Endolymphatic potassium of the chicken vestibule during embryonic development

The endolymph fills the lumen of the inner ear membranous labyrinth. Its ionic composition is unique in vertebrates as an extracellular fluid for its high-K(+)/low-Na(+) concentration. The endolymph is actively secreted by specialized cells located in the vestibular and cochlear epithelia. We have investigated the early phases of endolymph secretion by measuring the endolymphatic K(+) concentration in the chicken vestibular system during pre-hatching development. Measurements were done by inserting K(+)-selective microelectrodes in chicken embryo ampullae dissected at different developmental stages from embryonic day 9 up to embryonic day 21 (day of hatching). We found that the K(+) concentration is low (<10mM/L) up to embryonic day 11, afterward it increases steeply to reach a plateau level of about 140 mM/L at embryonic day 19--21. We have developed a short-term in vitro model of endolymph secretion by culturing vestibular ampullae dissected from embryonic day 11 chicken embryos for a few days. The preparation reproduced a double compartment system where the luminal K(+) concentration increased along with the days of culturing. This model could be important for (1) investigating the development of cellular mechanisms contributing to endolymph homeostasis and (2) testing compounds that influence those mechanisms

Calyx and dimorphic neurons of mouse Scarpa's ganglion express histamine H3 receptors

<p>Abstract</p> <p>Background</p> <p>Histamine-related drugs are commonly used in the treatment of vertigo and related vestibular disorders. The site of action of these drugs however has not been elucidated yet. Recent works on amphibians showed that histamine H3 receptor antagonists, e.g. betahistine, inhibit the afferent discharge recorded from the vestibular nerve. To assess the expression of H3 histamine receptors in vestibular neurons, we performed mRNA RT-PCR and immunofluorescence experiments in mouse Scarpa's ganglia.</p> <p>Results</p> <p>RT-PCR analysis showed the presence of H3 receptor mRNA in mouse ganglia tissue. H3 protein expression was found in vestibular neurons characterized by large and roundish soma, which labeled for calretinin and calbindin.</p> <p>Conclusion</p> <p>The present results are consistent with calyx and dimorphic, but not bouton, afferent vestibular neurons expressing H3 receptors. This study provides a molecular substrate for the effects of histamine-related antivertigo drugs acting on (or binding to) H3 receptors, and suggest a potential target for the treatment of vestibular disorders of peripheral origin.</p

Signal Transmission by Auditory and Vestibular Hair Cells

We interact with the world around us by sensing a vast array of inputs and translating them into signals that can be interpreted by the brain. We have evolved many sensory receptors, each uniquely specialised to detect diverse stimuli. The hair cells are sensory receptors, initially developed to provide a sense of body position and movement, but later adapted to sense minute pressure waves in the environment that are perceived as sounds. As such, hair cells bestow a sense of hearing and balance, which are major advantages for survival. Mammals have four different types of hair cell, two of which are dedicated to hearing, the inner and outer hair cells, and the other two to balance, the type-I and type-II hair cells. While all hair cells employ common mechanisms to detect and relay signals from sound or motion, they also have unique attributes that specialise them for a specific functional role. In this chapter we describe the process of signal transmission in mammalian auditory and vestibular hair cells. Since mammalian hair cells do not regenerate, their loss results in permanent auditory or vestibular deficit. Efforts to regenerate or repair malfunctioning hair cells have recently intensified, mainly through gene, stem-cell and molecular therapy

Aquaporin-6 Expression in the Cochlear Sensory Epithelium Is Downregulated by Salicylates

We characterize the expression pattern of aquaporin-6 in the mouse inner ear by RT-PCR and immunohistochemistry. Our data show that in the inner ear aquaporin-6 is expressed, in both vestibular and acoustic sensory epithelia, by the supporting cells directly contacting hair cells. In particular, in the Organ of Corti, expression was strongest in Deiters' cells, which provide both a mechanical link between outer hair cells (OHCs) and the Organ of Corti, and an entry point for ion recycle pathways. Since aquaporin-6 is permeable to both water and anions, these results suggest its possible involvement in regulating OHC motility, directly through modulation of water and chloride flow or by changing mechanical compliance in Deiters' cells. In further support of this role, treating mice with salicylates, which impair OHC electromotility, dramatically reduced aquaporin-6 expression in the inner ear epithelia but not in control tissues, suggesting a role for this protein in modulating OHCs' responses

miR-96 regulates the progression of differentiation in mammalian cochlear inner and outer hair cells

MicroRNAs (miRNAs) are small noncoding RNAs able to regulate a broad range of protein-coding genes involved in many biological processes. miR-96 is a sensory organ-specific miRNA expressed in the mammalian cochlea during development. Mutations in miR-96 cause nonsyndromic progressive hearing loss in humans and mice. The mouse mutant diminuendo has a single base change in the seed region of the Mir96 gene leading to widespread changes in the expression of many genes. We have used this mutant to explore the role of miR-96 in the maturation of the auditory organ. We found that the physiological development of mutant sensory hair cells is arrested at around the day of birth, before their biophysical differentiation into inner and outer hair cells. Moreover, maturation of the hair cell stereocilia bundle and remodelling of auditory nerve connections within the cochlea fail to occur in miR-96 mutants. We conclude that miR-96 regulates the progression of the physiological and morphological differentiation of cochlear hair cells and, as such, coordinates one of the most distinctive functional refinements of the mammalian auditory system

Elementary properties of CaV1.3 Ca2+ channels expressed in mouse cochlear inner hair cells

Mammalian cochlear inner hair cells (IHCs) are specialized to process developmental signals during immature stages and sound stimuli in adult animals. These signals are conveyed onto auditory afferent nerve fibres. Neurotransmitter release at IHC ribbon synapses is controlled by L-type CaV1.3 Ca2+ channels, the biophysics of which are still unknown in native mammalian cells. We have investigated the localization and elementary properties of Ca2+ channels in immature mouse IHCs under near-physiological recording conditions. CaV1.3 Ca2+ channels at the cell pre-synaptic site co-localize with about half of the total number of ribbons present in immature IHCs. These channels activated at about −70 mV, showed a relatively short first latency and weak inactivation, which would allow IHCs to generate and accurately encode spontaneous Ca2+ action potential activity characteristic of these immature cells. The CaV1.3 Ca2+ channels showed a very low open probability (about 0.15 at −20 mV: near the peak of an action potential). Comparison of elementary and macroscopic Ca2+ currents indicated that very few Ca2+ channels are associated with each docked vesicle at IHC ribbon synapses. Finally, we found that the open probability of Ca2+ channels, but not their opening time, was voltage dependent. This finding provides a possible correlation between presynaptic Ca2+ channel properties and the characteristic frequency/amplitude of EPSCs in auditory afferent fibres

Fine Tuning of Ca(V)1.3 Ca2+ Channel Properties in Adult Inner Hair Cells Positioned in the Most Sensitive Region of the Gerbil Cochlea

Hearing relies on faithful signal transmission by cochlear inner hair cells (IHCs) onto auditory fibres over a wide frequency and intensity range. Exocytosis at IHC ribbon synapses is triggered by Ca2+ inflow through CaV1.3 (L-type) Ca2+ channels. We investigated the macroscopic (whole-cell) and elementary (cell-attached) properties of Ca2+ currents in IHCs positioned at the middle turn (frequency ,2 kHz) of the adult gerbil cochlea, which is their most sensitive hearing region. Using near physiological recordings conditions (body temperature and a Na+ based extracellular solution), we found that the macroscopic Ca2+ current activates and deactivates very rapidly (time constant below 1 ms) and inactivates slowly and only partially. Single-channel recordings showed an elementary conductance of 15 pS, a sub-ms latency to first opening, and a very low steady-state open probability (Po: 0.024 in response to 500-ms depolarizing steps at ,218 mV). The value of Po was significantly larger (0.06) in the first 40 ms of membrane depolarization, which corresponds to the time when most Ca2+ channel openings occurred clustered in bursts (mean burst duration: 19 ms). Both the Po and the mean burst duration were smaller than those previously reported in high-frequency basal IHCs. Finally, we found that middle turn IHCs are likely to express about 4 times more Ca2+ channels per ribbon than basal cells. We propose that middle-turn IHCs finely-tune CaV1.3 Ca2+ channel gating in order to provide reliable information upon timing and intensity of lower-frequency sounds

Hair cells

Hair cells are the sensory receptors in the inner ear that detect sound and head motion to begin the processes of hearing and balance control. The defining feature of hair cells is the hair bundle, the transduction organelle protruding from their apical surface composed of ordered arrays of stereocilia. Mechanical deflection of the hair bundle, normally induced by physiological stimuli, increases the open probability of mechanically gated cation channels located at the tip of stereocilia. The resulting depolarizing inward current generates a receptor potential. The information encoded in this electrical response is transmitted to the auditory or vestibular afferent nerve fibres via the Ca2+-induced release of neurotransmitter from the hair cell’s basal pole. In this way sensory information is relayed to the brain enabling us to perceive sound and maintain balance. In mammals, hair cell loss causes irreversible balance and hearing impairment because these sensory cells show very little or no regenerative ability

Hair Cells

The auditory and the vestibular systems use hair cells (HCs) as their sensory receptors. HCs are neu roepithelial cells characterised by the presence of a bundle of microvilli-like structures that protrude from their apical surface, called stereocilia. The displacement of stereocilia, which is caused by acoustic stimuli in the cochlea or head movement in the vestibule, is converted into a depolarising inward current by mechanoelectrical transducer (MET) channels located at their tip. The depolarisa tion of HCs opens voltage-dependent Ca2+ channels at their basolateral synaptic active zones, which are functionally coupled to glutamate-containing vesicles at specialised ribbon synapses. There is also evidence for a nonquantal synaptic transmis sion at the vestibular HCs, likely involving direct postsynaptic depolarisation by K+ exiting the cells. In mammals, HC loss causes irreversible balance and hearing impairment because these cells do not regenerate

Elementary properties of Kir2.1, a strong inwardly rectifying K(+) channel expressed by pigeon vestibular type II hair cells

By using the patch-clamp technique in the cell-attached configuration, we have investigated the single-channel properties of an inward rectifier potassium channel (Kir) expressed by pigeon vestibular type II hair cells in situ. In high-K(+) external solution with 2 mM Mg(2+), Kir inward current showed openings to at least four amplitude levels. The two most frequent open states (L2 and L3) had a mean slope conductance of 13 and 28 pS, respectively. L1 (7 pS) and L4 (36 pS) together accounted for less than 6% of the conductive state. Closed time distributions were fitted well using four exponential functions, of which the slowest time constant (tau(C4)) was clearly voltage-dependent. Open time distributions were fitted well with two or three exponential functions depending on voltage. The mean open probability (P(O)) decreased with hyperpolarization (0.13 at -50 mV and 0.03 at -120 mV). During pulse-voltage protocols, the Kir current-decay process (inactivation) accelerated and increased in extent with hyperpolarization. This phenomenon was associated with a progressive increase of the relative importance of tau(C4). Kir inactivation almost disappeared when Mg(2+) was omitted from the pipette solution. At the same time, P(O) increased at all membrane voltages and the relative importance of L4 increased to a mean value of 47%. The relative importance of tau(C4) decreased for all open states, while L4 only showed a significantly longer open time constant. The present work provides the first detailed quantitative description of the elementary properties of the Kir inward rectifier in pigeon vestibular type II hair cells and specifically describes the Kir gating properties and the molecule's sensitivity to extracellular Mg(2+) for all subconductance levels. The present results are consistent with the Kir2.1 protein sustaining a strong inwardly rectifying K(+) current in native hair cells, characterized by rapid activation time course and slow partial inactivation. The longest closed state (tau(C4)) appears as the main parameter involved in time- and Mg(2+)-dependent decay. Finally, in contrast to Kir2.1 results described so far for mammalian cells, external Mg(2+) had no effect on channel conductance