Archivesof
Oto-Rhino-Laryngology
9 Springer-Vcrlag 1978
Summary. Sodium, potassium, chloride, glucose and total protein were determined in samples of scala vestibuli perilymph, scala tympani perilymph, CSF,
cochlear endolymph and utricular endolymph from normal cats. Small but significant differences were evident in the concentrations of sodium and potassium
between the scala vestibuli and scala tympani perilymph. It was also apparent
that each compartment of endolymph has different values for sodium and potassium concentrations. Compared with the endolymph, the perilymph was found
to contain a higher concentration of glucose and total protein. These findings are
discussed from the view point of biological significance.
Key words: Inner ear fluid biochemistry - Micro-chemical analysis - Osmotic
pressure in inner ear fluids.
There is a paucity in knowledge concerning chemical composition in inner ear fluids.
In experiment series on inner ear fluid chemistry (Silverstein and Makimoto, 1973;
Makimoto and Silverstein, 1974), we analysed sodium, potassium, chloride, glucose
and total protein in samples of scala vestibuli perilymph, scala tympani perilymph,
cochlear endolymph, utricular endolymph and CSF. 125 ears in 95 cats served as
the materials. The present report is a summarization of the results heretofore obtained.
Sodium and Potassium
0302-9530/78/0220/0259/$ 01.20
260
K. Makimoto et al.
These results are shown in Table 1. As demonstrated here, small but significant
differences were evident between scala vestibuli and scala tympani perilymph both in
sodium and potassium concentrations. It was also apparent that each compartment
of endolymph has its own value for sodium and potassium concentrations.
In the perilymph of scala vestibuli, a higher concentration.of potassium was
noted. As to endolymph, a greater value was observed in the sodium concentration
of utricular endolymph. This m a y be regarded as one of the biological features for
utricular endolymph. In Figure 1, data on the concentrations of sodium and potassium in fluids o f extracellular type and serum are graphically illustrated. Regarding
sodium concentration, perilymph of the scala vestibuli was found to be closest to
serum. On the other hand, in potassium concentration, perilymph of the scala tympani showed the closest value to that of serum. When a comparison o f N a / K ratio
between scala vestibuli perilymph and serum was made, it was obvious that a greater
difference existed in this ratio. Accordingly, the perilymph of scala vestibuli m a y not
be a simple blood filtrate.
Chloride
Chloride was determined by the use of a Chloridemeter | With this instrument, the
mEq/1 of chloride in a sample can b e measured by passing a current through the
sample via two generating electrodes. The principle of this measurement is that silver
ions are released from the anode and combine with chloride ions in the sample. The
smallest amount required for this determination is 2.0 ~1.
Table 1. Sodium and potassium concentrations in inner ear fluids, CSF, and serum
Sodium
(mEq/1)
Potassium
Difference
(mEq/1)
Perilymph of
scala vestibuli (Pv)
Perilymph of
~
scala tympani (Pt)
Cochlear endolymph
(Ee)
Utricular endolymph
(Eu)
147.76 (+ 0 . 9 4 ) ~
(N = 54) ~ . > 9.35'
157.11'(+ 1.06) j ~ ( P t - - P v )
(N = 5 8 ) ~
6.07 (+ 0.40) ~
(N=41) " ~ 8 . 7 9 b
14.86 (+ 0.75) ~
(Eu-Ec)
(N = 44)
CSF
152.91 (+ 1.00)
(N = 32)
2.97 (+ 0.08)
(N = 32)
Serum
3.93 (+ 0.16)
(N = 27)
Difference
10.47 (+ 0.31) .
(N = 54) ~
6.68c
3.79 (+ 0.11) / / ~ (Pv--Pt)
(N = 58) :
171.13 (+ 1.64) .
(N = 41) ~ 1 5 . 7 0
a
155.43(+ 1.62) j
(Ec--Eu)
(N = 44) /
261
(rnEq/L)
160
150
140
130
I Sodium Concentration
~
Potassium Concentration
(N = 43) ~
130.00 (_+ 2.31)
(N = 46)
169.82 (+ 5.83)
(N = 28) ~
158.88 (+ 6.22)
( N = 24)
CSF
Serum
120.00 (+ 3.59)
(N = 30)
Difference
8.51 a (Pv--Pt)
10.94b (Ec--Eu)
262
K. Makimoto et al.
Glucose
Glucose determination was carried out by adapting the enzymatic test kit of Boehringer to a micro-chemical technique (Silverstein, 1973; Makimoto and Silverstein,
1974). The principle of this enzymatic method is as follows:
1. Hexokinase catalyses the phosphorylation of glucose by ATP.
Glucose + ATP -~ glucose-6-phosphate + ADP.
2. D-glucose-6-phosphate (G-6-P) is oxidized to 6-phosphogluconate (6-P-G) in
the presence of TPN (NADP) by the enzyme G-6-P dehydrogenase.
G-6-P + TPN -~ 6-PrG + T P N H + H +.
The amount of T P N H arising in Equation (2) is equivalent to the amount of G-6-P
or glucose, respectively.
The amount of the samples used in this measurement was 0.5 ~1 for endolymph
and 1.0 pxl for other fluids.
The results are presented in Table 3. The glucose concentration of cochlear
endolymph was appreciably low, being 11.2 mg/100 ml, whereas that of utricular
endolymph was higher, being 41.9 mg/100 ml. As previously mentioned, utricular
endolymph was found to contain a higher concentration of sodium as compared to
Table 3. Glucose concentrations in inner ear fluids, CSF, and Serum
Glucose
(rag/100 ml)
Cochlear endolymph (Ec)
Utricular endolymph (Eu)
Perilymph of scala Tympani (P0
Perilymph of scala vestibuli (Pv)
CSF
Serum
Difference
30.69a (Eu--Ec)
3.12b (Pt--Pv)
81.95 (+ 3.37)
(N = 49)
122.48 (+ 7.12)
(N = 45)
263
Total Protein
Total protein was determined by employing the Folin phenol reaction of Lowry et al.
(1951). As the first step of this method, the sample is treated with copper sulfate in
alkaline solution. Folin reagent is then added to the sample to develop the color
reaction.
According to the results shown in Table 4, the total protein contents of scala
vestibuli perilymph and scala tympani perilymph exceeded those of cochlear and
utricular endolymph. Also in CSF, the content of total protein was noted to be much
lower than those in scala vestibuli perilymph and scala tympani perilymph. This
pattern of concentration gradient in total protein did not coincide with that in glucose. The glucose study revealed that the glucose concentration of CSF was almost
the same level as those of scala vestibuli perilymph and scala tympani perilymph.
In Figure 2, the total amount of electrolytes (sodium plus potassium plus chloride) are illustrated on the left side of this figure. When this value is compared among
various compartments of inner ear fluid, the values for endolymph are greater than
those for perilymph.
233.87 (+ 1 5 . 7 0 ) ~
(N = 23) ~
212.52 (+ 1 0 . 6 6 ) ~
(N = 31)
61.50 (+ 7.49)
(N = 18)
64.33 (+ 7.70)
(N = 9)
37.89 (+ 6.24)
(N = 9)
Difference
a
21 35 (Pv--Pt)
"
2.83b (Eu-Ec)
264
K. Makimoto et al.
I0
[ p
I00
~.,e/ioo~
I mE;q/L
Pt
::::::::::::::::::Na
.........
( m t ::' q /-L ): : . :.:.:.:
.:. . . . .:.:.:-:
.]. . ..
C.S.F.
I:::::::::::::::: J:::;:::::::::::::::::::::::::::::::::?::::::::::]]
Eo
.-
iG'
<mEq,.
!ililll]01111NIllll01111111IIIllllllllllllll
III1
P v : Perilyrnph of Scala Vestibuli
Ec : Cochlear Endolyrnph
Eu : Utricular Endolyrnph
P r : Total protein (m E % )
GI : Glucose ( m g / I O O m l )
Fig. 2. Comparison of osmotic concentration among various compartments of inner ear fluid and
CSF
References
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin
phenol reagent. J. biol. Chem. 193, 265-275 (1951)
Makimoto, K., Silverstein, H.: Sodium and potassium concentrations in the endolymph and perilymph
of the cat. Ann. Otol. 83, 174--179 (1974)
Makimoto, K., Silverstein, H.: Effects of insulin on glucose concentrations in inner ear fluids and
cochlear microphonics. Laryngoscope 84, 722--737 (1974)
Silverstein, H.: Biochemical studies of the inner.ear fluids in the eat. Ann. Otol. 75, 48--63
(1966)
Silverstein, H., Schuknecht, H. F.: Biochemical studies of inner ear fluid in man. Arch. Otolaryng. 84,
395--402 (1966)
Silverstein, H.: Inner ear fluids in man. Laryngoscope 83, 79-107 (1973)
Silverstein, H., Makimoto, K.: Superior vestibular and "singular nerve" section. -- Animal and clinical
studies --. Laryngoscope 83, 1414--1432 (1973)
Received October 24, 1977