Multiple modes of inner hair cell stimulation.

Most current theories of cochlear mechanics assume that the pattern of cochlear partition vibration is simple, similar to that of a bending beam. Recent evidence suggests, however, that the vibration of the organ of Corti can be complex and that multiple vibrational modes may play an important role in cochlear transduction. Inner hair cell (IHC) and auditory nerve responses to pure tones can exhibit large phase shifts and complex response waveforms with increasing stimulus level. In contrast, the comparable basilar membrane (BM) responses are much less complex, exhibiting only small phase shifts and relatively sinusoidal waveforms. To reconcile the differences observed between the published BM data and the IHC data, we have recorded receptor potentials from IHCs and compared these waveform data to the output of two computational models: a traditional linear model where IHC excitation depends only on BM displacement and a new model that assumes that outer hair cell (OHC) force production provides the major mechanical input to the IHC along with two additional mechanical components. Comparisons of the output of the two models with the experimental data show that the new model is capable of reproducing the very complex voltage responses of the IHC recorded in vivo whereas the traditional model performed poorly.

Acoustically activated c-fos expression in auditory nuclei of the anaesthetised guinea pig.

The spatial expression of the immediate-early gene c-fos in central auditory nuclei of the anaesthetised guinea pig was investigated following exposure of the animal to acoustic stimulation. Accurate control of both the spectra and the level of the stimulus was designed so that the presumed excitation of central auditory nuclei was similar across animals. For unstimulated anaesthetised control animals levels of labelling were significantly higher when compared with unanaesthetised controls. This appeared to a result of the combination of the experimental manipulations and also the use of the anaesthetic. A surprising finding was that unstimulated control animals placed in an anechoic chamber demonstrated the highest levels of fos-like immunoreactivity (Fos-LIR). When anaesthetised animals were exposed to acoustic stimuli the total number of cells showing Fos-LIR was elevated when compared to anaesthetised, but unstimulated animals. There was no evidence at any level of the auditory pathway that these animals demonstrated spatially restricted Fos-LIR which may have suggested place-frequency mapping. In contrast, spatially restricted labelling was found in awake animals exposed to an identical stimulus.

Transfer characteristic of the inner hair cell synapse: steady-state analysis.

Inner hair cells (IHC) transduce mechanical to electrical energy in the mammalian cochlea producing a receptor potential which is a rectified, filtered representation of the mechanical input to the hair cell. The IHC synapse transfers the information in the receptor potential to the fibers of the auditory nerve (whose cell bodies form the spiral ganglion) where it is encoded as a pattern of action potentials. That transfer was investigated by comparing the steady-state responses in pre- and post-synaptic cells. A nonlinear transfer characteristic describing the synapse was generated by plotting the spiral ganglion cell firing rate as a function of the IHC receptor potential. For each spiral ganglion unit, the operating range maps onto a different portion of the nonlinear inner hair cell operating range, dependent on the neural unit's threshold. Units whose rate-level functions exhibit similar slopes but different thresholds can have dramatically differing sensitivities to changes in the IHC potential. This threshold-dependent mapping supports the concept that information may be distributed amongst nerve fibers according to their threshold.

Acoustic lesions in the mammalian cochlea: implications for the spatial distribution of the 'active process'.

The spatial contribution of mechanically active hair cells to tuning and sensitivity at a single point in the mammalian cochlea has been investigated in the basal turn of the guinea pig cochlea. Following the destruction of outer hair cells with acoustic overstimulation it was possible to record apparently normal tuning and sensitivity from spiral ganglion neurones innervating inner hair cells located on the apical edges of substantial lesions. The distance between the recording site, where neurones showed normal sensitivity, and areas of the cochlea showing 60-100% of the outer hair cells either damaged or missing varied between 0.2 and 1.3 mm which incorporates approximately 70 to 450 outer hair cells. In one animal neurones that demonstrated normal sensitivity were recorded within 0.2 mm of a lesion where 67% of the outer hair cells were either missing or showed severe damage to their stereocilia and within 0.5 mm of areas of the organ of Corti showing damage to 97% of the outer hair cells. This distance includes approximately 50 inner hair cells or 180 outer hair cells. The location of these neurones, whose sharp tuning presumably mirrors basilar membrane mechanics, suggests that a substantial proportion of point tuning in the cochlea may be derived over a distance of less than 0.5 mm and involve fewer than 200 active outer hair cells.

Low-frequency responses of inner hair cells: evidence for a mechanical origin of peak splitting.

Auditory nerve fibres usually respond to a preferred phase of a low frequency sinusoidal stimulus. However, at high sound pressures fibres may either change the preferred phase of response or respond to more than one phase of the stimulus. This complex firing pattern is known as peak splitting. Hypotheses for the origin of peak splitting have ranged from micromechanical models to models incorporating electrical interactions between inner and outer hair cells. In order to determine the origin of peak splitting, the potential across the IHC synaptic membrane has been measured during stimulation with low frequency tones and it is found that the IHC receptor potentials exhibit peak splitting at sound pressures that coincide with the saturation of the outer hair cell receptor potentials. Current injection experiments show that peak splitting is also recorded in the resistance change of the inner hair cell during acoustic stimulation. It is concluded from this evidence that peak splitting is present in the mechanical input to the IHC.

Acoustically induced hearing loss: intracellular studies in the guinea pig cochlea.

Intracellular recordings were made from both inner and outer hair cells (IHC, OHC) in the basal coil of the guinea pig cochlea before, during and after the animal was exposed to loud, pure tones. Following multiple loud tones both types of sensory cells demonstrate a culmulative decrease in their voltage responses to a test tone. A loss in sensitivity of the compound action potential (CAP) of the eighth nerve co-incides with a decrease in both the amplitude of the IHC receptor potential and the positive summating potential (+SP) recorded at the round window. The largest decreases in sensitivity of IHCs are found at the characteristic or best frequency (CF) of each cell and this frequency selective loss of sensitivity results in a decrease in the tuning of the hair cell. During and following a loud tone the nonlinear properties of IHCs are also reduced.

The response of hair cells in the basal turn of the guinea-pig cochlea to tones.

1. Intracellular recordings were made from inner and outer hair cells in the basal turn of the guinea-pig cochlea. The resting membrane potentials of the inner hair cells are more positive than -50 mV while those of outer hair cells are usually more negative than -70 mV. 2. At low frequencies the receptor potentials of inner hair cells are predominantly depolarizing while those from outer hair cells are hyperpolarizing at low and moderate sound pressure (e.g. less than 90 dB re 2 X 10(-5) Pa at 600 Hz). The potentials then become predominantly depolarizing at high sound pressure. 3. The asymmetry of the inner and outer hair cell receptor potentials are manifested instantaneously except at high stimulus levels when the depolarizing responses of outer hair cells take several cycles to develop. 4. At the offset of intense tones outer hair cell membrane potentials remain depolarized by 1-2 mV above their resting value and return to normal over a period depending on the level and duration of the tone. 5. In response to tones above about 2 kHz and at levels below about 90 dB the wave forms of outer hair cell receptor potentials are virtually symmetrical without measurable d.c. components. In response to tones close to their best frequencies (16-21 kHz), inner hair cells in the basal turn generate large depolarizing (d.c.) receptor potentials while outer hair cells from this region of the cochlea do not generate significant voltage responses. 6. Frequency tuning curves were derived for inner and outer hair cells from the amplitude-intensity relationships of their d.c. and phasic (a.c.) receptor potentials respectively. When the latter were compensated for the low-pass characteristics of the recording system and the hair cell time constant, the frequency selectivity of inner and outer hair cells are similar. 7. The response properties of inner and outer hair cells in the basal turn of the guinea-pig cochlea are discussed in relation to their proposed roles in mechano-electric transduction.

Mechanosensitivity of mammalian auditory hair cells in vitro.

Intracellular responses recorded in vitro from the cochleas of anaesthetized mammals have shown that the mechanoreceptive inner and outer hair cells are sharply tuned, accounting for many of the properties of the afferent fibres in the auditory nerve. However, in vivo it has not been possible to measure directly the excitatory mechanical input to these cells (the displacement of their mechanosensitive stereocilia) and thus to determine the relationship between the receptor potentials and displacement of their stereocilia. As a means of circumventing this technical difficulty, we have developed an organ culture of the mouse cochlea and here we describe the receptor potentials generated by the hair cells in response to direct displacement of their stereocilia.

The responses of inner and outer hair cells in the basal turn of the guinea-pig cochlea and in the mouse cochlea grown in vitro.

Until recently the responses of the mechanosensitive hair cells of the cochlea have been inferred from their morphology, morphological relationships with other structures in the cochlea, and by indirect electrophysiological measurements. With the advent of techniques for making intracellular recordings from hair cells in the cochleas of anaesthetised mammals it has been possible to measure the responses of hair cells to acoustic stimulation and to assess their roles in sensory transduction in the cochlea. Intracellular recordings of the responses of inner and outer hair cells in the basal turn of the guinea-pig cochlea show that they differ considerably from each other. The receptor potentials of inner hair cells are larger, predominantly depolarizing to low frequency tones and at their best frequencies (16-20 kHz) they generate depolarizing dc receptor potentials. Outer hair cells generate predominantly hyperpolarizing potentials to low frequency tones. They do not produce significant voltage responses at high frequencies except at high intensities when they generate slowly rising depolarizing potentials which are associated with loss of cochlear sensitivity. At low frequencies the receptor potentials of the inner hair cells phase lead those of the outer hair cell. Measurements of their frequency selectivity show that inner and outer hair cells are both sharply tuned. It is proposed that the responses of inner and outer hair cells are consistent with sensory and motor roles respectively in mechanoelectric transduction and that the outer hair cells are the site of an active mechanical process responsible for the frequency selectivity and sensitivity of the cochlea. Intracellular recordings from hair cells in the mouse cochlea maintained in vivo have provided a direct measure of the mechanosensitivity of cochlear hair cells (approximately 30 mV per degree of displacement of their stereociliary bundles) and indirect evidence that the transfer characteristics of the outer hair cells in vivo may be due to their mechanoelectrical interaction with the tectorial membrane. This is because the transfer characteristics of the inner and outer hair cells are similar in vitro in the absence of a tectorial membrane. Considerable importance is attributed to the shape of the transfer characteristics of the inner and outer hair cells. Changes in these characteristics during anoxia and following exposure to intense tones are associated with depolarization of the outer hair cells and loss of cochlear sensitivity and frequency selectivity. Current-voltage studies of hair cells in vivo show the inner and outer hair cells to be electrically nonlinear.(ABSTRACT TRUNCATED AT 400 WORDS)

Outer hair cells in the mammalian cochlea and noise-induced hearing loss.

Hair cells in the mammalian cochlea transduce mechanical stimuli into electrical signals leading to excitation of auditory nerve fibres. Because of their important role in hearing, these cells are a possible site for the loss of cochlear sensitivity that follows acoustic overstimulation. We have recorded from inner and outer hair cells (IHC, OHC) in the guinea pig cochlea during and after exposure to intense tones. Our results show functional changes in the hair cells that may explain the origin of noise-induced hearing loss. Both populations of hair cells show a reduction in amplitude and an increase in the symmetry of their acoustically evoked receptor potentials. In addition, the OHCs also suffer a sustained depolarization of the membrane potential. Significantly, the membrane and receptor potentials of the OHCs recover in parallel with cochlear sensitivity as measured by the IHC receptor potential amplitude and the auditory nerve threshold. Current theories of acoustic transduction suggest that the mechanical input to IHCs may be regulated by the OHCs. Consequently, the modified function of OHCs after acoustic overstimulation may determine the extent of the hearing loss following loud sound.

Recovery of eighth nerve action potential thresholds after exposure to short, intense pure tones: similarities with temporary threshold shift.

Exposure of guinea pig cochleas to short (25, 250 and 1000 ms), intense (90 or 100 dB SPL) 10 kHz pure tones reduced cochlear sensitivity to 14 kHz test tones presented at intervals varying from 5 ms to 70 s after the exposure tone. The recovery of cochlear sensitivity, determined as the SPL required to evoke a 20 microV compound action potential (CAP), depended on both the intensity and the duration of the exposure tone and appeared to take place in two or more phases. After a 25 ms, 100 dB SPL exposure, CAP threshold increased by up to 13.5 dB and generally recovered very rapidly (25 ms), although some loss persisted for as long as 400 ms. A similar, but greater and longer, elevation of threshold was seen after long exposure tones. Lower exposure tone intensities (90 dB SPL) produced threshold elevations which generally lasted for only short durations (25-35 ms). The rapid recovery is consistent with the time course of recovery from rapid adaptation, while the slow recovery component is similar to that seen in the cochlea after much longer exposures and may be related to the phenomenon of temporary threshold shift. Long and/or loud exposures also frequently resulted in a third form of threshold elevation, identified in only a few animals, which recovered with a time constant in excess of 25 s. A fourth component occasionally persisted for the duration of the experiment.

Variability of noise-induced damage in the guinea pig cochlea: electrophysiological and morphological correlates after strictly controlled exposures.

Anaesthetized guinea pigs were exposed to loud tones (1 h, 10 kHz, 112-118 dB SPL) with continuous control of the sound pressure at the tympanic membrane. N1 electrocochleograms were used to measure functional damage immediately and 21 days after the exposure. Damage to the organ of Corti was assessed by scanning electron microscopy and light and transmission electron microscopy. Principal findings were: (1) Functional impairment after 21 days showed large inter-animal variation which was not the result of changes in the effective damaging energy. (2) Structural damage to the stereocilia was also variable and did not always correlate with functional impairment, although when N1 thresholds were elevated damage to the stereocilia was always present. (3) Unknown factors within the cochlea must be responsible for variations in individual susceptibility to permanent noise-induced hearing loss.

Reduced temporary and permanent hearing losses with multiple tone exposures.

Acoustic overstimulation of the guinea pig cochlea with a 16 kHz pure tone induces a loss in threshold sensitivity that can be either temporary or permanent depending on the duration of the trauma. When a second tone of lower frequency (10, 5 or 2 kHz) is presented to the same cochlea simultaneously with the first tone, then the resultant threshold loss produced by the 16 kHz tone is significantly less. This applies to both temporary and permanent threshold losses. The reduced threshold loss disappears as the intensity of the second tone decreases. This type of nonlinear cochlea behaviour is similar to other acoustically evoked nonlinearities generally grouped under the term, two-tone suppression or inhibition. The results disagree with the "equal energy hypothesis' as a method to establish damage risk criteria in noise-induced hearing loss.

Temporary threshold shift modified by binaural acoustic stimulation.

Monaural losses in hearing sensitivity induced by an intense pure tone could be reduced if an acoustic stimulus of the same frequency was simultaneously delivered to the other ear. The reduction was eliminated when the contralateral stimulus was set at a frequency other than the ipsilateral trauma frequency and also after the administration of strychnine, a known blocker of auditory efferent activity. This suggests that acoustic activation of auditory efferents is responsible for the reduced ipsilateral sensitivity loss.

Acoustic trauma: single neuron basis for the "half-octave shift".

Exposure to an intense pure tone can induce a loss of hearing sensitivity. If this loss recovers, then the desensitization is regarded as a temporary threshold shift (TTS). At the single auditory neuron level this TTS was monitored as a loss of sensitivity at the neuron's most sensitive or characteristic frequency (CF). When pure-tone exposures were presented at frequency intervals measured from the neuron CF, then a frequency half an octave below the CF was the most effective for inducing a CF TTS. All exposure frequencies higher than half an octave below the CF produce a marked reduction in TTS growth with intensity, when compared to lower exposure frequencies. This behavior is such that, with increasing exposure frequency higher than the -1/2-octave point, the intensity needed to produce a given TTS grew faster than the neuron sensitivity. However, below the -1/2-octave point all exposure frequencies were similarly behaved. Strong similarities exist between the frequency-specific requirements for TTS and the mechanical and neural nonlinearities found in other studies. This suggests that the half-octave shift may well be a direct result of basilar membrane nonlinearities.

Single auditory neuron response during acute acoustic trauma.

By recording from single auditory neurons in the spiral ganglion of the guinea pig cochlea it was possible to monitor threshold changes to acoustic stimuli in the same neuron during acute exposures to continuous pure tones (100 dB SPL) delivered for periods of up to 180 min. Initial changes in the tuning curve were sensitivity losses at and above the characteristic frequency (CF). This asymmetric loss resulted in a shift in the CF to lower frequencies. Further exposure (up to 20 min) produced a complete loss of the sharply tuned portion of the tuning curve and the most sensitive area moved to lower frequencies by up to 0.75 of an actave when compared to the original CF. Sensitization of the low frequency section of the tuning curve could take place for short exposures (up to 20 min) but disappeared with further exposure, whether continuous or interrupted. Long term exposures (60--80 min) produced losses in the low frequency portion of the tuning curve until a point was reached when the unit no longer responded to stimuli at 110 dB SPL.

Electrophysiological and morphological changes in the guinea pig cochlea following mechanical trauma to the organ of Corti.

Small discrete lesions were produced in the organ of Corti of the guinea pig cochlea using fine probes to produce direct mechanical insult. The electrophysiological state of the cochlea was assessed using N1 electrocochleography and loss of receptor cells determined by scanning electron microscopy. Principal findings were: 1) Excellent agreement between the location of hair cell losses and the frequency of maximum sensitivity change in the N1 audiogram; 2) The spatial extent of the mechanically induced lesion appears to be more important than the total number of hair cells lost, in determining the magnitude of N1 sensitivity loss; 3) Hair cell losses extending over only 72 micrometers could be detected as significant changes in N1 sensitivity. These results further emphasize the accuracy and usefulness of the N1 electrocochleogram for assessing the functional status of the cochlea; 4) Lesions involving only outer hair cell loss also produced marked elevations of N1 threshold.