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Secretion of endolymph by the isolated frog semicircular canal.

Secretion of endolymph is localized in some structures of the inner ear, namely the stria vascularis in the cochlea and the dark cells in the vestibule and in the lower vertebrate inner ear. In isolated semicircular canal it is possible to study separately the endolymphatic composition in the ampulla, which contains the dark cells, and in its non-ampullar part, which is devoid of these cells. Further, in vitro preparation of the semicircular canal provides access to both faces of the epithelium so that different agents can be applied separately to the apical or to the basolateral membranes of the epithelium. In this structure, the following results were obtained: i) in vitro, the semicircular canal secreted a K-rich, positively polarized fluid; ii) this fluid was secreted only in the ampulla of the semicircular canal; iii) the secretion of endolymph was dependent on basolateral Na+, K(+)-ATPase, inhibited by ouabain, and basolateral Na-K-Cl co-transporter, inhibited by bumetanide; iv) approximately 60% of luminal Na absorption occurred across a luminal Na channel inhibited by amiloride; v) the permeability of the paracellular pathway of the semicircular canal epithelium was 7.10(-7) cm/s. These results indicate that endolymph secretion involves basolateral Na+, K(+)-ATPase and Na-K-Cl co-transporter. An Na channel has been shown at the apical membrane.

Animals↗

Longitudinal endolymph movements and endocochlear potential changes induced by stimulation at infrasonic frequencies.

The inner ear is continually exposed to pressure fluctuations in the infrasonic frequency range (< 20 Hz) from external and internal body sources. The cochlea is generally regarded to be insensitive to such stimulation. The effects of stimulation at infrasonic frequencies (0.1 to 10 Hz) on endocochlear potential (EP) and endolymph movements in the guinea pig cochlea were studied. Stimuli were applied directly to the perilymph of scala tympani or scala vestibuli of the cochlea via a fluid-filled pipette. Stimuli, especially those near 1 Hz, elicited large EP changes which under some conditions exceeded 20 mV in amplitude and were equivalent to a cochlear microphonic (CM) response. Accompanying the electrical responses was a cyclical, longitudinal displacement of the endolymph. The amplitude and phase of the CM varied according to which perilymphatic scala the stimuli were applied to and whether a perforation was made in the opposing perilymphatic scala. Spontaneously occurring middle ear muscle contractions were also found to induce EP deflections and longitudinal endolymph movements comparable to those generated by perilymphatic injections. These findings suggest that cochlear fluid movements induced by pressure fluctuations at infrasonic frequencies could play a role in fluid homeostasis in the normal state and in fluid disturbances in pathological states.

Animals↗

Chemical composition of saccular endolymph and otolith in fish inner ear: lack of spatial uniformity.

Fish otoliths provide a record of age, growth, and environmental influences. In both trout and turbot, spatial chemical investigation of the endolymph surrounding the otolith (sagitta) showed a lack of uniformity. Proteins, PO(3-)(4), and Mg(2+) were significantly more concentrated in the proximal (facing the macula) than distal zone, whereas the opposite was observed for K(+) and total CO(2) (totCO(2)). Na(+) concentration ([Na(+)]) was 20% higher in the proximal zone in trout but not in turbot. Total Ca and Cl(-) contents were uniformly distributed in both species. We propose that the endolymphatic gradients of protein and totCO(2) concentration within the endolymph are involved in the otolithic biocalcification process. Microchemical analyses of otolith sections by wavelength dispersive spectrometry showed a lack of spatial uniformity in the K/Ca and Na/Ca ratios, whereas the Sr/Ca ratio was uniform. There is a clear relationship between endolymph and otolith [K(+)], but the interpretation of the results for [Na(+)] needs further investigation. Thus the lack of uniformity in the otolith composition must be taken into account when investigating otolith microchemistry.

Animals↗

Secretion of endolymph by semicircular canals of the shark.

The semicircular canals of the vestibular labyrinth of the dogfish shark, Squalus acanthias, may serve as a simple in vitro system for the study of the ionic transport mechanisms involved in endolymph formation. Electron microscopy showed that the epithelium was made up of at least three distinct cell types divided into separate regions running the length of the canals. Secretion of endolymph was studied in isolated canals by the split droplet method; when the lumens of the canals were filled with shark Ringer a potassium-rich fluid was secreted into the lumen at a rate of 0.34 microliter.cm-2.min-1. The K concentration of the secreted fluid averaged 112 mM; the calculated rate of K secretion was 2.3 mumol.cm-2.h-1, comparable to recent measurements in mammalian utricle. Fluid secretion was dependent on active transport and was inhibited by ouabain, bumetanide, or methazolamide in the external bathing solution. Fluid secretion was unaffected by the K channel blocker, Ba, in the luminal droplet or by the adenylate cyclase stimulator, forskolin, in the external bathing solution. For electrophysiological analysis, isolated canals were perfused in a chamber designed for voltage/current clamping; an axial wire was inserted into the canal lumen and constant-current pulses were passed to determine tissue resistance. When the luminal fluid was high-K Ringer, transepithelial potential difference was -1.1 mV (lumen negative) and resistance 37 omega.cm2. Dilution and bi-ionic potential measurements showed that the epithelium of the canals exhibited only slight cation selectivity. These results are consistent with a model for endolymph secretion involving cotransport secondary to the ion gradients created by the Na+-K+-ATPase.

Animals↗

Development of monovalent ions in the endolymph in mouse cochlea.

The present study was designed to clarify the chronological developmental process of monovalent ions (Na(+), K(+), Cl(-)) in the endolymph of the mouse in relation to the development of the endocochlear potential (EP). The EP and ionic concentrations were measured simultaneously with the ion-sensitive double-barreled microelectrodes from the scala media of the basal turn. The EP increased abruptly 7 days after birth (DAB) and reached approximately 80 mV 14 DAB. In the earliest postnatal days, the endolymphatic Na(+) concentration was significantly higher than that in adult mice, however, the K(+) and the Cl(-) concentrations were lower. The concentrations of all the monovalent ions in endolymph reached adult levels at 7 DAB when the EP was still under 20 mV. These data strongly suggest the presence of a different mechanism between the production of monovalent ions, especially of high K(+) in the endolymph and that of EP.

Animals↗

Adrenergic and muscarinic control of cochlear endolymph production.

The transduction of sound into nerve impulses in the cochlea is dependent on the stria vascularis. It is a multilayered epithelium, which is part of the epithelial barrier between endolymph and perilymph. The current model designed to explain the generation of the endocochlear potential assumes that the molecular mechanism for the generation of the endocochlear potential is the K(IR)4.1 K+ channel localized in the intermediate cells and that strial marginal cells play an indirect role in the generation of the endocochlear potential. This role is limited to the maintenance of a low K+ concentration in the intrastrial space by absorbing K+ from this space and secreting it into the endolymph. The molecular mechanisms for K+ secretion by strial marginal cells are well established. Strial marginal cells absorb K+ from the intrastrial space via the Na+-K+-ATPase and the Na+2Cl-K+ cotransporter and secrete it across the apical membrane via the IsK/KvLQT1 K+ channel. K+ secretion by strial marginal cells is not only required for the maintenance of the endocochlear potential and to provide the charge carrier for the transduction mechanism, but also to maintain a constant volume of endolymph. Thus, the presence of multiple control mechanisms regulating the rate of K+ secretion is likely. Recent observations suggest that the rate of K+ secretion in strial marginal cells is stimulated by beta1-adrenergic receptors and inhibited by M3 and/or M4 muscarinic receptors.

Cochlea↗

Rates of ion movement from plasma to endolymph in the dogfish.

The rates of movement of Na+, Rb+, Cl- and HCO3- from plasma to endolymph were studied in the elasmobranch fish, Squalus acanthias, by use of the appropriate isotopes. Rb+ was used as a marker for K+. The half-times to equilibrium for Na+, Rb+ and Cl- were about 100 hours; for HCO3- it was 6 hours. The equilibrium ratios, endolymph/plasma, are Na+ 0.87, K+ 26, Cl- 1.37, HCO3- 1.47. Carbonic anhydrase inhibition decreased the rate of HCO3- accumulation, suggesting that the process is actually the formation of endolymphatic HCO3- from plasma or tissue CO2. Increase in plasma pCO2 elevates endolymph HCO3- concentration. The secretory tissue contains carbonic anhydrase and Na-K-ATPase. These and other data suggest that a dominant feature of endolymph chemistry may be HCO3- formation linked in some fashion with K+ transport, through rates catalyzed by these two enzymes.

Acetazolamide↗

Pressure relationship between perilymph and endolymph associated with endolymphatic infusion.

The pressure difference between the perilymph and the endolymph following infusion of artificial endolymph into the endolymphatic space was studied in guinea pigs. Both the perilymphatic and the endolymphatic pressures were measured simultaneously with use of two sets of a servo-nulling system. Endolymphatic infusion caused pressure increases in both the endolymphatic and the perilymphatic spaces, but no measurable pressure difference between them. Soon after the endolymphatic infusion, both pressures returned nearly to the initial levels. We conclude that 1) the endolymphatic and the perilymphatic pressures are very closely related, and 2) if a physiologically significant pressure difference exists between endolymph and perilymph in endolymphatic hydrops, it is below the resolving power of the methods used in this study (+/- 0.1 mm Hg).

Animals↗

Free-floating endolymph particles: a new operative finding during posterior semicircular canal occlusion.

Most clinicians accept cupulolithiasis as the pathophysiological mechanism underlying benign paroxysmal positional vertigo (BPPV.) According to this theory, a cupular deposit induces a gravitational effect on the posterior canal crista. Posterior semicircular canal occlusion is a new operative procedure for treating incapacitating BPPV. It is postulated that canal occlusion abolishes endolymph movement within the canal, effectively fixing the cupula and rendering it unresponsive to both angular and linear acceleration (gravity). During two recent canal occlusions, abundant "free-floating particles" were identified within the posterior canal endolymph. When changing the position of the canal in the earth vertical plane, these free-floating particles would move under the influence of gravity. The hydrodynamic drag of the particles would induce endolymph movement with cupular displacement leading to the typical response. This finding supports an alternate explanation to cupulolithiasis as the pathophysiological mechanism underlying BPPV.

Aged↗

Effect of glycerol on electrochemical composition of endolymph and perilymph in the rat.

Glycerol (2 g/kg body weight), or 0.15 M NaCl for control animals, was administered to rats by i.v. injection. The dose was chosen in order to obtain an osmolarity increase in plasma of about 15 mosm/l 1 h after the glycerol administration, an increase which is similar to that observed in the human glycerol dehydration test. Endolymph and perilymph were sampled from the basal turn of the cochlea; cerebrospinal fluid (CSF) was sampled from cisterna magna. Plasma osmolarity, endocochlear potential, Na and K concentrations in endolymph, perilymph and CSF were determined 1 and 2 h after the glycerol injection. Compared with control animals, glycerol induced an increase in Na and K concentration in perilymph and endolymph, respectively, 1 and 2 h after the glycerol injection. No modification of the endocochlear potential was observed. These results are compatible with an increase in inner ear fluids osmolarity induced by glycerol.

Action Potentials↗

[Contribution to the study of endolymph homeostasis].

The secretory structures of the cochlea including stria vascularis and spiral ligament are responsible for the secretion of endolymph, a fluid characterized by a high potassium concentration [150-180 mM], a low sodium concentration [< 1 mM] and a positive potential [80-100 mV]. This intra-cellular-like fluid fills the endolymphatic compartment and is essential in the transduction process which takes place in the organ of Corti. Yet, the mechanisms which control the homeostasis of this fluid remain largely unknown. To approach this issue we investigated the possibility of a steroid synthesis by the rat cochlea which might modulate the secretion of endolymph. Results show that inner ear expresses mARN encoding some of the enzymes of the steroid pathways. We also investigated the presence of the gastric H+, K(+)-ATPase pump. Results from RT-PCR show that the gastric H+, K(+)-ATPase alpha and beta subunits are expressed in the rat inner ear-lateral wall, organ of Corti and spiral ganglion neurons. The presence of the alpha subunit in inner ear was confirmed by immunoblot. Immunohistochemistry localized this protein in the intermediate cells of the stria vascularis, in the spiral ligament and in spiral ganglion neurons. Along with the vacuolar H(+)-ATPase, the gastric H+, K(+)-ATPase could be involved in the maintenance of H+ equilibrium in endolymph. All these arguments suggest that among the various types of so-called "sensorineural" deafness, some entities including some forms of congenital hearing loss and diuretic-induced ototoxic deafness should be classified as endolymphatic deafness. Such identification seems necessary since these entities result from different pathogenetic mechanisms and might benefit from the development of new therapies.

Animals↗

Immunological analysis of IgG and other protein fractions in endolymph obtained from endolymphatic sac of Meniere patients and a control.

During endolymphatic sac surgery on patients with Meniere's disease, very small amounts (1 microliter) of the intra-sac endolymph were obtained with micro-glass pipets. An immunological assay of microanalysis of the protein by the single radial immunodiffusion (SRID) method was applied to 2 patients, one with bilateral involvement and the other unilateral. Among the protein fractions Alb, IgG, IgM, IgA, C3, C4, transferrin and alpha 2M, the concentrations of IgG were relatively higher, especially in the Meniere patient in the active stage. From 2 other patients and a control deaf patient with temporal bone fracture, the same amounts of endolymph were obtained, accompanied by a tiny specimen of the outer wall of the sac. When a radio-immunological procedure was applied to the endolymph and the homogenized sac, the specific concentration of IgG was higher in the Meniere patients than in the control.

Albumins↗

[Biochemical analysis of the endolymph and perilymph of the trout (Salmo gairdneri) (author's transl)].

Cerebrospinal fluid, perilymph and endolymph of the saccule, the utricle and the semicircular duct of the trout were prepared from frozen inner ears. Blood was collected after the decapitation of the animal. In all samples of each animal the concentrations of sodium, potassium, protein and glucose were determined. The electrolyte values and the total protein content were used to estimate the purity of a sample. The concentration of sodium is high in perilymph and low in endolymph whereas potassium is high in endolymph and low in perilymph. The differences between perilymph and the cerebrospinal fluid as well as the blood serum are significant. A relative independent production of the inner ear fluids can be assumed.

Animals↗

Effect of acetazolamide on cation concentration in the endolymph of the endolymphatic sac.

Acetazolamide (ACTZ), a carbonic anhydrase inhibitor, has been reported to decrease the endolymphatic sac (ES) DC potential (ESP) in the guinea pig. To assess the involvement of cation transport in the ESP change by ACTZ we examined the effect of ACTZ upon the K+ and Na+ activities of the ES endolymph in the guinea pig using ion-sensitive microelectrode. ACTZ (10 mg/kg), a dose that produces the ESP maximum reduction, produced a significant increase in Na+ activity of the ES endolymph with no change in K+ activity. The results suggest that Na+ transport may be directly or indirectly involved in ESP reduction by ACTZ, and that a Na(+)-H+ exchanger may be involved in Na+ influx pathway from endolymph to the ES epithelial cells.

Acetazolamide↗

Evidence for a perilymphatic origin of the endolymph: application to the pathophysiology of Ménière's disease.

The origin of the endolymph was elucidated by kinetic studies of the entry of water and electrolytes into endolymph and perilymph after intravenous administration of radioactive tracers in rats. The compartmental analysis of the data and the comparison of this study with the results of Konishi and associates (Acta Otolaryngol (Stockh) 86, 22-34 and 176-184, 1978), using perilymphatic perfusion of tracers, indicate that perilymph rather than plasma may be considered the precursor of endolymph. Since the cochlear epithelium was found to be freely permeable to water, an alteration of electrolyte transportation across the membranous labyrinth may be involved in the pathophysiology of Ménière's disease. Chloride transport across the cochlear epithelium was investigated using acetazolamide, a specific carbonic anhydrase inhibitor.

Acetazolamide↗

Human endolymphatic duct: possible mechanisms of endolymph outflow.

The ultrastructure of the normal human endolymphatic duct (ED) was observed by transmission electron microscopy. The role of the epithelium, the various regions of the subepithelial space, and vasculature in the resorption of endolymph was morphologically studied in order to generate testable hypotheses of human ED function. These hypothetical mechanisms of endolymph outflow at the level of the ED are a passive transcellular movement of water across the epithelium, driven by an osmotic gradient created by a subepithelial organic matrix; an active transcellular ion exchange with a passive transepithelial outflow of water, which stresses the importance of the dilated lateral intercellular spaces; and an active transcellular vacuolar endolymph outflow, whereby high molecular weight substances are removed by the ED. These mechanisms may be useful in designing experimental studies of the ED and in interpretation of retrospective light microscopic and transmission electron microscopic studies of patients with Meniere's disease.

Adult↗

Austin endolymph dispersement shunt surgery for Menière's disease.

This paper analyses the results of 53 Austin endolymph dispersement shunt procedures as compared to previously reported data on 87 Arenberg inner ear valve implants and 339 cases employing various other surgical techniques. Overall results are similar regardless of the surgical method involved, however findings indicate that capillary endolymph dispersement both affords more class A results, with isolated spectacular hearing gains, and creates a greater chance of completely eliminating dizziness in both definite and adjunctive spells. Furthermore, Austin's endolymph dispersement drain evidenced no adverse effects on inner ear function, unlike Arenberg's inner ear valve, and is herein documented as a safe and effective surgical option for the treatment of Meniere's disease.

Adult↗

Development of endolymph during maturation of the mammalian inner ear. A preliminary report.

The development of the elemental composition in the endolymphatic space was investigated during embryologic and early post natal maturation of the CBA/CBA mouse. At birth the elemental distribution was similar in the endo- and perilymphatic spaces. Mature composition of endolymph was reached 6--8 days post partum. The maturation of endolymph corresponded well in time with the morphological maturation of the stria vascularis.

Age Factors↗