Schwann cells genetically modified to express neurotrophins promote spiral ganglion neuron survival in vitro.

The intracochlear infusion of neurotrophic factors via a mini-osmotic pump has been shown to prevent deafness-induced spiral ganglion neuron (SGN) degeneration; however, the use of pumps may increase the incidence of infection within the cochlea, making this technique unsuitable for neurotrophin administration in a clinical setting. Cell- and gene-based therapies are potential therapeutic options. This study investigated whether Schwann cells which were genetically modified to over-express the neurotrophins brain-derived neurotrophic factor (BDNF) or neurotrophin 3 (Ntf3, formerly NT-3) could support SGN survival in an in vitro model of deafness. Co-culture of either BDNF over-expressing Schwann cells or Ntf3 over-expressing Schwann cells with SGNs from early postnatal rats significantly enhanced neuronal survival in comparison to both control Schwann cells and conventional recombinant neurotrophin proteins. Transplantation of neurotrophin over-expressing Schwann cells into the cochlea may provide an alternative means of delivering neurotrophic factors to the deaf cochlea for therapeutic purposes.

Focused electrical stimulation using a single current source.

OBJECTIVE: Cochlear implants, while providing significant benefits to recipients, remain limited due to broad neural activation. Focussed multipolar stimulation (FMP) is an advanced stimulation strategy that uses multiple current sources to produce highly focussed patterns of neural excitation in order to overcome these shortcomings. APPROACH: This report presents single-source multipolar stimulation (SSMPS), a novel form of stimulation based on a single current source and a passive current divider. Compared to conventional FMP with multiple current sources, SSMPS can be implemented as a modular addition to conventional (i.e. single) current source stimulation systems facilitating charge balance within the cochlea. As with FMP, SSMPS requires the determination of a transimpedance matrix to allow for focusing of the stimulation. The first part of this study therefore investigated the effects of varying the probe stimulus (e.g. current level and pulse width) on the measurement of the transimpedance matrix. SSMPS was then studied using in vitro based measurements of voltages at non-stimulated electrodes along an electrode array in normal saline. The voltage reduction with reference to monopolar stimulation was compared to tripolar and common ground stimulation, two clinically established stimulation modes. Finally, a proof of principle in vivo test of SSMPS in a feline model was performed. MAIN RESULTS: A probe stimulus of at least 40 nC is required to reliably measure the transimpedance matrix. In vitro stimulation using SSMPS resulted in a significantly greater voltage reduction compared to monopolar, tripolar and common ground stimulation. Interestingly, matching measurement and stimulation parameters did not lead to an improved focussing performance. Compared to monopolar stimulation, SSMPS resulted in reduced spread of neural activity in the inferior colliculus, albeit with increased thresholds. SIGNIFICANCE: The present study demonstrates that SSMPS successfully limits the broadening of the excitatory field along the electrode array and a subsequent reduction in the spread of neural excitation.

Inosine binds to A3 adenosine receptors and stimulates mast cell degranulation.

We investigated the mechanism by which inosine, a metabolite of adenosine that accumulates to > 1 mM levels in ischemic tissues, triggers mast cell degranulation. Inosine was found to do the following: (a) compete for [125I]N6-aminobenzyladenosine binding to recombinant rat A3 adenosine receptors (A3AR) with an IC50 of 25+/-6 microM; (b) not bind to A1 or A2A ARs; (c) bind to newly identified A3ARs in guinea pig lung (IC50 = 15+/-4 microM); (d) lower cyclic AMP in HEK-293 cells expressing rat A3ARs (ED50 = 12+/-5 microM); (e) stimulate RBL-2H3 rat mast-like cell degranulation (ED50 = 2.3+/-0.9 microM); and (f) cause mast cell-dependent constriction of hamster cheek pouch arterioles that is attenuated by A3AR blockade. Inosine differs from adenosine in not activating A2AARs that dilate vascular smooth muscle and inhibit mast cell degranulation. The A3 selectivity of inosine may explain why it elicits a monophasic arteriolar constrictor response distinct from the multiphasic dilator/constrictor response to adenosine. Nucleoside accumulation and an increase in the ratio of inosine to adenosine may provide a physiologic stimulus for mast cell degranulation in ischemic or inflamed tissues.

Gentamicin Applied to the Oval Window Suppresses Vestibular Function in Guinea Pigs.

Intratympanic gentamicin therapy is widely used clinically to treat the debilitating symptoms of Ménière's disease. Cochleotoxicity is an undesirable potential side effect of the treatment and the risk of hearing loss increases proportionately with gentamicin concentration in the cochlea. It has recently been shown that gentamicin is readily absorbed through the oval window in guinea pigs. The present study uses quantitative functional measures of vestibular and cochlea function to investigate the efficacy of treating the vestibule by applying a small volume of gentamicin onto the stapes footplate in guinea pigs. Vestibular and cochlea function were assessed by recording short latency vestibular evoked potentials in response to linear head acceleration and changes in hearing threshold, respectively, 1 and 2 weeks following treatment. Histopathology was analyzed in the crista ampullaris of the posterior semi-circular canal and utricular macula in the vestibule, and in the basal and second turns of the cochlea. In animals receiving gentamicin on the stapes footplate, vestibular responses were significantly suppressed by 72.7 % 2 weeks after treatment with no significant loss of hearing. This suggests that the vestibule can be treated directly by applying gentamicin onto the stapes footplate.

Perinuclear mRNA localisation by vimentin 3'-untranslated region requires a 100 nucleotide sequence and intermediate filaments.

The role of the vimentin 3'-untranslated region (3'-UTR) in mRNA localisation was studied in cells transfected with a reporter sequence linked to subregions of the 3'-UTR. In situ hybridisation showed that nucleotides 37-137, including a previously identified protein-binding domain, were sufficient to localise transcripts to perinuclear cytoplasm. Transfection of two SW13 cell lines that do and do not express vimentin showed that perinuclear localisation due to either the vimentin or c-myc 3'-UTR requires intermediate filaments. The data suggest that both a specific protein-binding region of the vimentin 3'-UTR and intermediate filaments themselves are required to determine the site of vimentin synthesis.

Hearing, vocalization and the external ear of a marsupial, the northern Quoll, Dasyurus hallucatus.

As part of a continuing study of the development of the marsupial auditory system, auditory brainstem responses (ABR) were recorded and an ABR audiogram was constructed for five female Northern Quolls (Dasyurus hallucatus), which are nocturnal carnivores. The best frequency for hearing lies between 8 and 10 kHz, and at 50 dB SPL there is a range from about 0.5 to 40 kHz. Vocalizations of adult quolls and pouch-young were recorded with a digital audio tape recorder, and the power spectra of representative calls were compared with the ABR audiogram. The common adult vocalizations have most energy at the lower end of the hearing range, whereas frequencies that are dominant in the isolation calls of the pouch-young lie close to the best frequency of hearing. Samples of nocturnal sounds of the habitat of the quoll were also recorded and analyzed. Power spectra have peak energy at frequencies between 2 and 5 kHz, with a smaller contribution above 10 kHz. The spectrum contains relatively little power at the best frequency of hearing. Measurements of the sound pressure level at the external ear canal as a function of stimulus frequency and location in space suggest that the directional amplifying properties of the pinna will operate most effectively on sound frequencies at the upper end of the quoll's hearing range, a region that may be important in prey detection. Comparisons are made with other mammalian nocturnal carnivores and with other marsupials. We speculate that, for nocturnal carnivores, one role of the low-frequency part of the hearing range concerns the recognition of adult conspecifics, the mid-frequency range is important for the detection of pouch-young, and the upper range may be particularly concerned with prey/predator detection.

Distortion product otoacoustic emission and auditory brainstem responses in the echidna (Tachyglossus aculeatus).

The auditory function of four wild-caught echidnas was measured using distortion product otoacoustic emissions (DPOAEs) and auditory brainstem responses (ABRs). Emission audiograms were constructed by finding the stimulus levels required to produce a criterion emission amplitude at a given stimulus frequency. For an emission amplitude of -10 dB SPL, the median "best threshold" was 28 dB SPL, and this minimum threshold occurred between 4 and 8 kHz for all animals. The relative effective range of auditory function was defined by the frequencies at which the audiogram was 30 dB above its best threshold. For the emission audiograms, the median lower-frequency limit was 2.3 kHz, the upper limit was 18.4 kHz, and the effective range was 2.7 octaves. The audiogram as measured by ABR was also found to be strongly "U" shaped with similar low- and high-frequency limits, i.e., from 1.6 to 13.9 kHz, with an effective range of 3.1 octaves. These results suggest that the echidna has a behavioral hearing sensitivity comparable to that of typical therian mammals (e.g., rabbits and gerbils) but with a significantly narrower frequency range. DPOAE responses were also measured in selected animals as a function of the variation of all four stimulus parameters (frequencies and intensities of both stimulus tones). Overall, the measured emission responses establish that the echidna does have a cochlear amplifier, and that it could be the same type as in therian mammals. The amplification mechanism in the echidna, currently unidentified, clearly operates to frequencies above 20 kHz, higher than the hearing function observed in any birds or reptiles but lower than for typical therian mammals. This raises the possibility that at least some aspects of the mammalian cochlear amplifier developed early in evolution, before the divergence of the monotremes (echidna and platypus) from the mainstream therian mammals (marsupials and placentals). In this respect, the presence or absence of outer hair cell electromotility in monotremes would have important consequences for understanding the function and evolution of the vertebrate inner ear.

Scanning electron microscopy of platinum scala tympani electrodes following chronic stimulation in patients.

Platinum (Pt) electrodes from three auditory prostheses (Cochlear Pty Ltd) were examined for evidence of corrosion following implantation periods of up to 1000 days. These devices were used for periods ranging from 1600 to 10,400 h and developed maximum charge densities of 0.257 microC mm-2 geom. per phase. Scanning electron microscopy of the surface of the 66 stimulated electrodes examined showed no evidence of definitive Pt corrosion. Their surface features were essentially identical to control (unstimulated) electrodes. In addition, there was no evidence of any change in the surface morphology of the Silastic carrier adjacent to the stimulating electrodes. These results indicate that Pt is a suitable electrode material for neural prostheses that use relatively large surface area electrodes (0.1-1.0 mm2) and low to moderate charge densities (0.01-0.26 microC mm-2 geom. per phase).

Scanning electron microscopy of chronically stimulated platinum intracochlear electrodes.

Platinum electrodes were examined for evidence of corrosion using a scanning electron microscope (SEM). In vivo electrodes, stimulated using charge-balanced biphasic pulses for periods of up to 2000 h at charge densities of 0.18-0.32 microC mm-2 geom. per phase, were compared with in vitro electrodes stimulated in inorganic saline using similar stimulus parameters, and with in vivo control electrodes. The in vitro stimulated electrodes showed evidence of platinum corrosion at high charge density and aggregate charge injection. Significantly, the in vivo stimulated electrodes showed no evidence of stimulus induced corrosion. Indeed, their surfaces were similar to the in vivo control electrodes. In vitro electrochemical studies have demonstrated that proteins play a significant role in the inhibition of platinum dissolution: the present study has demonstrated an inhibitory effect in vivo. This may be due to the presence of proteins.

Design and fabrication of the banded electrode array.

The banded electrode array is a simple and effective array for multiple-channel cochlear stimulation and meets the necessary design requirements.

LIF potentiates the NT-3-mediated survival of spiral ganglia neurones in vitro.

The survival of auditory neurones depends on the continued supply of trophic factors. Early postnatal spiral ganglion cells (SGC) in a dissociated cell culture were used as a model of auditory innervation to test the trophic factors leukaemia inhibitory factor (LIF) and neurotrophin-3 (NT-3) for their ability, individually or in combination, to promote neuronal survival. The findings suggest that LIF supports neuronal survival in a concentration-dependent manner. Moreover LIF potentiated NT-3-mediated spiral ganglion neuronal survival in a synergistic fashion.

Neonatal sensorineural hearing loss affects synaptic density in the auditory midbrain.

We examined the effect of neonatal sensorineural hearing loss on synaptic density in the central nucleus of the inferior colliculus (ICC) of adult cats to evaluate the role of auditory experience in synaptogenesis. Three groups of animals were used: bilaterally deafened, unilaterally deafened and normal hearing controls. Synaptic density in bilaterally deafened animals was significantly lower than in normal hearing animals. By contrast, there was no significant difference in synaptic density between normal hearing animals and unilaterally deaf animals. These results demonstrate, for the first time, that a sensorineural hearing loss during development can affect synaptogenesis in the auditory midbrain.

Synergy between TGF-beta 3 and NT-3 to promote the survival of spiral ganglia neurones in vitro.

Transforming growth factor-betas (TGF-betas) have been implicated in normal inner ear development and in promoting neuronal survival. Early rat post-natal spiral ganglion cells (SGC) in dissociated cell culture were used as a model of auditory innervation to test the trophic factors TGF-beta3 and neurotrophin-3 (NT-3) for their ability, individually or in combination, to promote neuronal survival. The findings from this study suggest that TGF-beta3 supports neuronal survival in a concentration-dependent manner. Moreover TGF-beta3 and NT-3-potentiated spiral ganglion neuronal survival in a synergistic fashion.

Electrical stimulation of the auditory nerve. III. Response initiation sites and temporal fine structure.

Latency, temporal dispersion and input-output characteristics of auditory nerve fiber responses to electrical pulse trains in normal and chronically deafened cat ears were classified and tentatively associated with sites where activity is initiated. Spikes occurred in one or more of four discrete time ranges whose endpoints overlapped partially. A responses had latencies <0.44 ms, exhibited asymptotic temporal dispersion of 8-12 micros and possessed an average dynamic range of 1.2 dB for 200 pulses/s (pps) pulse trains. They likely originated from central processes of spiral ganglion cells. B(1) and B(2) responses (0.45-0.9 ms, 25-40 micros, 1.9 dB) likely stemmed from activity at myelinated and unmyelinated peripheral processes, respectively. C100 micros) likely originated from direct stimulation of inner hair cells, and D8 dB) arose from propagating traveling waves possibly caused by electrically induced motion of outer hair cells. C and D responses were recorded only in acoustically responsive ears. Mean latencies of spikes in all time ranges usually decreased with intensity, and activity at two or even three discrete latencies was often observed in the same spike train. Latency shifts from one discrete time range to another often occurred as intensity increased. Some shifts could be attributed to responses to the opposite-polarity phase of the biphasic pulse. In these cases, temporal dispersion and dynamic range were approximately equal for activity at each latency. A second type of latency shift was also often observed, in which responses at each latency exhibited dissimilar temporal dispersion and dynamic range. This behavior was attributed to a centralward shift in the spike initiation site and it occurred for monophasic as well as biphasic signals. Several fibers exhibited dual latency activity with a 40-90 micros time difference between response peaks. This may have stemmed from spike initiation at nodes on either side of the cell body. Increasing the stimulus pulse rate to 800-1000 pps produced small increases in temporal dispersion and proportionate increases in asymptotic discharge rate and dynamic range, but thresholds did not improve and slopes of rate-intensity functions (in spikes/s/dB) did not change. Responses to high-rate stimuli also exhibited discrete latency increases when discharge rates exceeded 300-400 spikes/s. Spike by spike latencies in these cases depended strongly on the discharge history. Implications for high-rate speech processing strategies are discussed.

Progressive ototoxicity of neomycin monitored using derived brainstem response audiometry.

Progressive hearing loss following the systemic administration of neomycin was investigated using derived brainstem response audiometry. Cats were given three to five times the maximum recommended clinical dose of neomycin over a period of 10 days. Their hearing was monitored prior to and during the administration of the drug, and periodically following its completion. The results of this study showed that the induced hearing loss generally proceeded from high to low frequencies as an advancing lesion, with regions apical to the lesion functioning normally. Although considerable variability in response to the drug existed among animals, the evoked responses from both ears of each animal showed close bilateral symmetry during the deafening process. Furthermore, the present results highlight the long-term ongoing ototoxicity associated with neomycin, and the importance of monitoring high frequencies for initial signs of an aminoglycoside induced hearing loss.

Electrical stimulation of the auditory nerve. I. Correlation of physiological responses with cochlear status.

The purpose of the present study was to evaluate evoked potential and single fibre responses to biphasic current pulses in animals with varying degrees of cochlear pathology, and to correlate any differences in the physiological response with status of the auditory nerve. Six cats, whose cochleae ranged from normal to a severe neural loss (< 5% spiral ganglion survival), were used. Morphology of the electrically evoked auditory brainstem response (EABR) was similar across all animals, although electrophonic responses were only observed from the normal animal. In animals with extensive neural pathology, EABR thresholds were elevated and response amplitudes throughout the dynamic range were moderately reduced. Analysis of single VIIIth nerve fibre responses were based on 207 neurons. Spontaneous discharge rates among fibres depended on hearing status, with the majority of fibres recorded from deafened animals exhibiting little or no spontaneous activity. Electrical stimulation produced a monotonic increase in discharge rate, and a systematic reduction in response latency and temporal jitter as a function of stimulus intensity for all fibres examined. Short-duration current pulses elicited a highly synchronous response (latency < 0.7 ms), with a less well synchronized response sometimes present (0.7-1.1 ms). There were, however, a number of significant differences between responses from normal and deafened cochleae. Electrophonic activity was only present in recordings from the normal animal, while mean threshold, dynamic range and latency of the direct electrical response varied with cochlear pathology. Differences in the ability of fibres to follow high stimulation rates were also observed; while neurons from the normal cochlea were capable of 100% entrainment at high rates (600-800 pulses per second (pps)), fibres recorded from deafened animals were often not capable of such entrainment at rates above 400 pps. Finally, a number of fibres in deafened animals showed evidence of 'bursting', in which responses rapidly alternated between high entrainment and periods of complete inactivity. This bursting pattern was presumably associated with degenerating auditory nerve fibres, since it was not recorded from the normal animal. The present study has shown that the pathological response of the cochlea following a sensorineural hearing loss can lead to a number of significant changes in the patterns of neural activity evoked via electrical stimulation. Knowledge of the extent of these changes have important implications for the clinical application of cochlear implants.

Onset of ototoxicity in the cat is related to onset of auditory function.

Cats are altricial mammals; they are born deaf and undergo rapid maturation of the auditory periphery late in the first and throughout the 2nd week of life. Previous studies, using multiple aminoglycoside administration over several days or weeks, have indicated that there is a reduction in the degree of ototoxicity in young animals provided the drug is administered prior to the onset of auditory function. In order to provide a more precise relationship between the degree of ototoxicity and auditory development, we used a single administration of Kanamycin (KA) and the loop diuretic ethacrynic acid (EA), as the co-administration of these drugs is known to produce a rapid and profound hearing loss in adult animals. Thirty kittens were administered with KA and EA at ages that varied from 2 to 16 days after birth (DAB) using a fixed dose per kilogram body weight sufficient to profoundly deafen adult animals. All animals made an uneventful recovery from the procedure. At 26 DAB, tone-pip-evoked auditory brainstem responses (ABR) were recorded from each animal in order to establish the extent of the hearing loss. The degree of hearing loss was compared with normal ABR audiograms recorded from 6 age-matched control animals. All animals treated with KA/EA at 9 DAB or older had a profound hearing loss similar to adult animals. Animals treated between 2 and 8 DAB exhibited severe high-frequency hearing losses. The extent of the loss was correlated with age (r = 0.63) and body weight (r = 0.72) such that hearing loss tended to spread towards lower frequencies as age and/or weight increased. All animals exhibited bilaterally symmetrical hearing losses which remained relatively stable over monitoring periods of up to 6 months following the drug treatment. These findings imply that the onset of ototoxicity is related, at least in part, to the onset of auditory function in the kitten. The rapid onset of deafness following this procedure makes it a useful technique in the study of both ototoxicity and cochlear development.

Reduction in excitability of the auditory nerve following electrical stimulation at high stimulus rates: V. Effects of electrode surface area.

High rate intracochlear electrical stimulation at high intensities can induce significant reductions in the excitability of the auditory nerve as measured by a decrement in the amplitude of the electrically evoked auditory brainstem response (EABR). Such changes are primarily associated with stimulus induced neuronal activity, although direct current (DC) can also contribute. We examined the extent of stimulus induced change in auditory nerve excitability using large surface area platinum electrodes ('high-Q' electrodes). These electrodes have a surface area approximately 70 times greater than standard Pt electrodes of the same geometric area, resulting in lower DC and charge density (charge/electrode surface area) for a common stimulus. Guinea pigs were bilaterally implanted with either high-Q or standard Pt electrodes, and unilaterally stimulated for 2 h using stimulus intensities of 12 dB or 20-30 dB above EABR threshold (0.34 microC/phase) at stimulus rates of 200, 400, or 1000 pulses per second (pps). EABRs were recorded before and following the acute stimulation. While there were significant reductions in EABR amplitudes and elevated EABR thresholds following stimulation at 12 dB above threshold using 400 and 1000 pps delivered to standard Pt electrodes, there were fewer or no significant changes in the post-stimulus EABR amplitude and threshold using high-Q electrodes under equivalent stimulus conditions. At a higher stimulus intensity (20-30 dB above EABR threshold), no reduction in EABR amplitude was observed at 200 pps for both stimulating electrodes. However, EABRs were reduced significantly at 400 and 1000 pps. There was significantly greater EABR recovery following stimulation using high-Q electrodes compared with standard Pt electrodes at 400 (P<0.05) and 1000 pps (P<0.05). These data indicate that large surface area electrodes can significantly reduce stimulus induced changes in auditory nerve excitability, and may therefore have important clinical application.

Electrical stimulation of the auditory nerve: II. Effect of stimulus waveshape on single fibre response properties.

To investigate the generation of action potentials by electrical stimulation we studied the response of auditory nerve fibres (ANFs) to a variety of stimulus waveforms. Current pulses were presented to longitudinal bipolar scala tympani electrodes implanted in normal and deafened cochleae. Capacitively coupled monophasic current pulses evoked single ANF responses that were more sensitive to one phase (the 'excitatory' phase) than the other. Anodic pulses produced a significantly shorter mean latency compared with cathodic pulses, indicating that their site for spike initiation is located more centrally along the ANF. The fine temporal structure of ANF responses to biphasic pulses appeared similar to that evoked by monophasic pulses. An excitatory monophasic pulse evoked a significantly lower threshold than a biphasic current pulse having the same polarity and duration leading phase, i.e. the addition of a second phase leads to an increase in threshold. Increasing the temporal separation of the two phases of a biphasic pulse resulted in a moderate reduction in threshold which approached that of an excitatory monophasic pulse for interphase gaps > 100 micros. Greater threshold reductions were observed with narrower current pulses. There was a systematic reduction in threshold with increasing pulse width for biphasic current pulses, reflecting the general charge-dependent properties of ANFs for narrow pulse widths. Chopped biphasic current pulses, which uniformly delivered multiple packets of charge (2 x 30 micros, 3 x 20 micros or 6 x 10 micros) with the same polarity over a 120 micros period, followed by a similar series in the reverse polarity, demonstrated the ability of the neural membrane to integrate sub-threshold packets of charge to achieve depolarisation. Moreover, thresholds for these current pulses were approximately 1.5 dB lower than 60 micros/phase biphasic current pulses with no interphase gap. Finally, stimulation using charge-balanced triphasic and asymmetric current pulses produced systematic changes in threshold and latency consistent with the charge-dependent properties of ANFs. These findings provide insight into the mechanisms underlying the generation of action potentials using electrical stimuli. Moreover, a number of these novel stimuli may have potential clinical application.

Sensorineural hearing loss during development: morphological and physiological response of the cochlea and auditory brainstem.

We have investigated the effects of sensorineural hearing loss on the cochlea and central auditory system of profoundly deafened cats. Seventeen adult cats were used: four had normal hearing; 12 were deafened neonatally for periods of < 2.5 years (five bilaterally, seven unilaterally); and one animal had a long-term (approximately 8 years) profound bilateral hearing loss. Bipolar scala tympani stimulating electrodes were bilaterally implanted in each animal, and electrically evoked auditory brainstem responses (EABRs) were recorded in an acute study to evaluate the basic physiologic response properties of the deafened auditory pathway. The cochleae and cochlear nuclei (CN) of each animal were examined with light microscopy. Spiral ganglion cell density in neonatally deafened cochleae was 17% of normal, and only 1.5% of normal in the long-term deaf animal. There was a 46% reduction in total CN volume in neonatally deafened animals compared to normal, and a 60% reduction in the long-term deaf animal. Neural density in the anteroventral CN of bilaterally deafened animals was 37% higher than normal; 44% higher in the long-term deaf animal. Significantly, however, we saw no evidence of a loss of neurones within the anteroventral CN in any deafened animal. There was a significant increase in EABR threshold and wave IV latency in the deafened animals, and a significant decrease in response amplitude and input/output function gradient. Again, these changes were more extensive in the long-term deaf animal. These data show that a sensorineural hearing loss can evoke significant morphological and physiological changes within the cochlea and auditory brainstem, and these changes become greater with duration of deafness. It remains to be seen whether these changes can be reversed following the introduction of afferent activity via chronic electrical stimulation of the auditory nerve.