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Objective: To verify the usefulness of short-latency vestibular responses evoked by a combination of round window electrical stimulation and sinusoidal rotation (electrovestibular brainstem responses; EVBRs) as a new monitoring tool of the vestibular function in animal experiments., Methods: EVBRs were obtained before, during, and after treatment with aminoglycosides, along with compound action potential (CAP) audiograms. The changes in EVBRs were compared with morphological changes observed by scanning electron microscopy., Results: EVBR amplitudes did not change in the group of guinea pigs treated with amikacin, but markedly decreased in the streptomycin and gentamicin- treated groups. CAP audiograms indicated a significant threshold elevation in the amikacin group, a moderate elevation in the gentamicin group, and no change in the streptomycin group. Under scanning electron microscopy, the loss of the sensory hair cells observed in the cristae ampullares was slight to moderate in the amikacin group, moderate to severe in the streptomycin group, and severe in the gentamicin group., Conclusion: EVBRs reflect overall pathological changes undergone by vestibular hair cells, and support the vestibular specificity of EVBRs.

The magnitudes of suppression of the click-evoked compound action potential of the auditory nerve (CAP), transient click-evoked otoacoustic emissions (TEOAEs) and ensemble background activity of the auditory nerve (EBA), elicited by contralateral acoustical stimulation, were compared in awake or lightly sedated guinea pigs. The contralateral ear was stimulated either by continuous broad-band noise or by low-pass or high-pass noise (intensity 41-62 dB SPL) with cut-off frequencies of 2, 8 and 12 kHz. The maximal suppression of TEOAEs was achieved by contralateral noise containing mainly low frequencies, whereas for suppression of the CAP it was necessary for middle frequencies to be present in the contralateral noise (less than 8 kHz). In contrast to this, EBA was suppressed mainly by high-frequency noise (higher than 8 kHz) whereas low- and middle-frequency noise was ineffective in suppressing EBA. Evaluation of the root mean square voltage of the EBA (filtered in frequency range 0.75-1.25 kHz) enabled the evaluation of fast and slow components of olivocochlear activation. Both fast and slow effects were proportionally suppressed by individual types of contralateral stimulation. The mechanisms of TEOAEs and CAP generation has been confirmed in many earlier studies, but the origin of EBA has yet to be fully elucidated. The obtained data support the hypothesis that a large part of EBA is formed by spontaneous activity of high-frequency-tuned auditory nerve fibres. Suppression of the EBA magnitude during contralateral stimulation may be caused either by a reduced spontaneous firing rate or by a decrease in possible synchronised neuronal firing.

Contralateral broadband noise (BBN) elevates ipsilateral auditory thresholds (central masking) and reduces the amplitude of ipsilateral brainstem auditory evoked potentials (BAEPs). Binaural interactions are complex psychophysical phenomena, but binaural interaction components are easily extracted from BAEPs to monaural versus binaural click stimulation. However, contralateral, or binaural, acoustical stimulation is known to activate simultaneously the crossed and uncrossed medial olivocochlear (MOC) efferent systems and decrease activity in both cochleas. Particularly, contralateral BBN stimulation suppresses in part ipsilateral peripheral activity. What is the role of such contralaterally induced peripheral suppression in the overall changes in central BAEPs observed during contralateral masking or binaural stimulation? Compound action potentials (CAPs) of the auditory nerve and BAEPs were recorded simultaneously in awake guinea pigs from electrodes chronically implanted on the round window of the cochlea and the surface of the brain. Peripheral and central measures of contralateral masking and binaural interactions were obtained from responses to monaural or binaural clicks, with or without contralateral BBN, recorded before, during, and after the reversible blockade of the MOC function following a single intramuscular injection of gentamicin. Contralateral BBN effectively reduced the amplitudes of CAP and of all BAEP peaks. CAP to ipsilateral click did not, however, change significantly from monaural to binaural click stimulation; still, normal binaural interaction components developed in the BAEPs. When the medial efferent function was blocked by gentamicin, the normal contralateral BBN suppression of CAP and of the earliest BAEP peak was lost; however, the later BAEP peaks were suppressed by contralateral BBN as before gentamicin, and the central binaural interaction components were unchanged. In these experimental conditions, the MOC efferent system seems to play little role in centrally recorded contralateral masking and binaural interactions.

The purpose of this study was to evaluate the average discharge rate of all fibres in the whole auditory nerve (R(wn)) when a broad-band noise with steady-state effects is applied to the ear. We assessed the R(wn) parameter by detecting the state of refractoriness of the nerve during noise stimulation using an electric stimulus (ES) as a probe. The technique, applied in awake pre-implanted guinea pigs (Charlet de Sauvage et al., 1994), made it possible to obtain electro-acoustic responses (EARs), from which an estimate of the R(wn) parameter could be deduced. Negative current pulses of 100 micros duration, each followed by an identical pulse of positive polarity for charge balance, were applied between round window and indifferent vertex electrodes at intervals of 160 ms. The 120 ms wide-band noise masker started 92 ms before every other negative ES. The signal on the stimulating electrodes was averaged over a 5.12 ms window in synchrony with the negative pulse. EARs were obtained by alternately subtracting recordings during noise from those during silence. The R(wn) parameter was determined by comparing experimental and computed EAR patterns. For this purpose, a model of unit response incorporating changes in amplitude and conduction velocity during the relative refractory period was designed. The recovery function of the firing probability in response to ES was evaluated. Fibres were classified in different categories according to their background discharge rates. The probability of response of single fibres to ES in each category was calculated on the basis of their interval histograms during silence and noise. Individual spikes were combined accordingly to obtain the computed EAR waveform. R(wn) was determined by adjusting the EAR amplitude of the model in relation to that of the experimental EAR. R(wn) generally increases in a linear fashion with respect to noise intensity expressed in dB, thus following the increase in loudness perception estimated by Weber's law. At the highest noise levels, R(wn) tends to saturate. The estimated saturation rate was found to be about 380 spikes/s.

The first detectable effect on the auditory system after a single high-dose injection of an aminoglycosidic antibiotic (AA) like gentamicin (GM) is the reversible blockade of medial efferent function, probably via blockade of calcium channels at the base of the outer hair cells (OHC). The kinetics of this effect are compatible with that of the molecule in perilymph. In the course of chronic treatment with lower doses, however, ototoxicity develops only after several days of treatment. Still GM can be observed inside the OHCs as soon as 24 hours after the first injection, and will be still present in some OHCs as long as 11 months after a chronic, nonototoxic 6-day treatment. In vitro, the short-term viability of isolated OHCs is not affected by exposure to AAs, but their transduction channels and their response to acetylcholine are reversibly blocked. However, developing organs of Corti in culture are highly and rapidly affected by exposure to AAs. Yet during direct intracochlear perilymphatic perfusion of GM, 2-mM solutions are not ototoxic, and with perfusion with a 20-mM solution ototoxicity develops only after several days of perfusion. From these various observations one can describe some aspects of the mechanisms of ototoxicity of AAs, from their access to perilymph and endolymph, to penetration in the hair cells, likely via endocytosis at their apical pole, and intracellular cytotoxic events.

Free radical generation is increasingly implicated in a variety of pathological processes, including drug toxicity. Recently, a number of studies have demonstrated the ability of gentamicin to facilitate the generation of radical species both in vivo and in vitro, which suggests that this process plays an important role in aminoglycoside-induced ototoxicity. Free radical scavengers are compounds capable of inactivating free radicals, thereby attenuating their tissue damaging capacity. In this study we have determined the ability of the powerful free radical scavenger alpha-lipoic acid (100 mg/kg/day) to attenuate the cochlear damage induced by a highly ototoxic regimen of the aminoglycoside amikacin (450 mg/kg/day, i.m.). Experiments were carried out on pigmented guinea pigs initially weighing 200-250 g. Changes in cochlear function were characterized as shifts in compound action potential (CAP) thresholds, estimated every 5 days, by use of chronic indwelling electrodes implanted at the round window, vertex, and contralateral mastoid. Results showed that animals receiving alpha-lipoic acid in combination with amikacin demonstrated a significantly less severe elevation in CAP thresholds compared with animals receiving amikacin alone (P < 0.001; t-test). These results provide further evidence of the recently reported intrinsic role of free radical generation in aminoglycoside ototoxicity, and highlight a potential clinical therapeutic use of alpha-lipoic acid in the management of patients undergoing aminoglycoside treatment.

Auditory brainstem responses (ABRs) to click stimuli have been compared in young (21-day-old), adult (6-month-old), and old (18-month-old) rats fed a normal (Arachid-Colza) or an alpha-linolenic acid deficient (Arachid only) diet. Wave I amplitude and latency did not show any significant change with either age or diet. However, wave III showed a progressive decrease in amplitude and latency from young to adult and from adult to old rats having a normal diet. With alpha-linolenic acid deficiency, wave III amplitude and latency values decreased faster than in the normal diet control groups. Although final values in the old groups with the two diets were similar, with alpha-linolenic acid deficiency values for wave III decreased to this final level in the adult group. These data indicate that the central auditory nervous system ages faster, or earlier, with a fatty acid deficiency.

Objectives: To establish that susceptance-conductance tympanograms at a probe-tone frequency of 2 kHz reflects the status of the annular ligament (AL) and through it of the cochlea., Methods: Experimental study in 5 chinchillas and 22 guinea pigs. Six validating experiments were used: blockages of the stapes and of the round window membrane (RWM), fistula of the RWM, fluid removal from the cochlea, injection of saline in the scala tympani (ST) and acoustic trauma (AT). Quantitative data (mean values of Y226, FR, Y2000, G2000 and B2000) and shape of the curves were analyzed before and immediately after lesions were done., Results: Guinea pig was the most convenient provided bulla was vented and the same tip was used along the experiments. Only the shape of the curves are discriminant: 1/a supplementary sharp peak, centered around negative pressures, is observed in Y/G tympanograms in every case of RWM fistulas and in some case of AT. 2/injection of saline into ST induces immediate and reproducible Y2000, G2000, et B2000 curves modifications. 3/RWM and stapes blockages provoke foreseeable stiffening and sharpening of the tympanograms at 2 kHz. 4/on the contrary, fluid removal from the cochlea induces multiple peaks curves., Conclusions: Experimentally-induced modifications at the AL either direct (stapes blockage) or indirect by AT or decrease/increase of pressure load at the cochlear interface at the footplate result in noticeable, constant, reproducible changes of curves registered at 2 kHz. The stapes behaves both as the plotter of the curves and the interpreter of the inner ear pressure.

The 2f1-f2 distorsion product (DPOAE) was measured in conjunction with contralateral noise to evaluate the short-term effectiveness of the olivocochlear efferents in guinea pigs (GPs). An attenuation effect was observed predominantly between 1 and 6 kHz when primary tones were set at 65 dB SPL (contralateral noise of 55 dB SPL). Subsequently, GPs were exposed to a 2 kHz tone of 87 dB SPL for 40 min, using DPOAEs as an estimate of cochlear sensitivity. The response of the cochlea appeared variable. In order to investigate whether effectiveness of efferents plays a role in temporary threshold shift (TTS), the responses of the cochlea to overstimulation were classified into three groups: i) clear cochlear change with complete recovery or actual TTS (group A1); ii) clear cochlear change with incomplete recovery (group A2); iii) mild or no change in cochlear function (group B). No relationship was found between the attenuation effect measured before noise overexposure and the susceptibility to TTS. Animals with a significant attenuation effect could fit into any of the three groups. In addition, the recovery from loud sound exposure was not paralleled with the changes occurring over time in the attenuation effect. Therefore, the conclusion that short-term effectiveness of medial efferents does not predict susceptibility to TTS in the GP is suggested.

Recently our laboratory has demonstrated, in the guinea pig (GP), that an intramuscular (i.m.) injection of a high dose of gentamicin (GM) (150 mg/kg), can reversibly block the contralateral efferent suppression of ipsilateral cochlear activity. The aims of the present study were: (1) to investigate this effect with lower doses of GM; and (2) to find out whether this effect could constitute an anticipatory sign of ototoxicity during a chronic GM treatment (60 mg/kg i.m., 10 days). The function of the medial olivocochlear efferent system (MOES) was tested by recording the VIIIth nerve ensemble background activity (EBA) without and with contralateral low level (55 dB SPL) broadband noise stimulation. The results show a dose-dependent effect of GM on contralateral suppression, as the dose of 120 mg/kg induced a smaller blockade of the MOES, compared to 150 mg/kg, and no blockade was observed with lower doses. During the ten-day treatment no significant changes in the EBA without acoustic stimulation, nor in contralateral efferent suppression were detected. GPs monitored over several weeks after the treatment showed progressive reduction of the EBA without contralateral stimulation parallel to reduced suppression coefficients of the EBA, and CAP threshold elevations, denoting impaired cochlear function. Thus, this study demonstrated that a chronic treatment with 60 mg/kg of GM, although ototoxic, does not affect the contralateral efferent suppression, at least before the development of ototoxicity.

Gentamicin (GM) has been shown to reversibly reduce the ability of contralateral noise to suppress ipsilateral cochlear activity, in a dose-dependent manner. However, during chronic administration of lower doses (60 mg/kg) the involvement of medial efferents could not be demonstrated. The purposes of the present study were to determine whether other aminoglycosides would display the same acute effects as GM and whether there was any correlation between their specificity and degree of cochlear and vestibular toxicity and their potency of blockade of the medial efferent system. Thus, we observed changes in ipsilateral ensemble background activity (EBA) of the VIIIth nerve without and with contralateral low level (55 dB SPL) broadband noise stimulation, in awake guinea pigs (GPs), before and after one single high-dose intramuscular injection of different aminoglycoside antibiotics (AAs) (gentamicin, amikacin, neomycin, netilmicin, streptomycin, tobramycin). For comparison, the effects of strychnine, a known antagonist of the efferent transmission and of cisplatin, an antineoplastic agent with cochleotoxic properties were also studied. Netilmicin displayed blocking properties similar to GM, although less pronounced, while amikacin and neomycin had no effect on medial efferent function. With tobramycin and streptomycin a decrease in suppression was usually associated with a reduction of the EBA measured without acoustic stimulation. However, with cisplatin, suppression was still effective when EBA was severely decreased. We could not observe specific effects of strychnine on medial efferent function. In conclusion, no correlation was found between specificity and degree of AA ototoxicity and their action on the medial efferent system.

The function of the medial olivocochlear efferent system was observed in awake guinea pigs by recording, in the absence of ipsilateral external acoustic stimulation, the ensemble background activity (EBA) of the VIIIth nerve from an electrode chronically implanted on the round window of one ear. The EBA was measured by calculating the power value of the round window signal in the 0.5- to 2.5-kHz band after digital or analog (active) filtering. This EBA was compared with and without the addition of a low-level broadband noise to the opposite ear. The contralateral broadband noise (CLBN, 55 dB SPL) induced, via the efferent system, a decrease (suppression) of this EBA. With the use of noise bursts of different durations, two components in this suppression could be observed. After the onset of a 1-s CLBN, the power value of the EBA decreased rapidly by 38.0 +/- 4.2% (mean +/- SD, n = 3), with a latency of <10 ms and a decay time constant of 13.1 +/- 1.0 ms (fast effect). At the offset of the 1-s CLBN, EBA came back to prestimulation values with a similar latency and a time constant of 15.5 +/- 2.9 ms. During longer CLBN stimulation (>/=1 min), EBA presented, after the fast decrease, an additional, slower decrease of 15.6 +/- 3.1%, with a delay of 9.8 +/- 1.3 s and a decay time constant of 16.1 +/- 5.0 s (n = 12, slow effect), and then remained remarkably constant for as long as observed, i.e., >2 h (steady state). The average global suppression was thus up to 47.8 +/- 5.8% of the basal, pre-CLBN-stimulation EBA value. At the offset of the CLBN, EBA returned to pre-CLBN level with fast and slow phases, with, for the slow phase, no delay and a time constant of 32.1 +/- 8.1 s. Fast and slow changes in EBA power values were observed after a single injection of gentamicin (GM) at different doses (150, 200, and 250 mg/kg). At 150 and 200 mg/kg, GM progressively and reversibly blocked the rapid effect, but the slow component of the efferent medial suppression remained remarkably unchanged. However, at higher doses both the fast and slow suppressions were totally yet still reversibly blocked. These observations indicate that the medial olivocochlear efferent system exerts sustained influences on outer hair cells and that this effect develops in two different steps that may have different basic cellular mechanisms.

The bioelectrical activity of the cochlea, without any ipsilateral acoustic stimulation, was recorded in awake guinea pigs (GPs) between electrodes chronically implanted at the round window (RW) and the skull. Measuring its power in the band centered around 1.0 kHz (0.5-2.5 kHz) provided an indirect measure of the ensemble background (EBA) activity of the eighth nerve. Contralateral white-noise (CLWN) stimulation reduced this EBA, presumably by activation of medial olivocochlear fibers. The aim of the investigation was to validate measurements of EBA and of its contralateral suppression in order to study the medial efferent function. The first goal was to find the best conditions for recording the EBA in the absence of ipsilateral stimulation and for studying its suppression by contralateral acoustic stimulation, which implies that no noise was generated by the experimental animal. Thus recordings were compared in normal, awake GPs and in GPs under sedation with xylazine, anesthetized with a combination of xylazine and ketamine, and with and without temperature regulation. In order to monitor the effects of sedation and anesthesia, the recordings were analyzed not only in the 0.5- to 2.5-kHz frequency band but also in the other frequency bands, 5-50 Hz, 50-150 Hz, and 150-500 Hz, which presumably include general central and neuromuscular contributions. The results show that sedation with xylazine accompanied by regulation of body temperature does not affect the EBA value nor its contralateral suppression. Nevertheless, anesthesia should be avoided, even with control of body temperature. The second goal of this study was to identify the specific cochlear contribution to the raw RW signal. Thus recordings were performed in normal and deafened animals and analyzed in the frequency band 0.5-2.5 kHz and also in the other frequency bands of 5-50 Hz, 50-150 Hz, and 150-500 Hz. The results indicate that most of the cochlear activity lies in the frequency band 0.5-2.5 kHz, with also some minor contribution coming from the 150- to 500-Hz band. Analysis and comparison of power values in the different conditions indicate that specific cochlear EBA power was about 60 microV2. From a commonly accepted mean background discharge rate of 50 spikes/s (sp/s), the EBA power without CLWN should have been around 4.4 microV2 if the fibers' activity was random. This difference suggests that there is probably some degree of synchrony between individual fibers. There was a reduction of approximately 45% during CLWN stimulation. This suppression might correspond to a reduction in both discharge rate and synchrony of the fibers.

In the electrical stimulation (ES) of auditory pathways, the type of stimulus and the electrode/tissue interface are critical parameters for the safety and efficacy of the protocol. In this study the influence of alternate pulses, applied between round window and vertex electrodes in chronically implanted guinea pigs, and maintained during 1 and 25 daily periods of 2 h (short-term and long-term experiments, respectively), was investigated. ES consisted of monophasic current pulses of +/- 70 microA and 300 (micro)s in duration at a rate of 167/s, with alternate polarity. Compound Action Potential (CAP) audiograms, amplitudes and latencies of click-evoked CAPs, amplitudes and latencies of electrically-evoked auditory responses (EARs), and electrode impedances, were measured periodically outside or during the ES periods. Short-term ES induced no change in CAP thresholds, amplitude and latency in response to clicks at 80 dB above normal threshold, but induced a slight latency increase and amplitude decrease of the EAR, correlated with an exponential decay of the electrode impedance. On a long-term basis, CAP audiograms and latencies did not change significantly, whereas CAP amplitudes and electrode impedances increased, in correlation with each other. In control guinea pigs receiving no ES, the same CAP amplitude and impedance changes were observed over the same long-term period. The EAR and CAP changes can be explained by a variation of the electrical impedance of the electrode/tissue interface. This is possibly due to a change in electrolytes around the electrode under the influence of the ES for the short-term variation, and to an electrode encapsulation by fibrous tissue independent of the ES for the long-term change. In itself, and under the conditions of this experiment, the ES demonstrated no adverse effects on the auditory function and can be safely used for inner-ear exploration.

Using the guinea pig middle ear model, we assessed decalcified, lyophylized, sterile heterotopic porcine ossicular xeno-implants based on a histology (optic and electron scan microscope) and immunologic (immunofluorscence) methods. Implants were placed in the middle ear and others in the dorsal subcutaneous area. Allo-implants were compared as controls. Implants were placed in the middle ear in 54 animals and skin implants in 14. Under the influence of BMP, the implant ossified in all cases in the middle ear. Intense immune recruitment was not observed. Inversely, there was a mononuclear infiltration reaction to the skin implants with formation of a fibrous capsule, immunoglobulin and complement influx and consequently sequestration. The allo-implants were partially reossified. These findings confirm the value of decalcification with hydrochloric acid for BMP induction, independent of species and the failure of attempted immune despecification. Implant outcome is not dependent on its antigen load, which is high compared with its weight, but on the site of implantation. The middle ear appears to be a privileged site of implantation.

The influence of xylazine on the amplitude, latency and waveform of VIIIth nerve compound action potential (CAP) and cochlear microphonic (CM) in response to clicks at 95 dB SPL in normal awake preimplanted guinea pigs was investigated. The animals' temperature was monitored but no thermoregulation was exerted, except in one control experiment. Following a 0.2 ml injection of xylazine, CM showed minor variations while CAP audiograms for tone pips between 0.5 and 25 kHz remained normal. However, a progressive decrease in temperature and a strongly correlated increase in CAP amplitude and in N1 and N2 latencies were noticed. For peak N1 the changes were equivalent to linear amplitude and time expansions, and could be reproduced through CAP synthesis with convolution methods using time expanded unit response model and firing density functions. All changes were maximal after 2 h of sedation and recovered within approximately another 2 h. Whereas xylazine is known to induce hypothermia, all the changes disappeared if the animal was thermoregulated. Therefore the changes are interpreted as a result of hypothermia. The mechanism of N1 latency lengthening and increase in amplitude during hypothermia can be understood as a simultaneous increase in spike duration, hair cell/nerve synaptic delay and postsynaptic time constant. This hypothesis yielded a theoretical temperature coefficient for N1 latency (-52 microseconds/degree C) matching that measured experimentally (-55 microseconds/degree C). When compared with peak N1, peak N2 appeared relatively more expanded. Arguments about the origin of N2 are discussed.

The physiology of the medial efferent olivocochlear system involves suppressive interactions of contralateral sounds on ipsilateral sound-evoked responses, but its role is largely unknown to date. Medial efferents act at the level of cochlear outer hair cells via cholinergic synapses and might affect their mechanical activity, thereby modulating auditory sensitivity. The aim of the present work was to obtain noninvasive measurements of distortion-product otoacoustic emissions (DPOEs), which reflect outer hair cell function, in order to establish the characteristics of medial efferent-induced suppression in awake, restrained guinea pigs. A clear suppression of DPOEs was induced by continuous contralateral white noise presented at 20-70 dB SPL, in the absence of any confounding effect of anesthesia, middle-ear muscles, or acoustic cross talk. Recently, acute injection of a high dose of the aminoglycoside antibiotic gentamicin (150 mg/kg) was reported to alter the suppressive effect of contralateral noise on eighth nerve-compound action potentials, presumably by blocking efferent synapses to outer hair cells. This hypothesis was confirmed with DPOEs for which a single injection of gentamicin at the same dose abolished suppression after about 1-2 h, whereas no change in basal levels was observed. Complete recovery was obtained after 48 h. This experiment may provide an easy, noninvasive tool for studying auditory function with and without functioning efferents.

The distribution of amikacin (AK), an exclusive cochleo-toxic aminoglycosidic antibiotic (AA), and of gentamicin (GM), which is both cochleo- and vestibulo-toxic, has been studied in cochlear and vestibular hair cells. Guinea pigs were treated during six days with one daily injection of AK (450 mg/kg/day) or GM (60 mg/kg/day). AAs were detected, using immunocytochemical technique with scanning laser confocal microscopy, in isolated cells from guinea pigs sacrificed from 2 to 30 days after the end of the treatments. Results demonstrate a rapid uptake (as soon as after 2-day treatment) of both AAs by cochlear and vestibular hair cells and a very slow clearance. Particularly GM and AK are detected in type I and type II hair cells of the utricles and cristae ampullaris. The presence of these two molecules with different toxic potentialities towards cochlear and vestibular hair cells indicates that the selective ototoxicity of aminoglycosides cannot be explained simply on the basis of particular uptake and accumulation in the different sensory hair cells.

Responses to electrical stimulation of the ear applied between round-window and vertex electrodes were recorded in awake guinea-pigs from the same electrodes or from separate vertex/mastoid subdermal needle electrodes. They were averaged during opposite phases of sinusoidal rotation or before and after constant velocity rotation. In both cases the responses were subtracted from each other and yielded differential per- or post-rotatory "electrovestibular" responses. For comparison, responses were also recorded in the same animals and conditions of electrical stimulation during silence and during silence and during presentation of a broad-band noise. The difference yielded "electroacoustic" responses. In round-window records, electrovestibular and electroacoustic responses presented typical compound nerve action potential patterns. Electrovestibular responses could be recorded for head angular velocities as low as 3 degrees sec-1 at 0.1 Hz. Response amplitude showed a logarithmic relation to head velocity. Changes in amplitude, as a function of time after rotation, were comparable to those reported for vestibular nerve fibre responses. In vertex/mastoid records, electroacoustic responses presented a sequence of peaks similar to the click-evoked auditory brain-stem responses, and electrovestibular responses presented two peaks, presumably representing contributions of central vestibular structures. Such "electrovestibulography" permits the study of an individual ear and makes available to the investigator a large range of vestibular stimulation conditions.

Oral administration of glycerol at 0.5 g/kg and 1.0 g/kg body weight in the guinea pig with experimentally induced hydrops provoked no obvious modifications of the CAP audiogram within a 3-h recording session. Daily administration of glycerol at the same dose level during a 4-month period did not modify the deterioration of CAP thresholds although some limited beneficial effect was detected for some animals after 8 weeks of treatment.

Measurements were made of the ability of nonsimultaneous, contralaterally presented broadband noise maskers to suppress ipsilaterally-evoked compound action potentials (CAPs) of the auditory nerve to 8-kHz tone pips in guinea pigs. Contralateral acoustic stimuli have been shown to suppress responses to ipsilateral stimuli through the action of the medial olivocochlear efferent bundle. In this study, maximum suppression across subjects ranged from 2-6 dB. The ability of the contralateral noise to suppress ipsilateral CAPs was rapidly reduced beginning within minutes of a single intramuscular injection of the aminoglycoside antibiotic gentamicin (150 mg/kg), and, in general, was completely abolished by 1.5-2 h. Changes in suppression were presumably produced by gentamicin-induced blockade of presynaptic calcium channels on the medial olivocochlear efferents and closely follow the pharmacokinetics of gentamicin in perilymph. Reductions in efferent-mediated suppression were observed in the absence of any measurable change in either CAP thresholds to different frequencies or CAP input/output. Suppression was partially restored in some animals after 24 h, and was fully recovered in all subjects within 96 h postinjection. The present studies suggest that measures of efferent function might form the rational basis for an objective means to assess aminoglycoside insult on the cochlea in the absence of hair cell damage. These results also indicate that single injections of gentamicin offer a reversible, non-invasive model in which medial efferent function can be studied in behavioral animals.

A single injection of gentamicin in the awake guinea-pig induces a transient blockade of the suppression by contralateral white noise of tone-pip evoked compound action potential (Smith et al., 1994). We report here the reversible blockade, in exactly the same experimental conditions, of the suppression of click-evoked otoacoustic emissions. These observations confirm the transient action of gentamicin at the medial efferent/outer hair cell synapse, likely via the reversible blockade of calcium channels observed in vitro (Dulon et al., 1989).

In normal adult guinea pigs, evoked potentials recorded at the ipsilateral auditory cortex to monaural high-frequency acoustic stimuli present higher thresholds and lower amplitudes than at the contralateral cortex; in the inferior colliculus, such ipsi-contralateral differences (ICDs) are smaller than in the auditory cortex. Changes in the ICDs were studied after opposite ear injury. Following quasi-complete hair cell destruction induced by sisomicin injection into the contralateral inner ear, threshold ICDs almost disappeared after about two to six days and ipsilateral amplitudes progressively increased in two to three weeks. The occurrence of ICDs at higher auditory centers revealed in this study, indicates peculiar processing of high frequency stimuli in normal guinea pigs. The alteration of ICDs after opposite ear impairment provides a new possibility to study the auditory plasticity in adult animals.

Taurine is recognised as the major osmoregulating amino acid in the brain as well as other organs. Its possible influence on normal cochlear function and on hydropic cochleas of the guinea pig was investigated here. Normally hearing adult guinea pigs were placed on either a taurine-depleting diet (B-alanine) or a taurine-supplemented diet. At the end of 6 weeks the compound action potential audiograms (CAP) in the two groups were similar and appeared not to be different from a third control group receiving a non modified diet which suggested that a taurine-modified diet had not affected normal cochlear function. In hydropic ears the taurine-depleting diet was observed, in some cases, to moderately slow down the early phase of the characteristic progression of the CAP sensitivity loss. These data are in favor of a homeostatic role for taurine in the inner ear of the guinea pig under particular conditions such as osmoregulatory stress.

Confocal laser scanning microscopy and immunofluorescence were used to detect gentamicin at the single cell level in the sensory cells of the guinea-pig hearing organ after a chronic systemic treatment. Gentamicin uptake by the hair cells precedes largely the manifestation of hearing impairment. We demonstrate that for a non-ototoxic treatment the captured gentamicin is eliminated in two phases: one rapid and one slow with approximate half-life of 2 days and 5-6 months, respectively. These novel observations should be of primary importance for new rational approaches to both the mechanism of ototoxicity and therapeutic strategies for the treatment of infectious diseases requiring the aminoglycoside antibiotics.

Recent experiments using autoradiographic and immunohistochemical labelling of gentamicin have demonstrated that GM penetrates specifically into the sensory cells of the inner ear, with a good correlation between the intensity of the labelling and the respective degrees and localisations of ototoxic damages. In the sensory hair cells GM ils localised below the cuticular plate, in an area rich in lysosomes during the ototoxic treatment. This penetration precedes the development of ototoxicity and there seems to be a threshold of intracellular concentration of GM for development of intracellular ototoxic processes. Clearance is very slow since GM can still be observed 11 months after the end of a non toxic treatment (60 mg/kg/day for 6 consecutive days). This observation is of clinical interest, in view of the delayed development of ototoxicity often observed clinically, and with respect to other hazards, including new ototoxic treatments, to which the cells can be exposed while loaded with the aminoglycoside molecule.

Immunodetection of gentamicin (GM) was carried out on surface preparations of the whole organ of Corti from cochleas of guinea pigs treated daily with GM at a dose of 60 mg/kg/day and sacrificed at the end of different treatment periods. Cochlear function was determined just before sacrifice, 24 h after the last injection. Threshold elevations, mainly at high frequencies, were noted only after 10-14 days of treatment. However, the presence of GM was observed much earlier, as early as after the second injection, and specifically in the sensory hair cells. GM labelling was essentially observed in the outer hair cells (OHC) and increased from the apex to the base of the cochlea and from the third to the first row of OHC. GM labelling of inner hair cells was less pronounced and was observed only after the 8th day of treatment. These observations demonstrate that GM specifically enters and accumulates in the sensory hair cells and that the uptake precedes the development of functional and cellular damage which may result from a long-term intracellular cytotoxic action of the molecule.

Presence of gentamicin (GM) in cochlear hair cells was detected by immunohistochemistry in guinea pigs (GPs) cochlea 1, 9 and 41 days after a 6-day treatment with GM at 60 mg/kg/day (s.c.). The number of GPs in each group was respectively 7, 12 and 6. Twelve other non-treated GPs served as controls. Cochlear function was measured, just before sacrifice, by VIIIth nerve compound action potential (CAP) audiograms. Functional and immunohistological evaluations were performed by two independent naïve observers respectively. Functional changes were minimal: only one out of the 25 treated GPs, from the 41-day group, showed significant threshold elevations on high frequencies. Meanwhile GM labelling was observed in most outer hair cells (OHCs) from the three rows of all the treated GPs, with radial and longitudinal gradients, and found similar in the 3 groups. These results 1) confirm that GM is significantly present in OHCs before the development of ototoxicity and 2) indicate that GM accumulates and is maintained inside the OHCs for very long periods of time, i.e. that its clearance from the hair cells, if any, would be very slow.

Alloplastic materials have been used in middle ear reconstruction since 1952. Biologically, biomaterials and specifically ceramics can be defined as being bioinert, bioactive or biodegradable; these terms being descriptive of the biochemical response provoked by the material in the host tissue. Various ceramics, carbons and polymers and their reactions have been studied by the author and his colleagues over the past 5 years utilising animal implantation and fibroblast culture studies aligned with experience from clinical usage. The qualities of the ideal alloplastic implant namely: biocompatibility, stabilisation and incorporation, the consistency of the implant and bioactive bonding and the biofunctionality or mechanics, are applied to the materials currently available. No single alloplastic material fulfills all criteria, the problems encountered in relation to the biomaterials are reported. A better understanding of the microstresses, fatigue and microfractures is necessary, before the ideal alloplastic biomaterial becomes available.

Guinea pigs (GPs) receiving one intra-muscular injection of gentamicin (GM) (150 mg/kg) in which 2 mg of tritiated GM (2 mCi) were incorporated, followed 1.5 h later by an intra-cardiac injection of ethacrynic acid (EA) (30 mg/kg) were sacrificed 25 min, 1, 4 and 24 h after the EA injection. Other GPs were treated with one injection of GM or EA alone and sacrificed 24 h later. Cochlear function was monitored by recording VIIIth nerve compound action potential (CAP) responses to clicks at 70 dB peak-equivalent Sound Pressure Level (pe SPL) and CAP audiograms. At 24 h thresholds were significantly elevated for high frequencies only in GPs treated with the GM/EA combination. GM was revealed in the cochlea and kidney by autoradiography using light and electron microscopy. In the kidney GM was already detected in the proximal tubule cells at 25 min and at 24 h. In the cochlea GM was systematically not observed at 25 min. At 1 h a weak labelling was detected in vessels of the stria vascularis and in sensory cells at the base of the cochlea. At 4 h the labelling disappeared in stria vascularis but increased in the hair cells. At 24 h GM labelling was found exclusively in hair cells, particularly outer hair cells, with a gradient from base to apex and from first to 3rd row, this distribution pattern correlating well with the pattern of threshold changes prominent at high frequencies.(ABSTRACT TRUNCATED AT 250 WORDS)

Guinea pigs were treated with daily single subcutaneous injections of 60 mg gentamicin per kg for 3 weeks. Renal, cochlear and vestibular functions were monitored before, during and after treatment. The degree and onset of gentamicin oto- and nephrotoxocity differed during the treatment period. Alterations to the kidney functions were observed from the first week while the onset of ototoxicity occurred later, at the third and fourth week for the cochlear and vestibular functions respectively. Moreover, when treatment ended, renal function demonstrated signs of recovery, while auditory and vestibular function continued to worsen. Deficits in cochlear function and structural changes (missing outer hair cells) correlated with gentamicin serum concentrations, while vestibular alterations (loss in nystagmic reactions) did not. No distinct relationship could be established between auditory and vestibular loss and the renal parameters monitored. The results suggest that gentamicin-induced nephro- and oto-toxicity are dissociated phenomena and that cochleotoxicity was dependent on aminoglycoside serum level.

An experimental study of the electrical stimulation of the guinea pig cochlea is made using an electrode on the round window for both stimulation and recording. The neural response is separated from the electrical artifact with a masking procedure combined with a low amplification, "statistical" averaging method [Charlet de Sauvage et al., Hear. Res. 2, 343-346 (1980)]. The high electrical impedance required for recording physiological responses implies the use of a current pulse generator. Monitoring of evoked potentials from the auditory cortex provides evidence that the effects of electrical stimulation (and of masking noise) are of auditory origin. The electrically evoked round window response is of very short latency (less than 0.2 ms). There is a response threshold for both electrical stimulus and masking noise. The response amplitude varies monotonically as a function of masking noise or electrical stimulus intensity. Experiments with high-pass noise masking suggest that the electrical stimulus is mainly acting on basal fibers. The response latency and waveform are independent of electrical stimulus intensity, repetition rate, masker level, or spectrum. Little intersubject variation is noted. Our experiments (reciprocal forward masking by electrical and acoustic stimuli) suggest that a direct, instantaneous electrical stimulation of the fibers occurs. We believe that this response to electrical stimulation represents the mean unit response of the auditory nerve fibers. This approach may be useful in the separate study of cochlear and VIIIth nerve functions and in the analysis (deconvolution) of the acoustically evoked compound AP.

Measures of cochlear selectivity can be obtained from compound responses using tone-on-tone masking procedures [Dallos and Cheatham, J. Acoust. Soc. Am. 59, 591--597 (1976)]. For the normal guinea pig, cochlear fiber tuning is sharper by a factor of 1.8 than AP tuning curves using simultaneous masking (threshold criterion = 25% N1 amplitude reduction). Anesthesia does not appear to affect AP tuning. In pathological cochleas, AP tuning is broadened by a factor of 2--3, and differences between forward and simultaneous masking curves are reduced. Tuning changes can sometimes occur without threshold elevation. AP tuning curves were obtained from humans during transtympanic electrocochleography. For subjects with near normal thresholds, Q10dB values (simultaneous masking) are approximately 2.3 at 2 kHz, 3.6 at 4 kHz, and 4.7 at 8 kHz. Using the relationship between cochlear fiber tuning and AP tuning in the guinea pig, estimates of human cochlear fiber tuning are 4.2 at 2 kHz, 6.5 at 4 kHz, and 8.5 at 8 kHz. Patients with threshold elevations of more than 30 dB resulting from cochlear deafness have AP tuning curves less sharply tuned by a factor of 2--3.

Regional selectivity of gentamicin (GM) ototoxicity was studied in guinea pigs (GPs) using electrophysiological, morphological, autoradiographic and immunohistological observations following combined treatment with GM (150 mg/kg i.m.) and ethacrynic acid (EA) (30 mg/kg i.c. or i.v., 1.5 h after GM injection). The GPs were either continuously stimulated every 5 min with a series of 256 clicks (70 dB peSPL, 10/s) during 3 h for monitoring fast changes in VIII nerve compound action potential (CAP) after the EA injection, and thereafter kept in the animal quarters (background noise of 60 dB SPL) (group I), or similarly monitored for only 10 min after the EA injection and thereafter kept in a soundproof room (around 0 dB SPL) (group II). Whenever GM labelling was observed it was localized only in the sensory hair cells. From 3 h after EA injection, the GPs in group I presented threshold elevations in the high-frequency region, which progressed to 60-80 dB at all frequencies at and after 48 h. Parallel to the threshold pattern, GM uptake in outer hair cells (OHCs) was seen with an increasing concentration from apex toward base from 3 to 24 h, while after 48 h almost all OHCs were destroyed and inner hair cells (IHCs) were marked by GM. In group II no changes in CAP thresholds were observed until more than 24 h, although GM was detected in the hair cells from 6 h on. At this early stage, the distribution of GM lacked a clear pattern, particularly without a clear apex-base gradient, and GM deposits were found only around the basal body. However in both groups, in late stage (greater than 24 h), the base-apex gradient was more pronounced and GM was found throughout the cell body, with a marked concentration below the cuticular plate. These results suggest that GM may penetrate hair cells around the basal body and that activating the cells by sound potentiates both GM uptake and its intracellular toxicity.

Long-term treatment of guinea pigs with the diuretics chlorthalidone and acetazolamide, following the experimental obstruction of the endolymphatic duct, was assessed using chronically implanted round window cochlear electrodes. The diuretic chlorthalidone appeared to curb the progressive low-frequency sensitivity loss during the first 4 weeks following surgery, as compared with animals receiving the diuretic acetazolamide or no treatment. However, this apparent beneficial effect decreased after 4 weeks and was not apparent at 14 weeks post-induction. On the other hand, morphological control at the end of 14 weeks confirmed a marked reduction in hydrops in the chlorthalidone-treated animals. The data clearly demonstrate a dissociation between hydrops and the development of hearing loss and suggest that the augmenting endolymph volume is only one of several contributing factors to the deteriorating auditory function in experimental hydrops.

Acoustically evoked neural activity has been recorded from the brainstem and auditory cortex of guinea pigs after complete destruction of the organ of Corti by the aminoglycosidic antibiotic amikacin. These responses to sound differ in important respects from the evoked potentials normally recorded from the auditory pathways. At the brainstem level they resemble the potentials reported by others after stimulation of the vestibular nerve.

This paper describes a method of investigation of the peripheral vestibular system. Electrical stimulations (ES) are applied on the round window of chronically implanted guinea pigs, with and without vestibular stimulation (either per or post rotational accelerations). The whole nerve action potential recorded at the output of the internal auditory meatus, the difference between the two conditions, reveals the change in electrical excitability and thus presumably in discharge rate of vestibular fibres determined by rotational stimuli. This electrical vestibular action potential (EVAP) presents as a mainly monophasic potential with a peak latency to ES of about 0.3 ms. Right and left accelerations versus rest are shown to give responses with opposite polarity, reflecting the inhibitory and excitatory influences of these opposite accelerations, whereas the transfer function of EVAP versus acceleration amplitude appears roughly linear. These observations appear altogether coherent with published mechano-physiological data about individual discharge rates of canal fibres. Also relative amplitudes of acoustical and vestibular responses are in agreement with related number of fibres in the two systems. This EVAP could be the basis of a quantitative evaluation method for the vestibular system, the electrovestibulogram (EVG).

We have previously shown by chronic implantation of round window electrodes, that after the experimental induction of endolymphatic hydrops by surgical blockade of the endolymphatic canal, in the guinea pig, there is a low/mid-frequency fluctuant hearing loss, followed by a very high frequency loss and after a few months the audiogram is flat. This evolution reproduces exactly that observed in Meniere's patients. The aim of this study was to investigate the evolution of one aspect of vestibular function in parallel with audiogram changes. The nystagmic responses in the dark were tested every month during the 4 months which followed hydrops induction. There was considerable variation in the number of evoked saccades for different control animals and even between different recordings for the same control animal. However, in general, the number of saccades to right and left rotation was symmetrical for control animals, whereas for hydropic animals there was a period, within the first 2 months post-operation, when there was a reduction of saccades to the left (operated side) with sometimes an increase to the right. This asymmetry which occurred in the period of fluctuant hearing losses did not however appear synchronized with audiogram fluctuations. The nystagmic responses tended to become symmetrical over the 4 months post-operation. Oral administration of glycerol to animals whose nystagmus was asymmetric brought the vestibular response towards symmetry in 50% of the cases. Although episodes of vertigo, as known in Meniere's disease, were never observed in operated guinea pigs, these data indicate that they have some functional vestibular disturbance.

The compound action potential (CAP) recorded at the round window has for some time been investigated in order to determine the underlying temporal spike sequence: the firing probability density (FPD) function of the fibres. Charlet de Sauvage et al. [(1980) Hear. Res. 2, 343-346; (1983) J. Acoust. Soc. Am. 73, 616-627] have previously described a close estimate of the far-field unit response (UR) by presenting to the ear a transient electrical stimulation combined with an acoustical masking noise. A linear model of UR, free of noise or extrinsic potentials and based on both experimental data and on the dipole principle of Teas et al. [(1962) J. Acoust. Soc. Am. 34, 1438-1459], was proposed [(1985) Hear. Res. 18, 121-125]. Using the UR model, deconvolutions were performed on CAPs recorded in various conditions of stimulation and of cochlear pathologies in the guinea pig. The principal findings were: (1) a plateau was observed in the amplitude function around 60-70 dB above normal threshold both for click-evoked CAPs and for the corresponding FPDs; (2) FPDs resulting from the deconvolution of high-frequency derived responses showed 2 main peaks with a 0.7 ms interval followed by a second homologous pair with lower amplitude. Convolution computations confirmed the need for the second set with peaks 2 and 3 being only 0.3 ms apart. These findings are interpreted in terms of mechanical excitation pattern.

Anatomical and physiological studies of brainstem acoustic nuclei involving the classical ascending auditory pathway or the cerebellar and reticular pathways imply that all afferents from the cochlea terminate in the cochlear nucleus. In the experimental pathology of complete and selective destruction of the cochlea in the guinea pig acoustic responses still evoked at mid and high intensities, demonstrated to come from the saccule, show a pattern of far field evoked brainstem potentials quite different from that of normal animals. Intracranial electrophysiological investigations of the brainstem were undertaken in such pathological animals and in normal guinea pigs for comparison. In both cases acoustic responses were recorded at the base of the vestibular nucleus, showing a first peak corresponding to an eighth nerve projection and after a synaptic delay a second peak of local activation. In normal animals acoustic responses from the vestibular nucleus showing normal threshold and tuning curves may represent a direct projection from the cochlea.

After complete destruction of cochlear but preservation of vestibular hair cells in the guinea pig acoustically evoked responses can still be recorded from the round window up to the auditory cortex. At all levels these responses differ from those observed in normal animals but their frequency sensitivity and selectivity make them akin to responses from auditory organs. In a series of experiments a complete cochlear destruction was combined with a total or partial destruction of the vestibule. After complete cochlear and vestibular hair cell destruction no acoustic response could be recorded. But in cases of total cochlear and drastic ampullar and utricular destruction together with an almost undamaged saccular sensory epithelium the same peculiar acoustic responses could be observed. These results support the hypothesis of a functional acoustic reception by the saccule in a mammal.

VIII nerve compound action potentials evoked by electrical stimulation are decomposed in series of pure-tone derived potentials using masking. These potentials are in turn decomposed in a series of contributions from various frequency bands by adding high-pass masking noises. The amplitudes of the derived signals are thought to be proportional to the number of fibres activated by the masking acoustical stimulations. Thus the density of nerve fibres stimulated along the basilar membrane by pure tones as a function of frequency and intensity can be estimated.

Averaged VIIIth nerve and brainstem potentials are recorded in normal guinea pigs in response to electrical stimulation of the ear presented once without and once with a simultaneous masking continuous pure tone. The waveform difference yields a well synchronized 'derived signal' which is the response to the electrical stimulation of only the group of fibers masked by the pure tone. Study of very low frequency activation is a new possibility brought by this method which is efficient whatever the sound frequency. Pure tone thresholds obtained are similar to previous measurements. Response amplitudes are likely a measure of the number of fibers stimulated by the sounds.

In guinea pigs that had been treated with very large doses of the aminoglycoside amikacin (14 x 450 mg/kg/day, i.m.) clear, short-latency responses to various click stimuli could be recorded at the round window. When the same cochleas were examined histologically, no outer or inner hair cells could be found along the entire length of the basilar membrane, save for a very few outer hair cells remaining at the apex. The response patterns resembled that of the compound action potential, and various characteristics suggest that they were of neural origin. Vestibular function, investigated by electronystagmography during rotation, appeared normal, as did most of the saccular and utricular hair cells. Transmission electron microscopy revealed a significant number of cochlear nerve fibres still innervating the remnants of Corti's organ. In other cochleas with similarly extensive destruction induced by another aminoglycoside (sisomycin, 14 x 125 mg/kg/day, 15 days as well as 3 months post-Rx), no responses could be recorded from the round window. Cochleas that were less affected, with the upper turns preserved, gave only small, long-latency responses. These preliminary observations are confirmed by further experiments now in progress. They suggest that unless a considerable number of inner hair cells remained undetected in the lower basal turn, a possibility that appears highly unlikely, there was either a direct mechanical excitation of cochlear nerve fibres, or an acoustical stimulation of vestibular sense organs.

Experiments were performed on guinea pigs to assess the eventual ototoxic side effects of a new glycopeptide antibiotic called teicoplanin. For a better validation, in the same study other animals were treated with tobramycin, a well known ototoxic antibiotic of the aminoglycoside family, and a group of control animals was also constituted. Antibiotic treated animals were injected with the high dose of 90 mg/kg/day during 21 consecutive days. Electrophysiological measures of equilibrium and acoustic functions were taken and morphological evaluations of the cochleas were performed. The results indicate that in these animal experiments teicomycin appears to be clearly toxic for the vestibule and the cochlea.