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At least 19 recordsLinked to original sources
PubMed1991

Motile responses of isolated guinea pig vestibular hair cells.

Vestibular hair cells were isolated from the guinea pig vestibule by a micromechanical non-enzymatic procedure. Perfusion with 125 mM K+ solution induced irreversible slow shortening of the necks in 42.8% of the hair cells tested. Mechanical stimulation, creating a displacement of the hair bundle towards the kinocilium, induced either irreversible coiling or tilting of the neck of the cells, or reversible fast tilting of the cuticular plate (44.5% of tested cells). The response to the Ca2+ antagonist, Flunarizine, suggested that these movements were calcium-dependent. We propose several explanations of the physiological role of these mechanisms and discuss the possibility that fast tilting of the cuticular plate is a physiological movement involving the hair cells at the periphery of the vestibular receptors. The regulation of the vestibular message at the apex of type I hair cells is also considered.

Animals↗
PubMed1992

Electrically evoked motile responses of mammalian type I vestibular hair cells.

Vestibular hair cells, type I and II, with membrane potentials around -64 mV were prepared from guinea pig ampullar cristae and maculae. In type I cells, current injection, application of voltage steps during membrane patch-clamping, or extracellular alternating current (ac) fields evoked fast length changes of 50 nm to 500 nm of the cell "neck." Mechanical responses were determined by computerized video techniques with contrast-enhanced digital image subtraction (DIS) and interpeak pixel counts (IPPC) or by double photodiode measurements. These techniques allowed spatial resolutions of 300 nm, 120 nm, and 50 nm, respectively. In contrast to measurements of high-frequency movements of auditory outer hair cells (OHCs), the mechanical responses of type I VHCs were restricted to low frequencies below 85 Hz. In addition to recently reported slow motility of VHCs, the present results suggest that fast mechanical VHC responses could significantly influence macular and cupular mechanics. Isometric and isotonic variants are discussed. The observed frequency maxima gap between VHCs and OHCs is suggested to contribute to a clear separation of the auditory and the vestibular sensory modality.

Animals↗
PubMed1994

Motile responses of isolated vestibular hair cells.

Vestibular sensory cells were isolated from the utricular macula or crista ampullaris of the guinea pig by enzymatic and mechanical dissociation. Isolated vestibular hair cells, especially type I hair cells, showed an active motile capacity. After exposure to a medium containing high concentration of potassium, or to a hypoosmotic medium, the type I hair cells showed tilting of their hair bundle to about 15 degrees. Given the tight and dense structure of the vestibular epithelium, the changes in shape of the isolated vestibular hair cells may in vivo lead to an influence of the mechano-sensitive stereocilia and modulate stiffness and compliance of the receptor structure as a whole including its cupular or macular relationship. This active mechanical events could be closely related to an active adaptation process.

Animals↗
PubMed1996

The delayed rectifier, IKI, is the major conductance in type I vestibular hair cells across vestibular end organs.

Hair cells were dissociated from the semicircular canal, utricle, lagena and saccule of white king pigeons. Type I hair cells were identified morphologically based on the ratios of neck width to cuticular plate width (NPR < 0.72) as well as neck width to cell body width (NBR < 0.64). The perforated patch variant of the whole-cell recording technique was used to measure electrical properties from type I hair cells. In voltage-clamp, the membrane properties of all identified type I cells were dominated by a predominantly outward potassium current, previously characterized in semicircular canal as IKI. Zero-current potential, activation, deactivation, slope conductance, pharmacologic and steady-state properties of the complex currents were not statistically different between type I hair cells of different vestibular end organs. The voltage dependence causes a significant proportion of this conductance to be active about the cell's zero-current potential. The first report of the whole-cell activation kinetics of the conductance is presented, showing a voltage dependence that could be best fit by an equation for a single exponential. Results presented here are the first data from pigeon dissociated type I hair cells from utricle, saccule and lagena suggesting that the basolateral conductances of a morphologically identified population of type I hair cells are conserved between functionally different vestibular end organs; the major conductance being a delayed rectifier characterized previously in semicircular canal hair cells as IKI.

4-Aminopyridine↗
PubMed2005

Effects of cinnarizine on calcium and pressure-dependent potassium currents in guinea pig vestibular hair cells.

In vestibular hair cells, K+ currents induced by rises in hydrostatic pressure have recently been demonstrated. These currents are inhibited by charybdotoxin, a blocker of Ca2+-dependent K+ channels. On the other hand, cinnarizine is a blocker of voltage-gated Ca2+ currents in hair cells and is used as a drug in conditions with vestibular vertigo. Our aim was to test in patch-clamp experiments (conventional whole-cell mode) whether cinnarizine, by reducing Ca2+ influx, inhibited Ca2+ and pressure-sensitive K+ currents in vestibular type-II hair cells of guinea pigs. A quantitatively similar inhibition of K+ currents was evoked by extracellular Ca2+ removal, cinnarizine (0.5 microM), and the L-type Ca2+ channel blocker nifedipine (3 microM). Cinnarizine abrogated increases of K+ currents induced by increases in the hydrostatic pressure (from 0.2 to 0.5 cm H2O). At a higher concentration (1 microM), cinnarizine elicited K+ current inhibitions larger than those elicited by Ca2+ removal. Moreover, it reduced K+ currents in the absence of Ca2+, in contrast to nifedipine. However, charybdotoxin abolished these effects of cinnarizine. We thus conclude that cinnarizine inhibits, by two mechanisms, pressure-induced currents that are sensitive to charybdotoxin and Ca2+. It reduces Ca2+ influx and exerts a Ca2+-independent inhibition, with a lower IC50 than that required for Ca2+ channel blockade. These two actions may importantly contribute to its therapeutic effects.

Animals↗
PubMed1993

Voltage dependent reversible movements of the apex in isolated guinea pig vestibular hair cells.

Type I vestibular hair cells isolated from guinea pig were placed in the whole cell clamp configuration, and electrically stimulated by depolarizing voltage pulses. The voltage dependent reversible movements of the cell apex affected the length of the cell neck, the position of the cuticular plate, and the tilting and bending of the stereocilia. The cell neck shortened when the membrane was depolarized by 10 mV while cuticular plate and the stereocilia tilting did not begin until 20 mV. The shortening was 0.5 to 1 micron, and the cuticular plate tilting was up to 15 degrees for depolarization amplitudes of 20-40 mV. These movements were reversed within a few seconds. More complex, larger movements were induced by stronger depolarizations. The cuticular plate tilting and the hair bundle bending were always in the opposite direction to the kinocilium position. The small reversible movements of the mammalian type I vestibular hair cells are discussed in terms of mechanical adaptation processes and morphological features. It is suggested that such active movements of the vestibular hair cells occur in vivo.

Animals↗
PubMed1996

[Motility and cytoskeleton of isolated vestibular hair cells of the guinea pig].

Vestibular hair cells were isolated from the vestibular endorgan of the guinea pig by enzymatic and mechanical dissociation. The isolated cells were divided into three types: flask-shaped type I cells, rod-shaped type II cells and round supporting cells. The cilia of type I cells in the crista were longer than those in the corresponding cell type in the macula. After exposure to a medium containing a high concentration of potassium, to a hypoosmotic (280mOsm) medium or to medium containing ATP, the type I cells showed tilting of the neck portion accompanied by tilting of the hair bundle. Isolated living type I vestibular cells are capable of self-movement. Actin and the other proteins associated with the cytoskeleton are thought to be related to vestibular hair cell motility. The location of F-actin in the isolated vestibular hair cell was studied by using FITC-labeled phalloidin. In hair cells fixed by paraformaldehyde, actin filaments found to be located in the hair bundle and cuticular plate. In frozen-fixed cells perfused by artificial perilymph, the filaments were heavily located in the hair bundle and cuticular plate as well as throughout the cytoplasm. In frozen-fixed cells perfused with artificial endolymph containing a high concentration of potassium, staining of the hair bundle and cuticular plate was weaker. These changes in the actin staining pattern may be closely related to the mechanism of self-movement. Given the tight and dense structure of the vestibular epithelium, the shape changes of the isolated vestibular hair cells may in vivo lead to modulate stiffness of the apical portion of type I cells, and compliance of the receptor structure as a whole including its cupular or macular relationship. These active mechanical events could be closely related to an active adaptation process.

Animals↗
PubMed2005

Muscarinic ACh receptor activation causes transmitter release from isolated frog vestibular hair cells.

In the frog, vestibular efferent fibers innervate only type-II vestibular hair cells. Through this direct contact with hair cells, efferent neurons are capable of modifying transmitter release from hair cells onto primary vestibular afferents. The major efferent transmitter, acetylcholine (ACh), is known to produce distinct pharmacological actions involving several ACh receptors. Previous studies have implicated the presence of muscarinic ACh receptors on vestibular hair cells, although, surprisingly, a muscarinic-mediated electrical response has not been demonstrated in solitary vestibular hair cells. This study demonstrates that muscarinic receptors can evoke transmitter release from vestibular hair cells. Detection of this release was obtained through patch-clamp recordings from catfish cone horizontal cells, serving as glutamate detectors after pairing them with isolated frog semicircular canal hair cells in a two-cell preparation. Although horizontal cells alone failed to respond to carbachol, application of 20 microM carbachol to the two-cell preparation resulted in a horizontal cell response that could be mimicked by exogenous application of glutamate. All of the horizontal cells in the two-cell preparation responded to 20 microM CCh. Furthermore, this presumed transmitter release persisted in the presence of d-tubocurarine at concentrations that block all known hair cell nicotinic ACh receptors. The effect on the detector cell, imparted by the carbachol application to the hair cell-horizontal cell preparation, was blocked both by 2-amino-5-phosphonopentanoic acid, a selective N-methyl-D-aspartate antagonist, and the muscarinic antagonist, atropine. Thus vestibular hair cells from the frog semicircular canal can be stimulated to release transmitter by activating their muscarinic receptors.

Action Potentials↗
PubMed1995

Atypical innervation pattern of human vestibular hair cells.

Human vestibular sensory epithelia of macula utriculi were examined in 3 cases with acoustic neurinoma by intermediate voltage electron microscope. The innervation pattern of vestibular hair cells was studied by means of computer aided three-dimensional reconstruction technique. The sensory epithelia were fairly well preserved. Most of type I and all of type II hair cells appeared normal. However, some type I hair cells were incompletely surrounded by nerve calyces and received direct contact from the efferent nerve endings. These type I hair cells were also innervated by a few neighboring afferent nerve calyces. These atypical type I hair cells constituted 5-8% of the total number of hair cells.

Culture Techniques↗
PubMed2005

Neural connections between embryonic stem cell-derived neurons and vestibular hair cells in vitro.

This study aimed to examine the potential of embryonic stem cell (ESC)-derived neural progenitors for restoration of the neural network in the peripheral vestibular system. Mouse ESC-derived neural progenitors were co-cultured with explants of vestibular sensory epithelia from neonatal mice. Histological analyses demonstrated that ESC-derived neurons substantially elongated their neurites towards vestibular hair cells, and attached to hair cells at the regions corresponding to the location of nerve endings in normal vestibular epithelia. Immunoreactivity for synaptophysin, a marker for synaptic vesicles, was present only in the cytoplasm of hair cells in sensory epithelia cultured alone, while the nerve endings of ESC-derived neurons attached to hair cells exhibited intense immunoreactivity for synaptophysin and some hair cells were moderately reactive in co-cultured specimens. The pattern of synaptophysin expression in co-cultured specimens was very similar to that observed in developing sensory epithelia, in which synaptic connections between hair cells and nerve endings are actively formed. These findings indicate that ESC-derived neurons have the potential to restore neural connections in the peripheral vestibular system.

Age Factors↗
PubMedDate not supplied

Cytoskeletal basis for contractility of outer hair cells in the normal adult human organ of Corti: comparisons with vestibular hair cells.

The present study is the first consecutive analysis of the adult normal human organ of Corti and vestibular hair cells with regard to the expression of F-actin, actin-associated proteins (alpha-actinin, alpha- and beta-spectrins, vinculin and tropomyosin), beta-tubulin and the calcium-binding protein synaptophysin. The expression of these cytoskeletal and their associated proteins in man is largely similar to, although not identical with, that previously described for several other mammalian species. However, a few very unusual staining patterns were found. In several long outer hair cells a rod of F-actin extended from the infracuticular area to the cell nucleus. Fluorescence for tropomyosin occurred both in the cuticular plates of the outer and inner hair cells, and in the area of close apposition between the base of the outer hair cell and the apical part of Deiter's cell. In contrast, the vestibular hair cells showed immunoreactivity for tropomyosin only in the cuticular plates.

Actinin↗
PubMed2004

Signaling pathway for apoptosis of vestibular hair cells of mice due to aminoglycosides.

Previous studies on regeneration of mammalian vestibular hair cells have indicated the potential for self-repair of damaged hair cells. The rescue of damaged hair cells from cell death may therefore increase regenerated hair cells in affected vestibular epithelia. The role of apoptosis in the degradation of vestibular hair cells following aminoglycoside treatment has been elucidated. To seek a method of protecting vestibular hair cells from aminoglycoside toxicity, we examined the apoptosis signaling pathway of vestibular hair cells due to aminoglycoside toxicity. Induction of apoptosis in hair cells of mouse ampullar cristae damaged by local application of neomycin was evaluated by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method and transmission electron microscopy (TEM). Immunohistochemistry for apoptosis-related proteins was employed to determine the signaling pathway of apoptosis of hair cells. The occurrence of apoptosis in hair cells was demonstrated by TUNEL staining and TEM. In apoptotic hair cells, activation of caspase-3 and -9, and redistribution of cytochrome c was identified, while there was no expression of activated caspase-8 or apoptosis-inducing factor. In conclusion, these findings indicate that the mitochondria-mediated pathway of apoptosis may play a role in inducing the apoptosis of vestibular hair cells due to aminoglycoside toxicity. Stabilization of the mitochondrial membrane may therefore rescue vestibular hair cells from apoptosis, leading to an increase in self-repaired hair cells in affected vestibular epithelia.

Animals↗
PubMedDate not supplied

Mechanoelectrical and voltage-gated ion channels in mammalian vestibular hair cells.

Mammalian vestibular afferents respond robustly to head movements at low frequencies and provide input to reflexes that control eye, head and body position. Vestibular organs have distinctive regions and hair cells: Type II cells receive bouton afferent endings and type I cells receive large calyx afferent endings. In the rodent utricle, type II cells are broadly tuned to frequencies between 10 and 30 Hz. Other recent data suggest that otolith organs function in this frequency range, which is higher than previously imagined. Some of the tuning derives from adaptation of the transducer current, which is best fitted with a double exponential decay with time constants of approximately 4 and 40 ms. Further tuning is provided by basolateral conductances, principally outwardly rectifying, voltage-gated K+ conductances. The kinetics of the K+ currents tend to vary with location in the sensory epithelium and therefore may contribute to regional variation in afferent physiology. Type I hair cells have a large, negatively activating K+ conductance, g(K,L), that confers a very low input resistance and therefore attenuates the receptor potential. This may reduce nonlinearity in the receptor potential, a possibly useful feature for the motor reflexes served by the vestibular system. On the other hand, the small receptor potentials together with unusually negative resting potentials are hard to reconcile with calcium-mediated quantal transmission. This problem may be overcome by factors that inhibit g(K,L)'s activation at resting potential. Also, the calyx may support nonquantal transmission.

Animals↗
PubMed2004

Differences between the negatively activating potassium conductances of Mammalian cochlear and vestibular hair cells.

Cochlear and type I vestibular hair cells of mammals express negatively activating potassium (K(+)) conductances, called g(K,n) and g(K,L) respectively, which are important in setting the hair cells' resting potentials and input conductances. It has been suggested that the channels underlying both conductances include KCNQ4 subunits from the KCNQ family of K(+) channels. In whole-cell recordings from rat hair cells, we found substantial differences between g(K,n) and g(K,L) in voltage dependence, kinetics, ionic permeability, and stability during whole-cell recording. Relative to g(K,L), g(K,n) had a significantly broader and more negative voltage range of activation and activated with less delay and faster principal time constants over the negative part of the activation range. Deactivation of g(K,n) had an unusual sigmoidal time course, while g(K,L) deactivated with a double-exponential decay. g(K,L), but not g(K,n), had appreciable permeability to Cs(+). Unlike g(K,L), g(K,n)'s properties did not change ("wash out") during the replacement of cytoplasmic solution with pipette solution during ruptured-patch recordings. These differences in the functional expression of g(K,n) and g(K,L) channels suggest that there are substantial differences in their molecular structure as well.

Animals↗
PubMed1997

[Motility of the vestibular hair cell of the guinea pig and bull frog].

The motile response of the isolated vestibular hair cell induced by a neurotransmitter was studied. After application of both physostigmine and acetylcholine (Ach) as well as glutamic acid, shortening or tilting of the neck of the guinea pig hair cell was observed. These findings suggest that the effect of a neurotransmitter in the neck region as well as the efferent neuron is involved in the motile response. The location of F-action in isolated vestibular hair cells was investigated by using FITC-labeled phalloidin. In freeze-fixed vestibular hair cells, marked labeling was noted in the hair bundle, cuticular plate and throughout the cytoplasm. After application of both physostigmine and Ach, the labeling in the cuticular plate and the cytoplasm became more intense than that in the hair bundle. Alteration of this phalloidin-labeling pattern suggests that actin could play an important role in the self movement of vestibular sensory cells. The shape of the bull frog hair cell also changed after application of Ach. At the same time, spontaneous discharge and the time constant of the posterior semicircular canal nerve activity decreased. These results suggest that an adaptation mechanism induced by change in the cell shape and membrane potential inhibits the activity of the afferent neuron. Furthermore, these active events could be closely related to the active regulation of vestibular hair cell transmission.

Acetylcholine↗
PubMed2002

Early development and degeneration of vestibular hair cells in bronx waltzer mutant mice.

In bronx waltzer mouse mutants, inner hair cells die at an early stage in their development, from around 17.5 days of gestation onwards. In contrast, outer hair cells appear to develop normally. Vestibular hair cells also degenerate, but the earliest signs of vestibular abnormalities have not yet been described. We looked at prenatal and early postnatal stages of vestibular development by scanning electron microscopy in the mutants, and established that vestibular hair cells (types I and II) never reach beyond the middle stages of differentiation (at least up to P2) and instead show signs of degeneration. Thus, it appears that the bronx waltzer gene product is required for the continued survival and differentiation of inner and vestibular hair cells past a set point in their development.

Animals↗
PubMed2001

Growth factor treatment enhances vestibular hair cell renewal and results in improved vestibular function.

The vestibules of adult guinea pigs were lesioned with gentamicin and then treated with perilymphatic infusion of either of two growth factor mixtures (i.e., GF I or GF II). GF I contained transforming growth factor alpha (TGFalpha), insulin-like growth factor type one (IGF-1), and retinoic acid (RA), whereas GF II contained those three factors and brain-derived neurotrophic factor. Treatment with GF I significantly enhanced vestibular hair cell renewal in ototoxin-damaged utricles and the maturation of stereociliary bundle morphology. The addition of brain-derived neurotrophic factor to the GF II infusion mixture resulted in the return of type 1 vestibular hair cells in ototoxin-damaged cristae, and improved vestibular function. These results suggest that growth factor therapy may be an effective treatment for balance disorders that are the result of hair cell dysfunction and/or loss.

Animals↗
PubMed1998

Time-dependent response of vestibular hair cells of guinea pigs following high-dose applications of streptomycin.

Apoptosis has been reported to occur in vestibular hair cells following aminoglycoside treatment and is suggested to play a predominant role in deletion of affected hair cells. However, the type of cell death occurring during an acute phase of vestibular damage following high-dose application of streptomycin has not yet been determined. Hence, in this study we examined the cell death mode of vestibular hair cells during the acute phase. The numbers of hair cell nuclei stained by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) method and residual hair cells were quantitatively analysed. Hoechst nuclear staining was used for analysis of the nuclear morphology of affected hair cells. TUNEL staining of hair cell nuclei and lost hair cells began to appear 6 h after streptomycin treatment and increased with more exposure time. Apoptotic nuclear features could also be found from 6 h after streptomycin treatment. These findings support the thesis that apoptosis is a predominant cell death mode in degeneration of vestibular hair cells due to streptomycin ototoxicity.

Animals↗