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PubMed2001

Effects of enzyme and anion transport inhibitors on in vitro incorporation of inorganic carbon and calcium into endolymph and otoliths in salmon Oncorhynchus masou.

The transepithelial transport of inorganic carbon to endolymph and its subsequent deposition on otoliths were pharmacologically examined by incubating the sacculus containing an otolith with NaH(14)CO(3). Calcium incorporation was also studied. Carbon incorporation into endolymph and otoliths was saturated with increased concentrations of bicarbonate ions in the incubation medium and was followed by the Michaelis-Menten equation with a K(m) of 26.3 mM and 0.4 mM, respectively. Carbon incorporation decreased with an increase in chloride concentrations in the medium. Calcium incorporation was not affected by chloride and bicarbonate ions up to 10 mM. Higher concentrations of bicarbonate ions reduced calcium incorporation into both fractions. Carbon incorporation into endolymph and otoliths was inhibited by acetazolamide, disulfonate stilbenes (DIDS and SITS), thiocyanate, and ouabain. Calcium incorporation was not affected by these inhibitors. Amiloride inhibited carbon incorporation into otoliths alone. These results suggest that HCO(3)(-)-ATPase and Cl(-)/HCO(3)(-)-exchangers are involved in the transepithelial transport of bicarbonate ions to the endolymph. Carbonic anhydrase was also suggested to play a role in carbonate production for otolith calcification.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1989

[Flow kinetics of endolymph in rotational stimulation].

In this paper, experimental and mathematical results are presented, indicating rotatory nystagmus to be caused by endolymph flow in the ampulla of the horizontal semicircular canal. During positive or negative acceleration circular flows may develop within the endolymph system, especially in the ampulla of the horizontal semicircular canal. While being independent of the position of the axis of the horizontally rotating system, the velocity of the flow depends on the radius of the establishing "whirl" in the ampulla. It is demonstrated that, close to the corresponding angular acceleration threshold, the velocity of the tangential flow along the cupula is in the same range as that calculated for a caloric stimulus. Like in our model of calorisation also in rotation there is strong evidence that an endolymph flow directed from crista to cupula in the lateral part of the ampulla causes a nystagmus towards the corresponding side, whereas a flow from cupula to crista causes a nystagmus in the opposite direction. Our results suggest that endolymph flows, evoked either by calorisation or rotation, represent adequate stimuli for vestibular excitability.

Acceleration↗
PubMed1989

Lack of effect of carbonic anhydrase inhibition on direct measurements of endolymph bicarbonate.

In the past, sodium, potassium, and chloride have been measured in endolymph directly, but bicarbonate has been measured only indirectly. We sampled endolymph directly while monitoring endocochlear potentials in normal and methazolamide-treated guinea pigs. Bicarbonate was determined in samples by use of a method that depends on reduction of NADH to NAD linked to malate formation from oxaloacetate. In 11 normal animals, the bicarbonate in endolymph was 20.2 mM +/- 4,4 mM (mean +/- standard deviation); in six of these, plasma bicarbonate was 23.1 mM +/- 3.5 mM. Nine animals treated with methazolamide (carbonic anhydrase inhibitor) had an endolymph bicarbonate of 19.5 mM +/- 3.9 mM; plasma bicarbonate in five of these was 25 mM +/- 3.2 mM. Carbonic anhydrase inhibition did not significantly affect endolymphatic bicarbonate levels.

Animals↗
PubMed1981

Steep gradients of amino acids between cochlear endolymph and perilymph.

Levels of 19 free amino acids in cochlear endolymph and perilymph of scala vestibuli of the guinea pig were determined using reverse phase high performance liquid chromatography (HPLC) of the o-phthaldialdehyde-ethanethiol derivatives with fluorescence detection. Aspartate and glutamate were found to be significantly higher in endolymph than in perilymph, confirming results from another study using a different analytical method. The remaining 17 amino acids were significantly lower in the endolymph, in many cases by an order of magnitude. The data are compared with results on utricular endolymph and vestibular perilymph obtained by HPLC in another study from our laboratory. Possible implications and interpretations of the results are discussed.

Amino Acids↗
PubMedDate not supplied

Pressure relationship between perilymph and endolymph in guinea pigs.

The pressure difference between the perilymph and the endolymph was studied in guinea pigs, using two sets of a servo-nulling system. Asphyxia caused characteristic pressure changes both in the perilymph and in the endolymph, but no pressure difference was seen between them. When the perilymphatic space was opened to the atmosphere, asphyxia did not cause such dynamic pressure changes either on the perilymph or the endolymph. An increase in the perilymphatic pressure induced by the infusion of artificial cerebrospinal fluid into the subarachnoid space resulted in an increase in the endolymphatic pressure, but no pressure difference between the endolymph and the perilymph was evident, regardless of the patency of the vestibular aqueduct. The authors conclude that the endolymphatic pressure is dependent on the perilymphatic pressure and there is no pressure difference between them.

Animals↗
PubMed1993

[Contribution of cell culture to the study of cochlear endolymph].

Investigation of the mechanisms involved in the secretion of cochlear endolymph is hindered by the heterogeneity, the fragility, and the difficulty of access to the secretory structures of the inner ear, i.e. stria vascularis and Reissner's membrane. Cell culture may overcome these limitations and allow the study of the epithelial transport processes. Cultured cells derived from gerbil's explants of stria vascularis and Reissner's membrane presented morphological and immunohistochemical properties similar to those of the native cells. Investigation by the patch clamp technique of the apical membranes of marginal-like cells showed the presence of non specific cationic channels that could allow passive K+ flow towards endolymph and, thus, play a major role in the production of endolymph. Biochemical studies demonstrated that a cAMP production by these marginal-like cells. Primary culture of stria vascularis and Reissner's membrane is a suitable model to further investigate the physiology of endolymph production.

Animals↗
PubMed2002

Optimization of the mechanical performance of a two-duct semicircular duct system--part 2: excitation of endolymph movements.

The endolymph flow inside the semicircular ducts is analytically investigated by considering a system of two hydrodynamically interconnected ducts. Rotation of this system adds an amount of motion (momentum) to parts of it. This results in an endolymph flow in generally all vestibular parts. The "external impulses" are the impulses which emerge by rotation of exclusively a particular vestibular part. The real impulses can be calculated from a set of equations which contain the external impulses. Analytical expressions are derived for the initial velocities in the ducts and for the maximum endolymph displacements. These formulae contain the external impulses and the ratios of: (1) the radii of crus commune and ducts (gamma), (2) the lengths of crus commune and ducts (lambda). It was proven that an interconnected system composed of two ducts, and also a system composed of two such semicircular duct systems, behaves as a pure rotation transducer (like a single duct does), also when it is rotated excentrically. Duct systems with polygonal and circular geometries were used to evaluate whether an optimal value of lambda would exist (gamma was already considered elsewhere). Optimum values of lambda in a range of about 0.10-0.52 were found. This rather wide range of values agrees with values from measurements. Optimization of an interconnected duct system appeared to be equal to optimization of a system composed of separate ducts.

Animals↗
PubMed1988

Magnesium ion activity in the mammalian endolymph measured with ion-selective microelectrodes.

The free Mg++ concentration in endolymph was measured with Mg++-selective microelectrodes based on the neutral ligand ETH 1117. The property of Mg++ microelectrodes was obtained from calibration solutions, containing various Mg++ concentrations with the background electrolytes resembling endolymph. The range between 10 and 0.1 mM Mg++ concentrations changed the potentials of Mg++ microelectrodes by 14.4 +/- 3.0 mV. The endocochlear potential and the Mg++ concentration in the endolymph were 82.0 +/- 5.0 mV and 0.77 +/- 0.29 mM in the guinea pig, and 84.4 +/- 4.9 mV and 1.12 +/- 0.24 mM in the chinchilla, respectively. These results are discussed in the light of the dependence of Na+, K+-ATPase and its interaction with Ca++.

Animals↗
PubMed2003

Composition of biomineral organic matrices with special emphasis on turbot (Psetta maxima) otolith and endolymph.

The soluble organic matrix (OM) of various biominerals (red coral skeleton, oyster shell, sea urchin test, turbot otolith, chicken eggshell) was extracted after demineralization with acetic acid. The protein content of the OM varies strongly from 0.02 to 1.6 microg/mg biomineral whereas proteoglycans present less variations (from 0.7 to 1.4 microg/mg biomineral). Electrophoresis of biominerals OM shows differences in their protein pattern although several bands are present in all matrices. OM of all biominerals shows carbonic anhydrase activity but no activity was detectable in the endolymph. OM of all biominerals also displays an anticalcifying activity. After separation of the OM extracts by chloroform-methanol, 80% of the anticalcifying activity was found in the methanol phase except in the urchin test. After OM precipitation with trichloracetic acid, 70% of the activities was found in the supernatants. Partial biochemical characterization suggests that the anticalcifying factor is a polyanionic and water-soluble molecule, which could be proteoglycans. The endolymph surrounding the otolith also displays an anticalcifying activity although its inhibitous activity was 50 times lower than that of the otolith OM. However, the anticalcifying activity of the endolymph is assumed by a proteic structure (80% activity precipitated with TCA treatment). Our results suggest that both carbonic anhydrase and anticalcifying activities are widespread and play a significant role in the regulation of biomineral formation. Results are discussed in relation to the calcification process that takes place at the fluid-mineral interface.

Animals↗
PubMed1978

A correlation of the effects of normoxia, hyperoxia and anoxia on PO2 of endolymph and cochlear potentials.

Change in PO2 in endolymph, endocochlear potentials and cochlear microphonics have been tested in normoxia, hyperoxia and anoxia on 24 guinea pigs. The polarographic method and construction of oxygen-sensitive microelectrodes is described in detail. The normal level of PO2 in endolymph vaires between 20 and 30 mm Hg. One-minute anoxia induced by breathing 100% N2 caused a decline in PO2, EP and CM, but during recovery only PO2 returned with over-correction to the preexposure level. Hyperoxia evoked by breathing 100% oxygen failed to increase the cochlear potentials and the PO2 in the endolymph. This suggests that vasoconstriction most likely occurs proximal to the capillaries bed of the stria vascularis.

Animals↗
PubMed1983

Ethacrynic acid facilitates gentamicin entry into endolymph of the rat.

Influence of ethacrynic acid (EA) upon gentamicin kinetics in perilymph and endolymph was studied in rats that were given a constant-infusion of gentamicin (150 micrograms/min) and EA (140 micrograms/min). Inner ear fluids and plasma were sampled up to 5 h. The purity of the endolymph was ensured by measurement of sodium and potassium concentrations. Gentamicin assay was done with a modified radioimmunoassay. Results show that EA facilitates the entry of gentamicin into endolymph, while it does not affect the kinetics of the drug in perilymph. Although the mechanism of this facilitation remains unclear, this result may account for the ototoxic potentiation reported between EA and aminoglycoside antibiotics.

Animals↗
PubMed1984

Water permeability of the endolymph-perilymph barrier in the guinea pig cochlea.

The diffusional water permeability of the endolymph-perilymph barrier was determined in guinea pigs by perfusing the perilymphatic space with tritiated water and measuring the uptake of tritiated water into the endolymph. Assuming that the volume flow across the endolymph-perilymph barrier is negligible for short periods of perfusion, the water permeability of the barrier is approximately 130 times greater than its permeability to K+ in normal guinea pigs.

Animals↗
PubMed1986

Changes in endolymph chloride concentration following furosemide injection.

Endocochlear potential (EP) and chloride concentration in endolymph were monitored with microelectrodes in the basal turn of the cochlea of the chinchilla. After intravenous injection of furosemide (25-100 mg/kg), the EP dropped precipitously and rapidly reached its minimum value, however, the chloride activity in endolymph decreased more gradually. Possible mechanisms for this phenomenon include a reduced electrostatic attraction of chloride ions to the scala media due to a decreased EP and a reduction of passive influx of chloride into endolymph, resulting from a reduction of active inward potassium transport by furosemide.

Animals↗
PubMedDate not supplied

Ionic changes in cochlear endolymph of the guinea pig induced by acoustic injury.

The effects of acoustic overstimulation on the endocochlear potential (EP) and on concentrations of ions (K+, Na+, Cl-, H+, HCO3-, and Ca2+) in endolymph were investigated using ion-selective microelectrodes. A slight but significant elevation of the EP and alkalinization of the endolymph were induced by acoustic overstimulation, whereas there was little change in the K+, Na+, Cl-, and HCO3- concentrations. The changes in H+ and HCO3- concentrations implied a depression of PCO2, suggesting an increase in blood flow to the cochlea. On the other hand, the Ca2+ concentration increased abruptly to 48 times the pre-exposure value. In contrast, no significant change in the Ca2+ concentration was observed in cochleae with damaged hair cells. We discuss the mechanism of the tone-induced Ca2+ elevation in endolymph and its effect on hearing acuity.

Animals↗
PubMed1992

Protein profiles of perilymph and endolymph of the guinea pig.

Results of protein separation of guinea pig plasma, perilymph, and endolymph by means of high-resolution two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis are presented. Several proteins are present in perilymph at levels in basic accord with the total protein gradient with respect to plasma; however, others are present in perilymph at levels comparable to plasma levels, and one protein low molecular weight protein, PLS:33, is eight times higher. In addition, a high molecular weight protein is shown to be present at similar levels in the two compartments. These findings indicate that ultrafiltration cannot be the sole mechanism of perilymph production. Endolymph proteins are uniformly five to eight times lower than perilymph levels, essentially following the total protein concentration gradient between the two compartments. This supports the view that endolymph is derived from perilymph rather than directly from blood.

Animals↗
PubMed2006

EphB2 and ephrin-B2 regulate the ionic homeostasis of vestibular endolymph.

The ability to transport cations and anions across epithelia is critical for the regulation of pH, ionic homeostasis, and volume of extracellular fluids. Although the transporters and channels that facilitate ion and water movement across cell membranes are well known, the molecular mechanisms and signal transduction events that regulate these activities remain poorly understood. The Eph family of receptor tyrosine kinases and their membrane-anchored ephrin ligands are well known to transduce bidirectional signals that control axon guidance and other cell migration/adhesion events during development. However, these molecules are also expressed in non-motile epithelial cells, including EphB2 in K(+)-secreting vestibular dark cells and ephrin-B2 in the adjacent transitional cells of the inner ear. Consistent with these expression patterns, mice with cytoplasmic domain mutations that interfere with EphB2 forward signaling or ephrin-B2 reverse signaling exhibit a hyperactive circling (waltzing) locomotion associated with a decreased amount of endolymph fluid that normally fills the vestibular labyrinth. Endolymph is unusual as an extracellular fluid in that it is normally high in K(+) and low in Na(+). Direct measurement of this fluid in live animals revealed significant decreases in K(+) concentration and endolymphatic potential in both EphB2 and ephrin-B2 mutant mice. Our findings provide evidence that bidirectional signaling mediated by B-subclass Ephs and ephrins controls the production and ionic homeostasis of endolymph fluid and thereby provide the first evidence that these molecules can control the activities of mature epithelial cells.

Animals↗
PubMed1999

Sodium, potassium, chloride and calcium concentrations measured in pigeon perilymph and endolymph.

According to Davis' (1965) model of the inner ear, a potential difference between the endocochlear potential and the hair cell resting potential drives the transduction current across the apical hair cell membrane. It is assumed that the endocochlear potential (EP) consists of two components. The first is a diffusion potential, which depends on the ionic composition of endolymph and perilymph and on the permeability of the perilymph-endolymph barrier. The second is an electrogenic component which is determined by active ion transport across the perilymph-endolymph barrier. In birds, the EP is between +8 and +20 mV. Little is known about the underlying mechanisms responsible for the measured EP in birds. The present paper studies whether ionic compositions of endo- and perilymph might explain the EP in birds. Concentrations of Na+, K+, Ca2+ and Cl- in pigeon scala vestibuli, scala tympani and scala media were determined with ion-selective microelectrodes. Na+, K+, Ca2+ and Cl- were 150.0, 4.2, 1.4 and 117.0 mM in perilymph (scala tympani and scala vestibuli). In scala media, the concentrations of K+, Ca2+ and Cl- were 140.6, 0.23 and 142.1 mM.

Animals↗
PubMed2001

Vesicular storage of adenosine triphosphate in the guinea-pig cochlear lateral wall and concentrations of ATP in the endolymph during sound exposure and hypoxia.

Previous studies have revealed putative vesicular stores of adenosine triphosphate (ATP) in the marginal cells of the cochlear stria vascularis which may serve as a source of ATP for purinergic signalling. This study aimed to provide further evidence of ATP storage in the cochlea and to see whether ATP levels in the endolymph are affected by noise and hypoxia. Tissues from the lateral wall and organ of Corti of the guinea-pig cochlea were fractionated to obtain vesicular (VF) and mitochondrial (MF) fractions. Free and total ATP were then measured by the luciferase-luciferin reaction from which membrane-bound vesicular ATP was calculated. In the lateral wall, the VF contained 2.02+/-0.04 nmol ATP/mg protein (n = 5), significantly greater (p < 0.001; paired Student's t-test) than the concentration of ATP in the MF (0.36+/-0.05). In the organ of Corti, the VF contained 0.69+/-0.08 nmol ATP/mg protein (n = 4), significantly smaller than the amount in the VF of the lateral wall tissues (p < 0.001; non-paired Student's t-test). Small amounts of fumarase. an enzyme of the mitochondrial matrix, in the VF, excluded the possibility of mitochondrial ATP contamination. To investigate the effect of hypoxia and noise on the ATP concentrations in the endolymph, fluid samples were collected from the first (basal) cochlear turn of anaesthetized guinea-pigs. As a result of hypoxia (15 min, 13% F1O2), ATP concentrations (nM, mean +/- SEM) increased from 6.2+/-2.3 to 9.3+/-4.5 (n = 4), but the difference was not statistically significant. As a result of noise (15 min, 10 kHz, 110 dB SPL. broad band), the ATP levels increased significantly from 7.4+/-1.2 to 16.0+/-1.8 (p = 0.01; Student's t-test: n = 4). This study has demonstrated the presence of a vesicular store of ATP in the stria vascularis of the cochlea and described an increase in the ATP levels in the endolymph during noise exposure. The findings suggest that ATP is actively secreted from the vesicular store under conditions of metabolic stress. The presence of ATP under basal conditions supports a role for ATP in the sound transduction process during normal function.

Adenosine Triphosphate↗