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At least 109 records · Page 6Linked to original sources

Alterations in oxygenation of cochlear endolymph during loud sound exposure.

The oxygen tension (pO2) of endolymph of the guinea pig cochlea was measured during exposure to loud sound (12 kHz or high-pass noise; 110 dB SPL up to 1 h duration). A small, but significant, steady decline in mean pO2 was observed after both pure tone and high-pass noise exposure. The extent of the change in pO2 varied from 0-50% in individual animals, compared with unexposed control animals. All exposed animals had an extensive loss of compound action potential (CAP) thresholds at frequencies of 8-30 kHz. However, there was no relationship between the extent of the change in pO2 of endolymph and CAP threshold loss.

Animals↗

Fluid kinetics of endolymph during rotation.

Rotatory vestibular excitability is discussed on the basis of a fluid-mechanical theory. The principle of this theory, already described for caloric excitation, is that appropriate stimulus causes flows within the endolymph. Experimental and mathematical approaches indicate these flows to be essentially the same in caloric as in rotatory stimulation. The direction of a horizontal nystagmus is apparently determined by a circuit of endolymph in the lateral part of the ampulla.

Endolymph↗

Effect of glycerol on pressure difference between perilymph and endolymph.

The pressure difference between the perilymph and the endolymph following administration of 50% glycerol (12 ml/kg) was studied in guinea pigs. The perilymphatic and endolymphatic pressures were measured simultaneously with two sets of a servo-nulling system. Glycerol was administered for about one minute via a gastrocatheter. Both the perilymphatic and endolymphatic pressures began to decrease about 5 min after the administration of glycerol, and thereafter the decrease continued for about one hour with no significant difference between the two pressures. We concluded from the results that the glycerol-induced pressure difference between the perilymph and the endolymph, if present, is only a very small one, although a collapse of the scala media after glycerol intake was reported in guinea pigs and chinchillas.

Animals↗

Convection, buoyancy or endolymph expansion: what is the actual mechanism responsible for the caloric response of semicircular canals?

The mechanisms underlying caloric nystagmus are still matter of debate. The original theory proposed by Barany and more recently by Pau and Limberg suggested that convective endolymphatic currents were involved. In contrast Gentine et al. suggested that the main mechanism responsible for caloric nystagmus is buoyancy due to calorization of the endolymph, without the need of continuous convective currents. Finally, other authors (Scherer and Clarke, Arai et al.) proposed that thermal expansion or contraction of the endolymph were involved. In the present study experimental conditions have been considered able to discriminate between these different models. The experiments, were carried out on isolated labyrinth preparations of the frog. Only the predictions of the model based on buoyancy were fully consistent with the experimental results whereas those provided by the other models were not.

Animals↗

[Ion-selective microelectrodes: principle and application of in vivo measurements of ionic concentrations in cochlear endolymph].

OBJECTIVE: To introduce the method of making neutral carrier ion-selective microelectrode and use it for in vivo measurement of ionic concentrations in cochloear endolymph. METHODS: GG-17 glass capillaries with 1.85 mm O.D. were rinsed extensively and dried in an oven. Two capillaries were parallelly connected and pulled with microelectrode puller to make a double-barreled micropipette. One barrel was back-filled with ion exchanger and internal reference solution, which served as ionic potential electrode, while the other barrel filled with 150 mmol/L KCL as reference electrode. Each barrel of microelectrode was connected to differential electrometer via Ag-AgCl wire and output was recorded on a three-channel recorder. Every ion-selective microelectrode was calibrated in a series of standard solutions to determine the required characteristics. Twenty healthy guinea pigs with normal hearing were anesthetized and were artificially respired through the tracheal canal after the intramuscular injection of suxamethonium chloride. The tympanic bulla was exposed and a double-barreled ion-selective electrode was inserted into the scala media through the round window and basilar membrane. The ionic potentials and endocochlear potentials (EP) were simultaneously recorded from the basal turn of the cochlea. Ionic concentrations were then calculated by Nicolsky-Eisenman equation. RESULTS: The concentrations of potassium, sodium and calcium ions in the cochlear endolymph were 146.3 +/- 11.8 mmol/L, 0.36 +/- 0.22 mmol/L and 16.2 +/- 5.7 micromol/L, respectively. CONCLUSIONS: The neutral carrier ion-selective microelectrode made with the above method is reliable and therefore can be used for accurately measuring the ionic concentrations of microenvironment in vivo continuously and transiently.

Animals↗

[Numerical modeling of convection diffusion of endolymph in porous structures of the vestibular apparatus].

The mathematical algorithm of the diffusive model of vestibular reception is discussed. The conditions to reduce arising 3-dim boundary value problem to 1-dim one are found. And it is shown, that separate diffusive canaliculi as a mechanical transducers demonstrate the apparently clear properties of the frequency selectivity relating to their transduction of the endolymph space pressure into the endolymph concentration changes in their outputs. Some numerical experimental results are presented and discussed.

Diffusion↗

Mechanisms of endolymph secretion.

In the inner ear, the membranous labyrinth, a tight heterogeneous sensory epithelium, separates two compartments that are filled with fluids of completely different composition. The lumen of the membranous labyrinth is filled with endolymph, a K-rich, positively polarized fluid, whereas the surrounding spaces are filled with perilymph, with a composition similar to an usual extracellular fluid. The inner ear fluids play a major role in the cochlear and vestibular physiology by the transmission of the mechanical stimulus to the hair cells, on the one hand, and by the transduction of this signal to a nerve potential, on the other hand. Numerous studies have been performed in order to know the chemical and physical characteristics of the inner ear fluids. A high, positive transepithelial potential has been evidenced in the cochlea together with a high K concentration and a low Na concentration. During the last years, the composition of the inner ear fluids, the origins of endolymph and perilymph, and the cellular mechanisms involved in the secretion of these fluids have been a great part elucidated. The present paper is a review of the contribution of the lab to the understanding of the physiology of the inner ear.

Animals↗

Physiology and chemistry of cerebrospinal fluid, aqueous humor and endolymph in Squalus acanthias.

By means of the appropriate isotopes injected into the spiny dogfish, Squalus acanthias, the transfer of all major ions into cerebrospinal fluid (CSF), aqueous humor (A) and endolymph (E) was studied. In addition, the effect of raising pCO2 in sea-water upon HCO3- concentration of these fluids was measured. In the several types of experiments, acetazolamide or methazolamide was used to inhibit completely carbonic anhydrase. The rates of fluid formation and ion transfer in CSF and A were fairly close, but those for E were far slower. The general pattern of ion transport in the three fluids were the same, Na+ (or Na+ + K+ in E) entry greater than Cl - entry, and the difference was HCO3-. The greater rate constants for HCO3-, increase in its entry rate by elevation of pCO2, and inhibition of its appearance by the sulfonamides, show that this is a special case of transport; the ion is formed in secretory cells from gaseous CO2 + OH-. Secretory cells at sites of formation of all the fluids contain both carbonic anhydrase and Na+-K+-ATP-ase, which subserve HCO3- formation and Na+ (or K+) transport. Comparison of these results with studies in mammals show that the vertebrate pattern for secretion of these three fluids is well established in the elasmobranch.

Adenosine Triphosphatases↗

Endocochlear potential and potassium concentration in endolymph and perilymph of the chinchilla.

Guinea pigs and chinchillas were studied for EP and potassium concentrations in scala media and scala tympani using potassium-sensitive microelectrodes. Response of EP to 3 min anoxia was strikingly different in these two species. On the other hand, the resting values for EP and potassium concentrations in endolymph and perilymph were not significantly different. These findings suggest that the different response to anoxia in these two species is due to differences in permeability of the cochlear partitions to the ions.

Animals↗

Effects of noise on cochlear potentials and endolymph potassium concentration recorded with potassium-selective electrodes.

Guinea pig cochleas were exposed to either broad-band noise at intensities between 95 and 115 dBA or octave-band noise centered at 380 Hz or 4.2 kHz at intensities between 115 and 125 dB SPL. Cochlear microphonics (CM), summating potentials (SP) and action potentials (AP) were recorded from differential electrodes in the perilymphatic scalae between successive 20-min periods of noise exposure. The endocochlear potential (EP) and endolymph potassium concentration [Kendo+] were recorded continuously from scala media using double-barreled potassium-sensitive electrodes. It was found that the initial exposure to noise at 115 dBA produced considerable suppression of the CM and AP, while the EP and [Kendo+] were elevated above their normal values. When animals previously treated with kanamycin were subjected to the same level of noise exposure no systematic increase in either EP ro [Kendo+] was observed. After prolonged exposure to 380 Hz octave-band noise at 125 dB SPL, a slow decline of EP and [Kendo+] was observed. The relationships between the changes in EP, [Kendo+] and CM are discussed.

Animals↗

Longitudinal distribution of cochlear potentials and the K+ concentration in the endolymph after acoustic trauma.

Guinea pigs were exposed to 142 dB third-octave band of noise control at 1 kHz for 1 h. At different times after exposure the endocochlear potential (EP), the anoxic negative endocochlear potential (-EP), the concentration of K+ (K+e) and microphonic potentials were recorded in scala media in four cochlear turns. The remaining hair cells were counted in each animal. Immediately after the exposure, the EP and K+e decreased evenly in all four cochlear turns and gradually returned to normal physiological values in 5-20 days. When measured 20 days after the exposure, essentially normal EP and K+e values were observed, with an apicalwards decline, which was similar to that found along the cochlea in nonexposed animals. Abnormal increased EP was observed in some animals 20 days after the exposure in the first and second turns. In contrast to positive EP and K+e values, the anoxic negative EP attained less negative values in the second turn of exposed animals, i.e., in the turn where the narrow band noise exerted the major destructive effect. An almost normal distribution of hair cells and most negative EP values were found in the fourth turn. The distribution of persistent hair cells correlated positively with the values of the anoxic negative EP and amplitudes of the microphonic potentials. It is assumed that, in addition to the difference in K+ concentration between endolymph and perilymph, the anoxic negative EP is dependent upon the functional state of the organ of Corti.

Animals↗

The junctions of the spindle-shaped cells of the stria vascularis: a link that completes the barrier between perilymph and endolymph.

It is current opinion that the intercellular spaces of the stria vascularis represent a closed compartment isolated from the endolymph by the tight junctions of the marginal cells and from the perilymph by the junctional complexes of the basel cells. However, it has not yet been investigated whether these two barriers meet at the stria margins toward Reissner's membrane and the spiral prominence. Possible candidates for this sealing could be junctions between the spindle-shaped cells. In the present study freeze-fracture replicas of guinea pig specimens fixed in the presence of filipin were used in order to investigate the junctions of the spindle-shaped cells and to localize the cholesterol in their plasma membrane. Replicas reveal that, below the belt-like apical zonula occludens, the basolateral plasma membranes of the spindle-shaped cells adjacent to each other and to the basal cells are joined over their entire extension by a large number of junctional strands intermingled with numerous filipin-cholesterol-complexes. Gap junctions are present in the meshes formed by these junctional strands. Thus, the plasma membrane of the spindle-shaped cells shows morphological and cytochemical characteristics which indicate that they are the anatomical components completing the barrier isolating the intrastrial compartment from the surrounding fluids.

Animals↗

Endocytosis and transepithelial transport of endolymph in the endolymphatic sac.

The fate of cationized ferritin (CF) injected into the endolymphatic space of the endolymphatic sac was observed by transmission electron microscopy. At 10 min after the injection, CF particles bound to the apical plasma membrane of epithelial cells of the sac and were then endocytosed with coated pits. However, they never passed through the junctional complexes between the epithelial cells. At 30 min after the injection, the CF particles were transferred to endosomes and lysosomes by small vesicles of 100-150 nm in diameter. CF particles were also found in small vesicles close to Golgi cisternae and in multivesicular bodies. Acid phosphatase positive lysosomes were found close to endosomes containing CF particles. In addition, a small fraction of the small vesicles containing CF particles became inserted into the basolateral plasma membrane. At 60 min after the injection, many CF particles were found in acid phosphatase positive secondary lysosomes. These observations suggest that endocytosis of endolymph is actively performed by the epithelial cells of the sac, and transepithelial vesicular transport across the epithelial cells occurs.

Acid Phosphatase↗

Adenosine 5'-triphosphate (ATP) concentrations in the endolymph and perilymph of the guinea-pig cochlea.

The concentration of adenosine 5'-triphosphate (ATP) in endolymph (EL), perilymph (PL) and cerebrospinal fluid (CSF), collected from anesthetized guinea pigs was determined using the luciferase-luciferin reaction. The cochlea was exposed by a ventrolateral approach and the bone overlying scala media of the third turn (EL) or scala vestibuli of the first turn (PL) was shaved to a thin layer and a small fenestrum made. For EL sampling, a double-barrelled pipette was inserted through the spiral ligament-stria vascularis complex. One barrel was filled with 150 mM KCl to record the endocochlear potential (EP) and upon the appearance of the positive EP, 0.12-1.22 microliter of fluid was aspirated into the other barrel by gentle negative pressure. For PL sampling, a single-barrelled pipette was advanced into scala vestibuli and 0.3-1.6 microliter of fluid was collected by capillarity. CSF (0.36-1.75 microliter) was obtained from the cisterna magna. The cochleae were removed and processed for light microscopy to determine the extent of tissue damage from the sampling procedure. ATP concentrations (mean +/- SEM, nM) for EL, PL and CSF were 12.95 +/- 2.4 (n = 10), 10.5 +/- 3.9 (n = 11) and 16.1 +/- 5.4 (n = 11) respectively. Differences in ATP concentrations among fluids were not statistically significant. To test the effect of hypoxia on ATP levels, a group of guinea pigs was subjected to a 90 s period of respiratory anoxia prior to sampling of EL, PL or CSF. ATP concentrations were 14.4 +/- 3.5 (n = 11), 20.7 +/- 4.1 (n = 10) and 13.5 +/- 4.6 (n = 4) for EL, PL and CSF, respectively; only PL ATP concentrations were statistically different (P = 0.018, Wilcoxon rank sum test) to basal conditions. This is the first study which demonstrates the presence of free ATP in cochlear fluids. The results indicate that ATP is present in cochlear fluids at concentrations close to those known to cause hair cell depolarization in vitro.

Adenosine Triphosphate↗

Does the endolymph pass through the base of the cupula?

Whether the endolymph of the semicircular canal passes the cupular partition or not was examined using the lateral semicircular canal system of adult pigeons (Columba livia). By applying various pressures by means of injection of a dye solution through the membranous canal, it was found that the dye solution, was seen to pass the cupula even under very low pressures when the pressure was increased gradually. When pulled by a magnet, the ultrafine particles of the dextran magnetite contained in the injected fluid were found to pass through the subcupular space without evident increase of the ampullary pressure.

Animals↗

Hydrostatic pressure measurement of endolymph and perilymph in the guinea pig cochlea.

Hydrostatic pressures of endolymph and perilymph were measured with a servo-micropipet system. The validity of the experiment was assessed by observation of endolymphatic pressure changes during and after 3 minutes of anoxia. Simultaneous recording of endocochlear DC potential confirmed the location of the electrode tip. Following verification of the efficacy of the system, measurements of endolymphatic or perilymphatic pressures were made in ten guinea pigs in which the middle ears had been filled with a saturated sodium chloride solution. Both endolymphatic and perilymphatic measurements showed gradual declines in pressure, with minimum pressure at 15 to 20 minutes and slow recovery during the next 20 minutes.

Animals↗

Fixation-induced shrinkage of Reissner's membrane and its potential influence on the assessment of endolymph volume.

The quantification of endolymph volume by histological techniques or by magnetic resonance (MR) microscopy requires the inner ear to be first treated with chemical fixatives. If the fixative induces soft-tissue shrinkage, it would tend to return a distended Reissner's membrane towards a straight position, since this membrane is anchored to bone at its medial and lateral edges. The goal of this study was to determine the degree of Reissner's membrane shrinkage induced by different fixation protocols to establish methods which minimize tissue shrinkage. Fragments of fresh Reissner's membrane were dissected from isolated cochleae in an artificial perilymph. Specimens were viewed with an inverted microscope during infusion of fixatives, and changes recorded on video tape. Size changes of the specimen were quantified, usually over a 20 min period. Heidenhain-Susa, a fixative which is widely used in histological studies of hydropic cochleae, caused substantial shrinkage of Reissner's membrane, decreasing the length of specimens by an average of 15.1%. Other fixation procedures induced far less shrinkage. The use of 3.1% glutaraldehyde in Hanks' balanced salt solution produced a mean length decrease of only 0.3%. The inclusion in the fixation medium of 4.5% mercuric chloride, corresponding to the concentration which is present in Heidenhain-Susa and which acts to increase the contrast of Reissner's membrane in MR microscopy, contributes significantly to specimen shrinkage. We can conclude that the degree of endolymphatic hydrops may be underestimated in specimens fixed with media containing high levels of mercuric chloride.

Animals↗

P2X receptor-mediated changes in cochlear potentials arising from exogenous adenosine 5'-triphosphate in endolymph.

Our previous studies have determined the presence of adenosine 5'-triphosphate (ATP) in the cochlear fluids and shown that extracellular ATP introduced into the endolymphatic compartment of the guinea pig cochlea has a significant dose-dependent suppressive effect on both endocochlear potential (EP) and cochlear microphonic (CM), which is mediated via P2 receptors. In the present study, the influence of P2 receptor agonists and antagonists on this suppressive effect was investigated to characterise the subtypes of P2 receptor mediating the ATP-induced effect on cochlear function. Using a double-barreled pipette attached to a pressure injector, small volumes (2-10 nl) of ATP (0.01-1 mM) and P2 receptor agonists or P2 receptor antagonists in artificial endolymph were introduced into the scala media of the first (basal) and third turns of the guinea pig cochlea, while the EP and CM were monitored. ATP and P2 receptor agonists (5x10(-14)-1x10(-11)cibacron blue. Neither adenosine nor uridine 5'-triphosphate (2x10(-13)-2x10(-11) moles) nor the P2 receptor antagonists on their own had any effect on EP and CM. The ATP effect on the potentials was greater at the third cochlear turn when compared to the first turn. These results provide evidence that in the endolymphatic compartment of the guinea pig, the extracellular ATP effect on cochlear function is likely mediated through an interaction with P2 receptors which assemble as ATP-gated ion channels.

Adenosine Triphosphate↗