Degeneration in the cochlea after noise damage: primary versus secondary events

PURPOSE: To determine if noise damage in the organ of Corti is different in the low- and high-frequency regions of the cochlea. MATERIALS AND METHODS: Chinchillas were exposed for 2 to 432 days to a 0.5 (low-frequency) or 4 kHz (high-frequency) octave band of noise at 47 to 95 dB sound pressure level. Auditory thresholds were determined before, during, and after the noise exposure. The cochleas were examined microscopically as plastic-embedded flat preparations. Missing cells were counted, and the sequence of degeneration was determined as a function of recovery time (0-30 days). RESULTS: With high-frequency noise, primary damage began as small focal losses of outer hair cells in the 4-8 kHz region. With continued exposure, damage progressed to involve loss of an entire segment of the organ of Corti, along with adjacent myelinated nerve fibers. Much of the latter loss is secondary to the intermixing of cochlear fluids through the damaged reticular lamina. With low-frequency noise, primary damage appeared as outer hair cell loss scattered over a broad area in the apex. With continued exposure, additional apical outer hair cells degenerated, while supporting cells, inner hair cells, and nerve fibers remained intact. Continued exposure to low-frequency noise also resulted in focal lesions in the basal cochlea that were indistinguishable from those resulting from exposure to high-frequency noise. CONCLUSIONS: The patterns of cochlear damage and their relation to functional measures of hearing in noise-exposed chinchillas are similar to those seen in noise-exposed humans. Thus, the chinchilla is an excellent model for studying noise effects, with the long-term goal of identifying ways to limit noise-induced hearing loss in humans.     

Cochlear hair cell stereocilia loss in LP/J mice with bone dysplasia of the middle ear

LP/J inbred mice spontaneously develop bony lesions of the middle ear and otic capsule that are similar to those of human otosclerosis and tympanosclerosis. These mice also have progressive loss of hearing due to cochlear hair cell loss. The purpose of this study was to describe quantitatively the deterioration and loss of cochlear hair cells to serve as a basis for future experiments attempting to alter the course of this disorder. Cochleas from 37 LP/J inbred mice were examined by scanning electron microscopy. The stereocilia loss in the cochlea was evident as early as 15 weeks of age and progressed from the basal turn to the apex. Outer hair cells were affected more than inner hair cells. As outer hair cells deteriorated we observed fusion, bending, and breakage of stereocilia. There were no apparent differences in the mode of deterioration among the three rows of outer hair cells. Stereocilia fusion of inner hair cells occurred at an older age, and giant, elongated stereocilia were found in some of the animals.     

Differential vulnerability of basal and apical hair cells is based on intrinsic susceptibility to free radicals

The base of the cochlea is more vulnerable to trauma than the apex as seen in the pattern of hair cell damage by cisplatin or aminoglycosides. The differential vulnerability is maintained in organotypic cultures exposed directly to these drugs, suggesting there may be an intrinsic difference in sensitivity to damage along the cochlear spiral. We therefore investigated the survival capacity of isolated outer hair cells and strips dissected from different turns of the guinea pig organ of Corti in short-term culture. Cells were stained with fluorescent indicators of viable or dead cells, calcein-AM and ethidium homodimer. After 5 h at room temperature, up to 90% of outer hair cells from the apex survived, but less than 30% from the base. In contrast, basal inner hair cells remained viable, and supporting cells survived for at least 20 h. The difference in survival capacity between basal and apical outer hair cells coincided with a significantly lower level of the antioxidant glutathione in basal outer hair cells compared with apical outer hair cells. This suggested that basal outer hair cells may be more vulnerable to free-radical damage than apical outer hair cells. The survival of basal outer hair cells was significantly improved by addition of the radical scavengers n-acetyl cysteine, p-phenylenediamine, glutathione, mannitol or salicylate. The protection by antioxidants implies that the accelerated death of basal outer hair cells is due to free-radical damage. The results support an intrinsic susceptibility to free radicals that differs among cochlear cell populations. This differential provides a rational explanation for base-to-apex gradients observed in various forms of cochlear pathology.     

Psychophysical Evaluation of Cochlear Hair Cell Damage Due to the A3243G Mitochondrial DNA Mutation

Mitochondrial dysfunction is an important cause of human deafness, implicated in genetic deafness, toxin and noise damage. We assessed the mechanism of cochlear dysfunction in a population of 11 subjects with a specific mitochondrial disorder caused by the A3243G mitochondrial DNA mutation. Psychophysical tests were carried out to assess the inner and outer hair cell functions in vivo. Inner hair cell function was assessed using a measure of hearing threshold in the presence of "threshold-equalizing noise" which can indicate "dead regions" where the transduction mechanism fails. Outer hair cell function was assessed by using the notched-noise method to measure auditory filter width, dependent on active mechanisms in the outer hair cell. The results support the conclusion that this mitochondrial disorder causes both inner and outer hair cell dysfunctions. Evidence of inner hair cell dysfunction was found mainly in basal (high frequency) regions of the cochlea and occurred even in some subjects with only mild hearing loss. Evidence of outer hair cell dysfunction was found in some instances where pure tone threshold was at or close to normal. The common occurrence of dead regions in the basal cochlea has treatment implication for this form of deafness; such people may not be helped by amplification of high frequencies.     

Speech perception and otoacoustic emissions by pre-processing sound in the inner ear]

Direct observations of the basilar membrane movements show that sound perception can no longer be regarded as a passive process: vulnerable, energy-consuming amplification processes are required in the cochlea. The outer hair cells (OHC) fulfil this demand morphologically and functionally. These sensory cells have a double role: they perceive sound and thus modulate the cochlear biomechanics through their motile activity. The key event of sound transduction is performed by the inner hair cells (IHC) after active sound amplification in the OHC. The control of the OHC is assured by the efferent olivocochlear fibres which release acetylcholine (ACh) and gamma-aminobutyric acid (GABA) into the synaptic cleft at the basal pole of the OHC. Nicotinergic acetylcholine and GABA receptors within the outer cell membrane of OHC were identified and characterised. The application of the neurotransmitter GABA to the basal pole of vital OHC leads to a reversible elongation of the cylindrical cell body while ACh induces a reversible, slow contraction of the sensory cells. These two neurotransmitters are supposed to counteract in the control of the cochlear amplifier. The reciprocal distribution of ACh and GABA receptors and their counteracting function (contraction vs elongation) has an additional impact on the modulation of OHC function. The result is an even more diversified control of the cochlear amplifier. The energy-consuming cochlear amplifications are reflected by an epiphenomenon, i.e. the otoacoustic emissions (OAE). These are emitted by the cochlea and can be divided into "spontaneous OAE", "transitory evoked OAE" (TEOAE), "stimulus frequency OAE" and "distortion product OAE". The TEOAE are now an integrated part of audiological diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relation between changes in compound action potential tuning curves and the pathology of cochlear hair cells stereocilia]

Relations of the changes of compound action potential tuning curve (AP-TC) and AP(N1) threshold to the pathology of the cochlear hair cells stereocilia were examined in acoustic-traumatized guinea pigs. The results showed that all animals with hair cell stereocilia damaged showed deteriorations of AP-TC. It supports the suggestion that mechanical properties of hair cell stereocilia contribute to the frequency selectivity of cochlea. After exposure, the characteristic frequency (CF) shift of AP-TC was observed. The mechanism of CF shift may be related to the lesion of hair cell stereocilia which may change normal tono-topic on cochlear level. Correlation between the damages of inner/outer hair cell stereocilia and changes of AP-TC and AP threshold suggested that the threshold and tuning properties of the cochlea are dependent upon the integrity of the inner and outer hair cells and their inter-relationship.     

Chinchilla models of selective cochlear hair cell loss

Although it is well known that ethacrynic acid (EA) can enhance gentamicin (GM) ototoxicity, there has been no systematic study of the relationship between dosing parameters and inner ear pathology. We examined the effects of two parameters, GM dose and time delay between GM and EA administration, on cochlear and vestibular hair cell loss in chinchillas. 'No delay' groups received one injection of GM (125, 40, 20, or 10 mg/kg i.m.) followed immediately by EA (40 mg/kg i.v.); 'delay' groups received GM (10 mg/kg i.m.) followed by EA 1 or 1.5 h later. Animals were sacrificed 7 days later for evaluation of hair cell loss in the cochlea and vestibular end organs (cristae, saccule and utricle). Vestibular function was assessed prior to sacrifice by measuring the duration of nystagmus induced by cold caloric stimulation. No delay groups had approximately 100% loss of outer hair cells and dose-dependent losses of inner hair cells, ranging from approximately 100% to 58%. In 1 and 1.5 h delay groups, inner hair cell losses were approximately 19% and 0%, outer hair cell losses were approximately 74% and 47%, and outer hair cell loss followed a typical base to apex gradient. Two results were remarkable. First, the three groups with partial inner hair cell loss showed an atypical lesion pattern in which losses were substantially greater in the apical half than in the basal half of the cochlea. Second, there was no vestibular pathology in any group. The results establish dosing parameters that can be used to produce animal models with defined patterns and magnitudes of cochlear hair cell damage, but normal vestibular function and morphology.     

Structural effects of short term and chronic extracochlear electrical stimulation on the guinea pig spiral organ

To assess the effects of extracochlear electrical stimulation on cochlear structure, guinea pigs were implanted and stimulated with single middle ear electrodes either at round window or promontory sites, and their cochleae examined by transmission electron microscopy. Implanted but unstimulated, or unimplanted control animals were examined in the same way. Alternating current stimulation at the promontory for 2 h at 150 Hz, 500 microA, caused outer hair cell efferent endings to become dense and vacuolated, but no hair cells were damaged. With direct current stimulation at 500 microA for 2 h the basal regions of the stimulated cochlea were badly damaged and many outer hair cells lysed. Long term (up to 1200 h) round window stimulation at 100 or 141 Hz, 15-91 microA rms, did not cause cell death or inner hair cell damage, but basal outer hair cells and their efferent endings were badly affected in both ipsilateral and contralateral cochleae. The compound action potential of the auditory evoked response to broad band click stimuli was not altered by chronic electrical stimulation. It is concluded that chronic stimulation with the parameters used does not threaten cochlear survival, and it is proposed that the bilateral structural changes induced by chronic stimulation are caused by excessive activation of the cochlear efferent pathways.     

Selective destruction of the outer hair cells in the chinchilla

In the study of cochlear function an animal model in which apical outer hair cells could be destroyed with retention of inner hair cells would be valuable. Our object was to attempt to reproduce a reported method in a scientific manner. Behavioural response thresholds to pure tones were recorded in 7 monaural chinchillas. In 5, Kanamycin was administered in ototoxic dosage until the thresholds deteriorated. The other 2 served as controls. Cochlear histology was assessed from embedded surface preparations. In only one of the 5 animals was the aim achieved. In this animal outer hair cells were absent virtually throughout the cochlea and inner hair cell retention was basal. The thresholds were elevated on average 40 dB throughout the frequency range. This is in keeping with current concepts of cochlear function.     

Dimensions of the cochlear stereocilia in man and the guinea pig

The tuning properties of the basilar membrane and the presence of acoustic emissions from the cochlea suggest that an energy consuming, mechanically active cochlear amplifier exists. Some models of this amplifier demand a mechanical resonator within the cochlea. The lengths of the stereocilia of the inner and outer hair cells in man and the guinea pig have been measured from scanning electron micrographs using a stereometric technique. In both species there is a linear increase in the length of the longest inner hair cell stereocilia with distance along the cochlea. There are, however, marked differences between the dimensions of the outer hair cell stereocilia in the two species. In man, there is an increase in length which is is a hyperbolic function of distance along the cochlear duct. The picture is more complicated in the guinea pig. This could account for some of the differences in auditory physiology between the two groups. The mechanical resonance properties of the human OHC stereocilia have been assessed, and, with certain assumptions, these properties are such that resonance of the stereocilia of the OHCs could form part of the cochlea amplifier, at least in man.     

Changes in cochlear microphonic response after Y-ray irradiation of the inner ear of the guinea-pig

The effects of ionizing irradiation on the cochlear microphonic response of guinea-pigs were studied. The cochlear microphonics (CM) of both ears were recorded in a total of 36 animals. Recording was carried out by the differential electrode technique on the basal turn of the cochlea. One week prior to recording, the left ear of each animal had been exposed to 35-70 Gy radiation in increments of 5 Gy. Doses of 40 Gy and above led to a reduction in CM response. After doses of 60 Gy or more, no CM response could be detected. Damage most probably occurred in the outer and inner hair cells.     

Mechanical properties of sensory and supporting cells in the organ of Corti of the guinea pig cochlea--study by atomic force microscopy

Mammalian hearing is refined by amplification of the motion of the cochlear partition. To understand the cochlear amplification, mechanical models of the cochlea have been used. When the dynamic behavior of the cochlea is analyzed by a model, elastic properties of the cells in the organ of Corti must be determined in advance. Recently, elastic properties of outer hair cells (OHCs) and pillar cells have been elucidated. However, those of other cells have not yet been clarified. Therefore, in this study, using an atomic force microscope (AFM), elastic properties of Hensen's cells, Deiters' cells and inner hair cells (IHCs) in the apical turn and those in the basal and second turns were estimated. As a result, slopes indicative of cell elastic properties were (8.9 +/- 5.8) x 10(3) m(-1) for Hensen's cells (n = 30), (5.5 +/- 5.3) x 10(3) m(-1) for Deiters' cells (n = 20) and (3.8 +/- 2.6) x 10(3) m(-1) for IHCs (n = 20), and Young's modulus were 0.69 +/- 0.45 kPa for Hensen's cells and 0.29 +/- 0.20 kPa for IHCs. There was no significant difference between elastic properties of each type of cell in the apical turn and those in the basal and second turns. However, it was found that there is a significant difference between Young's moduli of cells estimated in this study and those of the OHCs and pillar cells reported previously.     

Cisplatin-induced increases in spontaneous neural activity in the dorsal cochlear nucleus and associated outer hair cell loss

Tinnitus is one of the consequences of cisplatin chemotherapy, but its underlying mechanisms are not well understood. Since it has been shown that cisplatin causes outer hair cell loss, it is possible that loss of these cells might induce tinnitus by increasing spontaneous activity in the central auditory system. To test this possibility, the present study examined the effects of cisplatin treatment on cochlear hair cells and on spontaneous neural activity in the dorsal cochlear nucleus of hamsters. Recordings, carried out approximately 1 month after cisplatin treatment, demonstrated significant increases in spontaneous activity across broad regions of the dorsal cochlear nucleus relative to levels in saline-treated controls. Histological results showed that cisplatin-treated animals also displayed dramatic loss of outer hair cells over most of the basal turn of the cochlea. Inner hair cells remained intact, although some evidence of damage to their stereocilia was evident. These findings indicate that cisplatin treatment causes abnormalities in spontaneous activity in the dorsal cochlear nucleus that are associated with widespread damage to outer hair cells. However, since some damage to inner hair cells was also observed, the role of inner hair cell injury in contributing to higher spontaneous activity cannot be ruled out.     

Cochlear hair cell loss in ears with cholesteatomas. Scanning electron microscopy study

Cochleas from 16 Mongolian gerbils with spontaneous aural cholesteatomas, and four of similar age without cholesteatomas, were examined by scanning electron microscopy to quantify cochlear hair cell loss. Loss of hair cell stereocilia was found in all ears with cholesteatomas and was significantly increased when compared with uninvolved ears from animals of similar age. The hair cell loss associated with gerbilline cholesteatomas appeared to be most marked in the middle turn of the cochlea and increased in severity with increasing size of the cholesteatomas. Outer hair cells were affected more than inner hair cells. Inner and outer hair cell loss was not significantly different in infected cholesteatomas versus sterile cholesteatomas. The greater damage to hair cells at the middle turn compared to the basal turn suggests that these losses may be the result of some agent acting through the cochlear wall rather than through the round window.     

Tonotopic Morphometry of the Lamina Reticularis of the Guinea Pig Cochlea with Associated Microstructures and Related Mechanical Implications

Morphometry of the lamina reticularis of the guinea pig cochlea was performed using scanning electron microscopy. Seventy-four geometrical parameters of the lamina reticularis, the bundles of stereocilia, and individual stereocilia, in all rows of hair cells and within the individual hair cells, were measured at ten equally spaced locations along the longitudinal direction of the cochlea. Variations of the parameters versus the longitudinal coordinate were statistically analyzed and fitted with polynomials (constant, linear, or quadratic). Our data show that a unique set of geometrical parameters of inner and outer hair cells is typical for every frequency-dependent position at the lamina reticularis. Morphology of the outer hair cell structures varies more than respective parameters of the inner hair cells. Mechanical modeling using the obtained geometrical parameters provides a novel glance at the mechanical characteristics with respect to the cochlear tonotopy.     

Glutamate is the Afferent Neurotransmitter in the Human Cochlea

Glutamate, the most important afferent neurotransmitter in the auditory system, is thought to be the afferent transmitter between the cochlear inner hair cells and afferent neurons, hitherto visualized only in the cochlea of animal species. It has been identified for the first time in sections from the human inner ear. L-glutamate, NMDAR2B and the enzyme glutamine synthetase were identified by using monoclonal antibodies. The distribution pattern of the transmitter L-glutamate in the human cochlea is similar to that observed in other mammals. L-glutamate was identified adjacent to outer and inner hair cells and in the spiral ganglion. Similar distributions were found for glutamine synthetase and the ionotropic NMDA receptor subunit NMDAR2. The identification of neurotransmitters and their receptors in the human cochlea has implications for the pharmacotherapy of inner ear diseases.     

Glutamate is the afferent neurotransmitter in the human cochlea

Glutamate, the most important afferent neurotransmitter in the auditory system, is thought to be the afferent transmitter between the cochlear inner hair cells and afferent neurons, hitherto visualized only in the cochlea of animal species. It has been identified for the first time in sections from the human inner ear. L-glutamate, NMDAR2B and the enzyme glutamine synthetase were identified by using monoclonal antibodies. The distribution pattern of the transmitter L-glutamate in the human cochlea is similar to that observed in other mammals. L-glutamate was identified adjacent to outer and inner hair cells and in the spiral ganglion. Similar distributions were found for glutamine synthetase and the ionotropic NMDA receptor subunit NMDAR2. The identification of neurotransmitters and their receptors in the human cochlea has implications for the pharmacotherapy of inner ear diseases.     

Changes in cochlear microphonic response after Y-ray irradiation of the inner ear of the guinea-pig

Summary The effects of ionizing irradiation on the cochlear microphonic response of guinea-pigs were studied. The cochlear microphonics (CM) of both ears were recorded in a total of 36 animals. Recording was carried out by the differential electrode technique on the basal turn of the cochlea. One week prior to recording, the left ear of each animal had been exposed to 35–70 Gy radiation in increments of 5 Gy. Doses of 40 Gy and above led to a reduction in CM response. After doses of 60 Gy or more, no CM response could be detected. Damage most probably occurred in the outer and inner hair cells.     

What drives mechanical amplification in the mammalian cochlea?

The recent report by Peter Dallos and colleagues of the gene and protein responsible for outer hair cell somatic motility (Zheng, Shen, He, Long, Madison, & Dallos, 2000), and the work of James Hudspeth and colleagues demonstrating that vestibular stereocilia are capable of providing power that may boost the vibration of structures within the inner ear (Martin & Hudspeth, 1999), presents the tantalizing possibility that we may not be far away from answering the question what drives mechanical amplification in the mammalian cochlea? This article reviews the evidence for and against each of somatic motility as the motor, and a motor in the hair cell bundle, producing cochlear mechanical amplification. We consider three models based on somatic motility as the motor and two based on a motor in the hair cell bundle. Available evidence supports a hair cell bundle motor in nonmammals but the upper frequency limit of mammalian hearing in general exceeds that of nonmammals, in many cases by an order of magnitude or more. Only time will tell whether an evolutionary dichotomy exists (Manley, Kirk, K?ppl, & Yates, 2001).