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R A Eatock

Publications and source records attributed to R A Eatock.

At least 19 recordsLinked to original sources
PubMed2000

Essential role of BETA2/NeuroD1 in development of the vestibular and auditory systems.

BETA2/NeuroD1 is a bHLH transcription factor that is expressed during development in the mammalian pancreas and in many locations in the central and peripheral nervous systems. During inner ear ontogenesis, it is present in both sensory ganglion neurons and sensory epithelia. Although studies have shown that BETA2/NeuroD1 is important in the development of the hippocampal dentate gyrus and the cerebellum, its functions in the peripheral nervous system and in particular in the inner ear are unclear. Mice carrying a BETA2/NeuroD1 null mutation exhibit behavioral abnormalities suggestive of an inner ear defect, including lack of responsiveness to sound, hyperactivity, head tilting, and circling. Here we show that these defects can be explained by a severe reduction of sensory neurons in the cochlear-vestibular ganglion (CVG). A developmental study of CVG formation in the null demonstrates that BETA2/NeuroD1 does not play a primary role in the proliferation of neuroblast precursors or in their decision to become neuroblasts. Instead, the reduction in CVG neuron number is caused by a combination both of delayed or defective delamination of CVG neuroblast precursors from the otic vesicle epithelium and of enhanced apoptosis both in the otic epithelium and among those neurons that do delaminate to form the CVG. There are also defects in differentiation and patterning of the cochlear duct and sensory epithelium and loss of the dorsal cochlear nucleus. BETA2/NeuroD1 is, thus, the first gene to be shown to regulate neuronal and sensory cell development in both the cochlear and vestibular systems.

Acoustic Stimulation↗
PubMed2000

Adaptation in hair cells.

Hair cells adapt to sustained deflections of the hair bundle via Ca(2+)-dependent negative feedback on the open probability of the mechanosensitive transduction channels. A model posits that adaptation relieves the input to the transduction channels--force applied by elastic tip links between stereocilia--by repositioning the insertions of the links in the stereocilium. The tip link insertion and transduction channel are dragged by myosins moving on the stereocilium's actin core. This model accounts for many aspects of adaptation in hair cells of the frog saccule, where adaptation time constants are tens of milliseconds. Adaptation in hair cells of the turtle cochlea is much faster, possibly reflecting a more direct mechanism such as Ca2+ binding to the transduction channel. Adaptation mechanisms attenuate the transduction current at low frequencies and may be tuned to different corner frequencies according to the stimulus demands of the inner ear organ. Other sites of adaptation in the inner ear include accessory structures, voltage-dependent properties of hair cells, and afferent transmitter release. A remaining challenge is to understand how these processes work together to shape the output of the inner ear to natural stimuli.

Acoustic Stimulation↗
PubMed2000

Major potassium conductance in type I hair cells from rat semicircular canals: characterization and modulation by nitric oxide.

Mammalian vestibular organs have two types of hair cell, type I and type II, which differ morphologically and electrophysiologically. Type I hair cells alone express an outwardly rectifying current, I(K, L), which activates at relatively negative voltages. We used whole cell and patch configurations to study I(K,L) in hair cells isolated from the sensory epithelia of rat semicircular canals. I(K,L) was potassium selective, blocked by 4-aminopyridine, and permeable to internal cesium. It activated with sigmoidal kinetics and was half-maximally activated at -74.5 +/- 1.6 mV (n = 35; range -91 to -50 mV). It was a very large conductance (91 +/- 8 nS at -37 mV; 35 nS/pF for a cell of average size). Patch recordings from type I cells revealed a candidate ion channel with a conductance of 20-30 pS. Because I(K,L) was activated at the resting potential, the cells had low input resistances (R(m)): median 25 MOmega at -67 mV versus 1.3 GOmega for type II cells. Consequently, injected currents comparable to large transduction currents (300 pA) evoked small (</=10 mV) voltage responses. The cells' small voltage responses and negative resting potentials (V(R) = -81.3 +/- 0.2 mV, n = 144) pose a problem for afferent neurotransmission: how does the receptor potential depolarize the cell into the activation range of Ca(2+) channels (positive to -60 mV) that mediate transmitter release? One possibility, suggested by spontaneous positive shifts in the activation range of I(K,L) during whole cell recording, is that the activation range might be modulated in vivo. Any factor that reduces the number of I(K,L) channels open at V(R) will increase R(m) and depolarize V(R). Nitric oxide (NO) is an ion channel modulator that is present in vestibular epithelia. Four different NO donors, applied externally, inhibited the I(K,L) conductance at -67 mV, with mean effects ranging from 33 to 76%. The NO donor sodium nitroprusside inhibited channel activity in patches when they were cell-attached but not excised, suggesting an intracellular cascade. Consistent with an NO-cGMP cascade, 8-bromo-cGMP also inhibited whole cell I(K,L). Ca(2+)-dependent NO synthase is reported to be in hair cells and nerve terminals in the vestibular epithelium. Excitatory input to vestibular organs may lead, through Ca(2+) influx, to NO production and inhibition of I(K,L). The resulting increase in R(m) would augment the receptor potential, a form of positive feedback.

4-Aminopyridine↗
PubMed2000

Harvesting human hair cells.

The sensory hair cells of the inner ear are responsible for converting balance and hearing stimuli into electrical signals. Until recently, all previous studies of hair cell physiology had been performed on tissue obtained from non-mammals and rodents. In primates, hair cells are difficult to access, because they rest within the densest structure of the body, the otic capsule of the temporal bone. In this report, we describe a technique that we have used in physiological studies to harvest living human hair cells. We collected vestibular and cochlear tissue specimens from adult humans undergoing translabyrinthine and transotic surgical approaches for resection of lateral skull base tumors. Viable hair cells were identified and visualized with light microscopy. The ability to study normal hair cells from humans may further the study of normal and pathological human sensation, hair cell regeneration, and genetic causes of balance and hearing disorders.

Adult↗
PubMed1999

Math1: an essential gene for the generation of inner ear hair cells.

The mammalian inner ear contains the cochlea and vestibular organs, which are responsible for hearing and balance, respectively. The epithelia of these sensory organs contain hair cells that function as mechanoreceptors to transduce sound and head motion. The molecular mechanisms underlying hair cell development and differentiation are poorly understood. Math1, a mouse homolog of the Drosophila proneural gene atonal, is expressed in inner ear sensory epithelia. Embryonic Math1-null mice failed to generate cochlear and vestibular hair cells. This gene is thus required for the genesis of hair cells.

Animals↗
PubMed1999

Stimulus processing by type II hair cells in the mouse utricle.

In type II and neonatal hair cells in the mouse utricle, the receptor potentials evoked by low-frequency sinusoidal deflections of the hair bundle are attenuated by adaptation of the mechanoelectrical transduction current and the voltage-dependent activation of a large potassium (K)-selective outwardly rectifying conductance, gDR. These processes may contribute to high-pass filtering of the responses of some utricular afferents to sinusoidal linear accelerations below 2 Hz. Depolarizing receptor potentials are more attenuated by gDR than are hyperpolarizing receptor potentials. It may therefore reduce nonlinear distortion introduced by mechanoelectrical transduction, which generates larger depolarizing currents than hyperpolarizing currents. The discharge properties of utricular afferents vary according to whether they innervate the striolar or extrastriolar zones of the sensory epithelium. Regional variation in hair-cell properties is likely to contribute. Preliminary results suggest that the outwardly rectifying K conductances of type II cells are slower and larger in the striola than in the extrastriola, consistent with regional variation in the relative numbers of delayed rectifier and A-current K channels.

Animals↗
PubMed1999

Sensory systems.

Explore the source record for details and available documents.

Animals↗
PubMed1998

Postnatal development of type I and type II hair cells in the mouse utricle: acquisition of voltage-gated conductances and differentiated morphology.

The type I and type II hair cells of mature amniote vestibular organs have been classified according to their afferent nerve terminals: calyx and bouton, respectively. Mature type I and type II cells also have different complements of voltage-gated channels. Type I cells alone express a delayed rectifier, gK,L, that is activated at resting potential. We report that in mouse utricles this electrophysiological differentiation occurs during the first postnatal week. Whole-cell currents were recorded from hair cells in denervated organotypic cultures and in acutely excised epithelia. From postnatal day 1 (P1) to P3, most hair cells expressed a delayed rectifier that activated positive to resting potential and a fast inward rectifier, gK1. Between P4 and P8, many cells acquired the type I-specific conductance gK,L and/or a slow inward rectifier, gh. By P8, the percentages of cells expressing gK,L and gh were at mature levels. To investigate whether the electrophysiological differentiation correlated with morphological changes, we fixed utricles at different times between P0 and P28. Ultrastructural criteria were developed to classify cells when calyces were not present, as in cultures and neonatal organs. The morphological and electrophysiological differentiation followed different time courses, converging by P28. At P0, when no hair cells expressed gK,L, 33% were classified as type I by ultrastructural criteria. By P28, approximately 60% of hair cells in acute preparations received calyx terminals and expressed gK,L. Data from the denervated cultures showed that neither electrophysiological nor morphological differentiation depended on ongoing innervation.

Age Factors↗
PubMed1998

Hair cells in mammalian utricles.

Two morphological classes of mechanosensory cells have been described in the vestibular organs of mammals, birds, and reptiles: type I and type II hair cells. Type II hair cells resemble hair cells in other organs in that they receive bouton terminals from primary afferent neurons. In contrast, type I hair cells are enveloped by large cuplike afferent terminals called calyces. Type I and II cells differ in other morphological respects: cell shape, hair bundle properties, and more subtle ultrastructural features. Understanding the functional significance of these strikingly different morphological features has proved to be a challenge. Experiments that correlated the response properties of primary vestibular afferents with the morphologies of their afferent terminals suggested that the synapse between the type I hair cell and calyx ending is lower gain than that between a type II hair cell and a bouton ending. Recently, patch-clamp experiments on isolated hair cells have revealed that type I hair cells from diverse species have a large potassium conductance that is activated at the resting potential. As a consequence, the voltage responses generated by the type I hair cells in response to injected currents are smaller than those generated by type II hair cells. This may contribute to the lower gain of type I inputs to primary afferent neurons. Studies of neonatal mouse utricles show that the type I-specific potassium conductance is not present at birth but emerges during the first postnatal week, a period of morphological differentiation of type I and type II hair cells.

Animals↗
PubMed1998

Ionic currents and electromotility in inner ear hair cells from humans.

The upright posture and rich vocalizations of primates place demands on their senses of balance and hearing that differ from those of other animals. There is a wealth of behavioral, psychophysical, and CNS measures characterizing these senses in primates, but no prior recordings from their inner ear sensory receptor cells. We harvested human hair cells from patients undergoing surgical removal of life-threatening brain stem tumors and measured their ionic currents and electromotile responses. The hair cells were either isolated or left in situ in their sensory epithelium and investigated using the tight-seal, whole cell technique. We recorded from both type I and type II vestibular hair cells under voltage clamp and found four voltage-dependent currents, each of which has been reported in hair cells of other animals. Cochlear outer hair cells demonstrated electromotility in response to voltage steps like that seen in rodent animal models. Our results reveal many qualitative similarities to hair cells obtained from other animals and justify continued investigations to explore quantitative differences that may be associated with normal or pathological human sensation.

Adult↗
PubMed1997

Mechanoelectrical transduction and adaptation in hair cells of the mouse utricle, a low-frequency vestibular organ.

Hair cells of inner ear organs sensitive to frequencies above 10 Hz adapt to maintained hair bundle deflections at rates that reduce their responses to lower frequencies. Mammalian vestibular organs detect head movements at frequencies well below 10 Hz. We asked whether hair cells of the mouse utricle adapt, and if so, whether the adaptation was similar to that in higher frequency organs such as the frog saccule. Whole-cell transduction currents were recorded from hair cells in the epithelium of the mouse utricle. Hair bundles were deflected by a fluid jet or a stiff probe. The transduction currents evoked by step deflections adapted over 10-100 msec. The mean operating range was 1.5 micron (deflection of the tip of the bundle), approximately threefold larger than in frog saccule. Taller and more compact bundles of the mouse utricle account for this difference. As in frog saccular hair cells, adaptation shifted the current-deflection (I(X)) relation along the deflection axis. These adaptive shifts had time constants of 10-20 msec and reached 60-80% of stimulus amplitude. The adaptive shift and voltage-dependent bundle movement are consistent with the motor model of adaptation. When the fluid jet was used, adaptation also broadened the I(X) relation and reduced the maximum current. Adaptation attenuated the transduction currents evoked by sinusoidal bundle deflections below 5 Hz, within the frequency range of the utricle, but because it was incomplete, substantial responses remained. Moreover, the adaptive shift mechanism preserves sensitivity even in the presence of large stimuli that would otherwise saturate transduction.

Adaptation, Physiological↗
PubMed1996

A delayed rectifier conductance in type I hair cells of the mouse utricle.

1. Membrane currents of hair cells in acutely excised or cultured mouse utricles were recorded with the whole cell voltage-clamp method at temperatures between 23 and 36 degrees C. 2. Type I and II hair cells both had delayed rectifier conductances that activated positive to -55 mV. 3. Type I, but not type II, hair cells had an additional delayed rectifier conductance (gK,L) with an activation range that was unusually negative and variable. At 23-25 degrees C, V(1/2) values ranged from -88 to -62 mV in 57 cells. 4. gK,L was very large. At 23-25 degrees C, the average maximum chord conductance was 75 +/- 65 nS (mean +/- SD, n = 57; measured at -54 mV), or approximately 21 nS/pF of cell capacitance. 5. gK,L was highly selective for K+ over Na+ (permeability ratio PNa+/PK+:0.006), but unlike other delayed rectifiers, gK,L was significantly permeable to Cs+ (PCs+/PK+:0.31). gK,L was independent of extracellular Ca2+. 6. At -64 mV, Ba2+ and 4-aminopyridine blocked gK,L with apparent dissociation constants of 2.0 mM and 43 microM, respectively. Extracellular Cs+ (5 mM) blocked gK,L by 50% at -124 mV. Apamin (100 nM) and dendrotoxin (10 nM) has no effect. 7. The kinetic data of gK,L are consistent with a sequential gating model with at least two closed states and one open state. The slow activation kinetics (principal time constants at 23-25 degrees C:600-200 ms) had a thermal Q10 of 2.1. Inactivation (Q10:2.7) was partial at all temperatures. Deactivation followed a double-exponential time course and had a Q10 of 2.0. 8. At 23-25 degrees C, gK,L was appreciably activated at the mean resting potential of type I hair cells (-77 +/- 3.1 mV, n = 62), so that input conductances were often more than an order of magnitude larger than those of type II cells. If these conditions hold in vivo, type I cells would produce unusually small receptor potentials. Warming the cells to 36 degrees C produced parallel shifts in gK,L's activation range (0.8 +/- 0.3 mV/degrees C, n = 8), and in the resting potential (0.6 +/- 0.3 mV/degrees C, n = 4). Thus the high input conductances were not an artifact of unphysiological temperatures but remained high near body temperature. It remains possible that in vivo gK,L's activation range is less negative and input conductances are lower; the large variance in the voltage range of activation suggests that it may be subject to modulation.

Animals↗
PubMed1995

Inwardly rectifying currents of saccular hair cells from the leopard frog.

1. Inwardly rectifying currents were characterized in sensory hair cells isolated from the saccules of leopard frogs, using the whole cell configuration of the patch-clamp technique in voltage-clamp mode. 2. Two types of inwardly rectifying currents were distinguishable based on their ionic selectivity, activation and deactivation kinetics, voltage dependence, dependence on external K+ and sensitivity to divalent cations. 3. One inwardly rectifying current displayed K+ selectivity, rapid monoexponential activation (tau approximately 1 ms at -120 mV), steep voltage dependence, dependence of the activation voltage range on external K+ and block by external Ba2+. We refer to this current as IK1, consistent with the terminology used for a similar current in cardiac cells. In 5 mM external K+, IK1 activated negative to -60 mV, was half-activated at -86 mV and fully activated by -110 mV. 4. The other inwardly rectifying current was a mixed K+/Na+ current with slow sigmoidal activation (slow tau approximately 100 ms at -120 mV) and deactivation, shallow voltage dependence and no dependence of the activation curve on external K+ and which was blocked by external Cd2+. This current was called Ih because of its similarities to Ih of photoreceptors. Ih activated negative to -50 mV, was half-activated at -90 mV and was fully activated at -130 mV. 5. A correlation between cell shape and the type of inwardly rectifying current was noted; the more spherical cells had Ih alone and the more cylindrically shaped cells had Ih and IK1. 6. The mean resting potential of 115 cells with IK1 and Ih was -68 +/- 0.5 mV (mean +/- SE) and that of 53 cells with Ih alone was -50 +/- 0.5 mV. This suggests that IK1 contributes to the more negative resting potential of the cylindrical cells. 7. In current-clamp mode, the voltage responses to current steps of the two cell populations differed. Small negative current steps evoked faster, smaller responses in cells with IK1 and Ih than in cells with Ih alone. In cells with Ih alone, long (> 100 ms) negative current steps evoked a hyperpolarization that partly repolarized as Ih activated. Cells with Ih alone showed electrical resonance at rest whereas cells with IK1 resonated only in response to positive current steps. 8. A model developed to explain electrical resonance in bullfrog saccular hair cells was adapted to include Ih or IK1 and Ih.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Potassium channels in squid neuron cell bodies: comparison to axonal channels.

The squid giant axon is formed from the fusion of many axons whose cell bodies are located in the giant fiber lobe (GFL) of the stellate ganglion. We measured macroscopic potassium channel currents in both squid giant axons and in the cell bodies. These currents appear similar in many ways, but were differently affected by the amino group modifying reagent, trinitrobenzene sulfonic acid (TNBS). TNBS increased the steady-state amplitude of the axonal currents but decreased the somal currents. We also studied single K channel currents in the giant fiber lobe cells, for comparison with single K channel currents measured previously in the axons. We found two common classes of K channels, distinguished by their conductances (11 and 25 pS) and kinetic properties. The two channel types resemble the 10 and 20 pS channels that have been described in squid giant axons. These results suggest that the cell bodies and axons express fundamentally similar K channel types. The different effects of amino group modification on macroscopic K currents in the two regions may arise from small (perhaps posttranslational) modifications of the core proteins.

Amines↗
PubMed1993

Electrical resonance of isolated hair cells does not account for acoustic tuning in the free-standing region of the alligator lizard's cochlea.

The cochlea of the alligator lizard is divided into two morphologically and physiologically distinct regions. In the "tectorial region," hair bundles of hair cells are draped by a tectorial membrane, whereas in the "free-standing region," hair bundles are said to be free-standing because there are no overlying tectorial structures. The acoustic tuning of the free-standing region depends at least in part on mechanical resonances of the hair bundles. In the turtle cochlea, in contrast, acoustic tuning depends in large part upon the electrical properties of the hair cells. We have investigated the electrical properties of hair cells isolated from the free-standing region of the alligator lizard's cochlea. When injected with steps of depolarizing current, these "free-standing hair cells" exhibited electrical resonances that were comparable in frequency and quality to electrical resonances in cochlear hair cells from turtles, chicks, and alligators, and in saccular hair cells from frogs and fish. In the lizard's free-standing hair cells, however, the electrical resonance frequencies (< 300 Hz) were a decade below the cells' acoustic characteristic frequencies (between 1 and 4 kHz), showing that the electrical resonance is not likely to contribute to acoustic tuning. The electrical resonances were not apparent at rest. The cells' resting potentials were significantly more negative than the activation voltage (approximately -40 mV) of the Ca(2+)-dependent K+ current upon which the electrical resonance has been shown to depend in other hair cells. At potentials more negative than -50 mV, an inwardly rectifying K+ conductance dominated. Because we observed no electrical tuning above 300 Hz, our results indirectly support a mechanical origin for acoustic tuning in the free-standing region of the alligator lizard cochlea. These results further show that acoustic tuning cannot be inferred solely from the electrical resonances of isolated hair cells.

Animals↗
PubMed1991

Induction of K-channel expression in a neuroblastoma cell line.

Whole-cell currents were examined in mouse neuroblastoma cells of the N2AB-1 line. In standard culture medium, N2AB-1 cells exhibited large voltage-dependent Na currents but no discernible K currents. Treatment of N2AB-1 cells with either dimethylsulfoxide (DMSO) in low-serum medium or with retinoic acid (RA) caused the expression of delayed rectifier K currents. Currents from two types of K channel with single channel slope conductances of 15.0 pS and 6.4 pS were observed in outside-out patches from cells of both treatment groups. Thus, while N2AB-1 cells did not exhibit K currents under standard culture conditions, they did possess the gene(s) encoding K channels. The treatments caused other changes that were not directly linked to K-channel expression. RA treatment caused neurite extension in most, but not all, N2AB-1 cells; however, all RA-treated cells, including those without neurites, expressed K currents. RA treatment did not suppress cell division or cause hypertrophy. In contrast, treatment with DMSO/low serum suppressed cell division and caused cellular hypertrophy, but did not cause long neurites to form. Thus, the regulation of K channels was not coupled in a simple fashion to properties that have been associated with a differentiated neuronal phenotype: neurite elaboration, changes in cell size, and inhibition of cell division. These results suggest that N2AB-1 cells may be a good model system for investigating the processes regulating K-channel expression.

4-Aminopyridine↗