B). Testosterone progesterone production a much greater (sevento

FIG. 3. Progesterone secretion (ng/100,000 cells SEM) on each day of culture by granulosa cells obtained from proestrous rats and cultured in medium containing 10% serum and A) graded doses of prolactin (Pri) alone (n = 6 cultures), B) prolactin in combination with FSH (300 ng/ml; n = 6 cultures); C) prolactin in combination with testosterone (T, 0.05 MM; n = 12 cultures) or D) prolactin in combination with FSH and testosterone (n = 12 cultures).

88 or third did tion alone with Fig. FSH all days 3C). and greater days increased of culture the of had added progesterone (p<0.05) decline culture dramatic to and on the

FORTUNE second and but

AND

VINCENT

HPX *A Pri

(10%

serum)

ameliorated,

not prevent, over 3 days (Fig. 3B). testosterone of culture When testosterone,

in progesterone that occurred prolactin stimulatory of 2 or 10 pg/mI cultures had containing

producwith FSH effects (p<0.01; both on the pg/mi or secreand 0.5

0

In contrast, at doses

in combination on
LU

*B

Pri

FSH

U) +1
,,

1.5 1.0

prolactin

no effect

first day of culture; greater it prevented tion third Effects that days occurred of culture

but, at doses of 0.02 the decline in progesterone progressively (p<0.05; on Granulosa Fig. on 3D). the

second

0 0
C,

iI
*

C.

PrI#{247}T

of Prolactin

Cells Obtained from HPX Rats and Cultured with 10% Serum The results presented above show that when added to cultures containing testosterone or FSH, prolactin increased secretion of progesterone by granulosa cells from both HPX and proestrous rats. However, the effects of testosterone two terone under experimental prolactin and FSH on cultures were radically Prolactin cells stimulated granulosa but containing different inhibited from HPX secretion both in the progesrats by in the secre-

z
0
ILU

0
LU

40

C)
LU

0.

PrI+FSH+T

0
LU IU) LU CD

30

II

Iii

systems.

secretion by these conditions,

0
0

cells from modulatory

proestrous rats. effect of prolactin

This difference on progesterone

tion stimulated by the combination of FSH + testosterone might be due to the different stage of development of the granulosa cells from these two models or it could be due to the different, basic culture conditions provided (i.e., defined medium for HPX cells vs. defined estrous rats do defined repeat medium cells).
+

.02

.2

10

0.02.2210 DAY 2 (p.g/mI)

.02 .2 DAY 3

2 I0

DAY

I PROLACTIN

Since

10% fetal granulosa or flatten as Eagles on

bovine cells

serum for profrom proestrous in simple decided to rats cells with from

not attach media, such the experiments

effectively MEM, we from HPX

cells

FIG. 4. Progesterone secretion (ng/100,000 cells SEM) on each day of culture by granulosa cells obtained from proestrous rats and cultured in medium containing 10% serum and A) graded doses of prolactin (Pr!) alone (n = 8 cultures), B) prolactin in combination with FSH (300 ng/ml; n = 8 cultures), C) prolactin in combination with testosterone (T, 0.05 MM; n 8 cultures), or D) prolactin in combination with FSH and testosterone (n = 6 cultures). Note the difference in scale of the Y axis of panels A-C compared with panel D.

medium containing In the presence

10% serum. of 10% serum,

granulosa

HPX rats produced measurable quantities of progesterone, and secretion was increased by FSH alone and testosterone alone (p<0.001; Fig. 1C). Once again, the combination of these two hormones increased progesterone in a synergistic fashion (p<0.001). Treating lactin secretion culture (p<0.Ol; the alone in cells with the (0.02-10 pg/mi) various resulted doses of proin increased of the

third was

day

of culture. with the

When proiactin,

either

FSH progesterone

or testosterone secretion in (Fig.

combined

was increased response to 4B,C). lactin were culture greater

slightly 10 pg

but significantly (p<0.05) or the 2 and 10 g doses

of progesterone the presence Fig. 4A).

during the first two days of 2 or 10 g prolactin/mi was negligible on

In combination with FSH + testosterone, proincreased progesterone (Fig. 4D); its effects significant on the second and third day of (p<0.01). Doses of 0.2 pg proiactin/ml or produced a maximal stimulatory effect that

Secretion

PROLACTIN

MODULATES

PROGESTERONE

SECRETION

89
the effects combination secretion (Figs. 2C, effect (5to was 10-fold) of testosterone with testos-

was about three times the third day of culture.

greater

than

control

levels

by

In alone,

comparison prolactin

to in

DISCUSSION

terone increased the all days of culture that in pigs Lucky This of hormone et effect androgens culture ai., does androgen also but to FSH by et 1977; not to inand the from magnitude proestrous of rats the

of progesterone on 3C, 4C); however, greater than with with cells cells (2pro-

Previous enhance granuiosa al., Haney appear estrogen. creases secretion immature and Nimrod of tion cyclic of 1976; and to

research production cells Hillier from et

has of

demonstrated progesterone and 1977; 1978). rats

(Schomberg

al.,

from HPX to 5-fold)

rats cultured serum. Again, may from

with (2-fold) or without increased numbers of

Schomberg,

depend on aromatization Follicle-stimulating of interact progesterone,

lactin receptors response of cells three bination than The general (1982). hanced one the ours, In 4B), experimental of

be responsible for the greater proestrous rats. In each of the the effects of were hormone animals Wang that from was systems the by and the comgreater alone. are in Chan and testosterone

systems,

production

prolactin

testosterone

synergistically by granulosa with estrogen and that distal this to

increase

of progesterone HPX rats treated 1976; has FSH the that adenosine and Our two with increases AMP, (1977a,b) cyclic

cells from (Armstrong 1976). interaction the (AMP), cells reduction in 5 generathat to

the sum of the effects of each results for cells from proestrous agreement with those reported androstenedione cells increase doses of These authors the effects of secretion magnitude but all on they three by of used also the lower enhanced secretion

Dorrington, androgen

Nimrod reported occurs sensitivity it does not confirm and

Lindner,

prolactin enon progesterproestrous not (Figs. as great 2B, effect 3-fold. rats; as 3B, of This on all when without observed of cells the doses two of

granulosa

monophosphate affect

androgen

of granulosa

of prolactin. stimulatory 2- to

experimental

of progestins. effects of the stimulating or absence treated show that granulosa with

results hormones

these stimulatory their interaction cells in the from Our

prolactin

FSH

progesterone

progesterone production of serum by granulosa estrogen (Fig. 1A,C).

presence HPX rats also effects on (Fig. 1B).

enhancement of the days of culture and cells serum. lated until from hormones from HPX rats of and In contrast, secretion the second proestrous were

effects of FSH was evident was greater than additive were effects cultured of prolactin with on were not in cultures effects The of lowest or

results

androgen and FSH exert similar cells from proestrous follicles

FSH-stimu-

However, the thrust of these experiments was to examine the effect of prolactin, alone and in combination with testosterone and/or FSH, on the ability of granulosa results production, the cells to secrete progesterone in vitro. The show that prolactin affects progesterone but that the magnitude and, in one case, of its effects can be influenced by the hormones of the increased in the medium, granulosa cells, secretion the stage and the

progesterone third days and the additive.

rats, roughly

prolactin that FSH (0.02 or the one one or dose needed in control (2 pg/mi). As discussed in combination

significantly enhanced 0.2 pg prolactin/mi) to medium above, with all stimulate or in the the effects testosterone secretion

the effects of were lower than of progesterof testosteralone were Neither nor the influthese and

direction

presence

presence of other of differentiation culture conditions. In contrast to

of prolactin or FSH

of progesterone and proeswith proalone had

by cells from HPX (2- to 4-fold, Fig. 4A) trous rats (3- to 25-fold, Fig. 3A), cultured lactin in the presence of serum, prolactin little effect on one by granulosa fined from prolactin induces vivo and proestrous ably more medium HPX rats prolactin the negligible cells from production HPX rats

stimulatory under stage of differentiation presence enced factors or the did

conditions employed. of the granulosa cells in the medium effect, although of the response

of progestercultured in de-

absence of serum direction of the affect the magnitude

(Fig. may

2A). The lower response of cells be due to their limited number of (Wang receptors et on al., rat 1979). granulosa Since FSH cells in

the effective progesterone combination when prolactin HPX has

dose in some cases. Absolute accumulated with prolactin with FSH or testosterone cells been were cultured to with promote shown

amounts of alone or in were greater serum. Since cholesterol

receptors

in vitro (Wang rats would be prolactin.

et al., expected

1979), cells from to bind consider-

storage in rabbit ovarian tissue (Hilliard et al., 1968), it is possible that prolactin causes granulosa cells to increase their uptake of cholesterol or lipoproteins

90 present availability above, testosterone progesterone Prolactin testosterone proestrous in the serum of progestin to the in had in and thereby precursors. discussed with effects

FORTUNE

AND

VINCENT

increases in the both paragraph FSH

the

and et al., human

by

perifused

human

ovarian

fragments

(Demura

In contrast prolactin

results

1982). However, granulosa cells

in other studies with pig and both inhibitory and stimulatory alone
Ct

combination different

and of

different

on accumulation

effects of prolactin of serum (McNatty 1980). The effects clear. that the

were noted al., 1974;

in the Veidhuis

presence et al., exerts are reported estradiol its not

experimental

systems. of FSH + cells from (Figs. 3D, are in in the by greater FSH
+

enhanced the stimulatory effect on progesterone secretion by or HPX rats cultured with serum for cells stimulatory and (1982). of the

mechanism(s) on steroidogenesis Dorrington inhibitory and

by

which prolactin by granulosa of prolactin

cells on

Gore-Langton effects

(1982)

4D). The results accord with the presence Wang than and the of sum FSH Chan

from proestrous rats effect of prolactin reported was with This stimulation observed alone. seemed

androstenedione secretion

secretion seem to be exerted distal of cyclic AMP. However, it appears nism of its effects on estradiol may the mechanism progesterone; stimulated by which prolactin increases it modulates suppresses

to the formation that the mechabe different from production gonadotropinwhereas of

testosterone presence

alone and prolactin of serum, prolactin

Thus, in the to interact

in secretion

of estradiol

synergistically with the other two hormones to promote accumulation of progesterone. In contrast, prolactin inhibited secretion of progesterone in the presence of FSH + testosterone when cells from HPX rats were cultured in defined medium (Fig. 2D). Hence stage of alter the direction presence stimulatory obtained periments 1986). of serum), for granulosa cell of prolactins but absence differentiation did not effect (at least in the of serum reversed the differ from those in the same exFortune et inhibited occurs Therefore, under condiproduction it inhibited first in which of results in the al., the in

it may suppress or increase one. Possible mechanisms progesterone production cyclic olism lactin activity granulosa

production of prolactins are changes

of progesteractions on in levels of and in metabIn vivo, proincreases in in Nimrod, activity dispersed and

AMP, in precursor mobilization, of progesterone to other steroids. partially inhibits FSH-stimulated of cells 20a-hydroxysteroid of HPX rats (Eckstein

dehydrogenase

effect. These results estradiol secretion companion under all secretion FSH and paper; conditions,

1979); in vitro, prolactin of 20a-hydroxysteroid luteal cells from our experiments one may of (1982). In and
interact

appears to suppress dehydrogenase in rats (Wu concentration

(see Prolactin,

pregnant only one

et al., 1976). In of testosterthat Wang in prolactin and combination of and Chan

concentrations

increase in estradiol the presence of

that normally testosterone. secretion progesterone in which the

FSH

was used. differently as prolactin or

It is possible with different shown alone by or

prolactin inhibited estradiol tions in which it enhanced as well as under conditions progesterone This study the effects have that serum suggest production. is to our of prolactin been directly at least some may serum 10

these

hormones,

summary,

knowledge in the presence

with FSH progesterone differentiated 10% FSH terone; secretion which exerted

testosterone by relatively granulosa serum, to of cells.

increased secretion undifferentiated In medium containing

vs. absence

compared, and our of the contradictions be explained by in the culture

fetal bovine and testosterone in the of

prolactin interacted with increase secretion of progesserum, prolactin inhibited by are that The mechanism(s) and inhibitory effects determined. It is clear

previous literature or absence of medium enhanced secretion proestrous

the presence medium. In

absence

containing the stimulatory of progesterone rats (Wang and

or 15% serum, prolactin effects of gonadotropins on and by granulosa Chan, 1982; cells Grasso from and

progesterone. these stimulatory remains to be

Crisp, 1985), of progesterone 1980). In hCG-stimulated

prolactin alone by goat granulosa

increased secretion cells (Mohini et al.,

prolactin has dramatic effects on steroidogenesis by granulosa cells in vitro. At the same time, these results indicate that the interactions of prolactin with other hormones in vitro are complex, and extrapolations to situations in vivo must be made with caution.

serum-free secretion

medium, prolactin inhibited of progesterone by granulosa with hCG clomiphene (Soto et al., citrate 1985),

cells from women treated and hCG and cultured with

ACKNOWLEDGMENTS
We are grateful to the National Pituitary Agency, Baltimore, MD.

PROLACTIN for this supplying manuscript. ovine prolactin and FSH and to H.

MODULATES Dormady for typing

PROGESTERONE McNatty KP,

SECRETION Neal P, Baker TG, 1976. mouse AS, secretion on control Effect ovaries 1974. by fertility. A the of prolactin on

91
the

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