The quinoxalinediones DNOX, CNOX and two related congeners suppress hair cell-to-auditory nerve transmission

We tested 6,7-dinitroquinoxaline-2,3-dione (DNQX); 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX); 6,7-dichloro-3-hydroxy-2-quinoxalinecarboxylic acid (DHQC); and 3-hydroxy-2-quinoxalinecarboxylic acid (3HQC), new kainate and quisqualate receptor antagonists, upon cochlear potentials in guinea pig. Perilymph spaces of guinea pig cochleae were perfused with artificial perilymph solutions containing up to 1000 microM concentrations of DHQC and 3HQC, and 500 microM concentrations of DNQX and CNQX, at a rate of 2.5 microliters/min for 10 min. Cochlear potentials evoked by 10 kHz tone bursts of varying intensity were recorded from the basal turn scala vestibuli. Cochlear perfusion of the four drugs resulted in a dose-related suppression of the compound action potential of the auditory nerve (CAP; N1-P1), a prolongation of N1 latency at suprathreshold levels, an elevated CAP threshold, and a decreased N1 latency at CAP threshold. None of the drugs had significant effects on cochlear microphonics (CM) or the summating potential (SP). EC50 values (concentrations causing a 50% reduction in CAP amplitude at 68 dB SPL) were 8 microM for DNQX, 30 microM for DHQC, 35 microM for CNQX, and 1 mM for 3HQC. Results support the hypothesis that kainate and quisqualate receptors are involved in neurotransmission between the hair cell and afferent nerve.     

Effects of nimodipine, an L-type calcium channel antagonist, on the chicken's cochlear potentials

At most synapses in the brain, neurotransmitter release depends on N-type or P/Q-type calcium channels. However, available in vitro experimental data suggest that there exist almost exclusively L-type calcium channels in sensory hair cells of most species. To test whether chicken hair cells depend on L-type calcium channels for neurotransmitter release, we examined the effects of nimodipine, a selective L-type calcium channel antagonist, on acoustically evoked cochlear potentials in 10-15 week old chickens in vivo. Diffusion of nimodipine into scala tympani significantly elevated threshold, dramatically decreased the amplitude and increased the latency of the compound action potential within 20 min of drug application. The summating potential was also significantly reduced in amplitude, but the cochlear microphonic was relatively less affected. All the effects were reversible after nimodipine was washed out with artificial perilymph except that the cochlear microphonic amplitude remained decreased. Application of omega-conotoxin GVIA, an N-type calcium channel antagonist and agatoxin Tk, a P-type calcium channel antagonist had no observable effects on the cochlear potentials. These results suggest that L-type calcium channels control neurotransmitter release from avian hair cells.     

An interaction between PPADS, an ATP antagonist, and a moderately intense sound in the cochlea

In the organ of Corti ionotropic receptors for ATP (ATPRs) on cells that are bathed by perilymph have been suggested to modulate cochlear mechanics. The purpose of the present study was to test the hypothesis that endogenous extracellular ATP acting through ATPRs is involved in modulating cochlear mechanics during moderately intense sound exposure. Guinea pigs were exposed to either: (1) a perilymphatic administration of pyridoxal-phosphate-6-azophenyl-2', 4'-disulfonic acid (PPADS, 1 mM), an ATP antagonist; (2) a moderately intense sound (6.7 kHz tone, 95 dB SPL, 15 min); or (3) a combination of both the PPADS and the sound. The effects on cochlear potentials (cochlear microphonic, CM; negative summating potential, SP; compound action potential of the auditory nerve, CAP; and N(1) latency) evoked by a 10 kHz tone pip were monitored. PPADS alone reduced the CAP and the SP and increased N(1) latency. The intense sound alone reduced the CAP and SP. The combination of PPADS with the intense tone induced reversible effects on cochlear potentials that were greater than induced by either treatment alone. The effect on N(1) latency and low intensity CM was a potentiation since the effect was greater than a simple addition of the effect of either treatment alone. The effects of the combination treatment on CAP, SP and high intensity CM were not different from additive. Results are consistent with the hypothesis that ATPRs in the organ of Corti are involved in modulating cochlear mechanics during moderately intense sound exposure.     

Caffeine and ryanodine demonstrate a role for the ryanodine receptor in the organ of Corti

The hypothesis that the release of Ca(2+) from ryanodine receptor activated Ca(2+) stores in vivo can affect the function of the cochlea was tested by examining the effects of caffeine (1-10 mM) and ryanodine (1-333 microM), two drugs that release Ca(2+) from these intracellular stores. The drugs were infused into the perilymph compartment of the guinea pig cochlea while sound (10 kHz) evoked cochlear potentials and distortion product otoacoustic emissions (DPOAEs; 2f(1)-f(2)=8 kHz, f(2)=12 kHz) were monitored. Caffeine significantly suppressed the compound action potential of the auditory nerve (CAP) at low intensity (56 dB SPL; 3.3 and 10 mM) and high intensity (92 dB SPL; 10 mM), increased N1 latency at high and low intensity (3 and 10 mM) and suppressed low intensity summating potential (SP; 10 mM) without an effect on high intensity SP. Ryanodine significantly suppressed the CAP at low intensity (100 and 333 microM) and at high intensity (333 microM), increased N1 latency at low intensity (33, 100 and 333 microM) and at high intensity (333 microM) and suppressed low intensity SP (100 and 333 microM) and increased high intensity SP (333 microM). The cochlear microphonic (CM) evoked by 10 kHz tone bursts was not affected by caffeine at high or low intensity, and ryanodine had no effect on it at low intensity but decreased it at high intensity (10, 33, 100 and 333 microM). In contrast, caffeine (10 mM) and ryanodine (33 and 100 microM) significantly increased CM evoked by l kHz tone bursts and recorded from the round window. Caffeine (10 mM) and ryanodine (100 microM) reversibly suppressed the cubic DPOAEs evoked by low intensity primaries. Overall, low intensity evoked responses were more sensitive and were suppressed to a greater extent by both drugs. This is consistent with the hypothesis that release of Ca(2+) from ryanodine receptor Ca(2+) stores, possibly in outer hair cells and supporting cells, affects the function of the cochlear amplifier.     

Effects of a calcium channel blocker on spontaneous neural noise and gross action potential waveforms in the guinea pig cochlea

The effects of the L-type Ca2+ channel blocker nimodipine on the spectrum of the spontaneous neural noise (SNN) and the waveform of the gross sound-evoked compound action potential (CAP) were investigated by perilymphatic perfusion in the guinea pig cochlea. Both the SNN and the CAP were reversibly suppressed by nimodipine. The percentage reduction in SNN was dose-dependent in a manner very similar to the results obtained with the measures of CAP threshold changes. The reduction in the peak SNN caused by 10 microM nimodipine was the same as that caused by 500 microM kainic acid, which totally eliminated any neural responses. For 1 microM nimodipine there was an apparent dissociation between the SNN and CAP changes such that the SNN could be markedly suppressed with only very small changes in CAP thresholds. These results imply that spontaneous release of neurotransmitter from the inner hair cell is more sensitive to block of calcium channels than evoked release. There was no evidence for any marked shift caused by nimodipine, in the position of the main (900 Hz) spectral peak in the SNN. Comparison of the CAP waveform before and after nimodipine perfusion showed that the CAP waveforms were unchanged despite the change in sensitivity. These data do not support the notion of any significant postsynaptic site of action of nimodipine. The data hence provide further support for an exclusively presynaptic role for L-type Ca2+ channels in the regulation of both evoked and spontaneous neurotransmitter release from inner hair cells.     

水杨酸钠致耳蜗电生理改变

目的：了解水杨酸钠对耳蜗功能的影响，探讨其可能的作用机制。方法：采用水杨酸钠在活体豚鼠耳蜗进行灌注，同时监测豚鼠耳蜗电位中的复合动作电位(CAP)和总合电位(SP)的阈值及输入输出曲线变化，对耳蜗功能进行评估。结果：水杨酸钠可引起CAP和SP阈值同步改变，两者的变化呈一一对应关系。结论：水杨酸钠主要作用于耳蜗外毛细胞，通过影响外毛细胞的能动作用而影响耳蜗功能。     

2-Amino-4-phosphonobutyric acid receptors are not involved in synaptic transmission from hair cells to auditory neurons

The chemical 2-amino-4-phosphonobutyric acid (APB, AP4), an excitatory amino acid antagonist, was perfused through the guinea-pig cochlea while monitoring various cochlear potentials. The drug (0.6-10 mM) had no effect on the magnitude of the compound action potential of the cochlear nerve, N1 latency, cochlear microphonics, or the summating potential (SP). The results are consistent with the hypothesis that the APB receptor is not involved in neurotransmission between cochlear hair cells and afferent nerve fibers.     

Cytochalasin D suppresses sound evoked potentials in the guinea pig cochlea

Cytochalasin D (CD), an inhibitor of actin polymerization, was perfused through the guinea pig cochlea while monitoring various cochlear potentials. CD (10(-7)-10(-5) M) reduced the magnitude of the compound action potential of the cochlear nerve and the summating potential, and increased N1 latency. The cochlear microphonic potential was the least sensitive potential with only a slight effect being observed at 10(-5) M. The results are consistent with the hypothesis that actin has a role in cochlear function.     

The effect of adenosine on cochlear potentials in the guinea pig

The effect of adenosine on cochlear potentials was examined in the guinea pig. Perilymphatic perfusion with 10^–4 M adenosine produced a significant decrease in the amplitudes of cochlear microphonics, negative summating potential (-SP) and compound action potential (CAP) and significant prolongation of N1 latency with no change in the endocochlear potential. The decreases in the amplitudes of -SP and CAP caused by adenosine were dose-dependent. Perilymphatic perfusion with an inactive analogue, 8-bromoadenosine, produced no changes in the cochlear potentials. The A1-receptor agonist, 2-chloroadenosine, produced a similar change in cochlear potentials to adenosine, while no changes were produced by the A2-receptor agonist ,5-(N-ethylcarboxamido)-adenosine. These results suggest that adenosine may have a modulatory function through an Al receptor in the cochlea.     

Phosphorothioate oligodeoxynucleotides can selectively alter neuronal activity in the cochlea.

A growing body of evidence indicates that extracellular adenosine triphosphate (ATP) may have a major role in cochlear function. Antagonists of ionotropic ATP receptors (P2X2) have significant effects on cochlear potentials and distortion product otoacoustic emissions (DPOAEs). We tested whether antisense oligodeoxynucleotides (ODNs) would mimic the functional deficiencies induced by the ATP antagonists through binding to P2X2 ATP receptor mRNA and thereby reduce the number of ATP receptors expressed in the membrane of the cells. Both a phosphorothioate ODN (S-ODN) antisense and a phosphodiester ODN (P-ODN) antisense to the P2X2 sequence and random sense ODNs containing 21 nucleotides were administered chronically (7 days) to the guinea pig cochlea via the perilymph compartment. Sound evoked cochlear potentials (cochlear microphonic; summating potential; compound action potential of the auditory nerve, CAP; latency of the first negative peak in the CAP, N1 latency) and DPOAEs were monitored to assess the effects of the ODNs. Results indicate that the phosphorothioate derivatives of both the antisense and random sense ODNs suppressed the CAP and prolonged the N1 latency with no significant effect on the other parameters. The P-ODNs had no effect. Since both the antisense and random sense S-ODNs had the same effect, we conclude that the S-ODNs affected neuronal function in a manner that did not involve binding to the ATP receptor mRNA.     

Thapsigargin suppresses cochlear potentials and DPOAEs and is toxic to hair cells

Thapsigargin, a drug that inhibits sarco-endoplasmic reticulum Ca(2+) ATPases (SERCAs), was infused into the perilymph compartment of the guinea pig cochlea in increasing concentrations (0.1-10 microM) while sound evoked cochlear potentials were monitored. Thapsigargin significantly suppressed the compound action potential of the auditory nerve, cochlear microphonics, and increased N(1) latency at low (56 dB SPL) and high intensity (92 dB SPL) levels of sound, suppressed low intensity sound evoked summating potential (SP) and greatly increased the magnitude of the high intensity sound evoked SP. At 10 microM, the drug suppressed the cubic distortion product otoacoustic emissions (2f(1)-f(2)=8 kHz, f(2)=12 kHz) evoked by both high and low intensity primaries (45, 60, 70 dB SPL). Thapsigargin (10 microM; 30 min) increased the endocochlear potential slightly (5 mV). In chronic animals, thapsigargin (10 microM; 60 min) destroyed many outer hair cells and some inner hair cells, especially in the basal turns. These effects are consistent with the hypothesis that the inhibition of the SERCAs affects the function of the cochlear amplifier and outer hair cells to a greater degree than it affects other functions of the cochlea.     

Effects of a dopaminergic agonist in the guinea pig cochlea

This study investigates the role of dopamine, a putative lateral efferent neurotransmitter/modulator, in cochlear physiology and physiopathology. Cochlear potentials were recorded in guinea pigs after intracochlear perfusion of increasing doses (0.1–1 mM) of piribedil, an agonist of the D2/D3 receptors. A dose-dependent reduction in the amplitude of auditory nerve compound action potential (CAP) was observed, predominantly at high-intensity tone-burst stimulations, and without significant effect on CAP threshold. There was no variation of cochlear microphonic and summating potential.

When 1 mM piribedil was perfused into the cochlea during continuous 130 dB SPL pure tone exposure (6 kHz, 15 min), CAP threshold shifts were significantly less than in control animals with artificial perilymph-perfused cochleas. No dendritic damage was observed, although there was evident hair cell damage. Similarly, radial dendrites were clearly protected against ischemia-induced damage when 1 mM piribedil was applied prior to a 10-min ischemia.

These results suggests that dopamine modulates the activity of radial afferent fibers via D2/D3 receptors. The protective effect of piribedil during acoustic trauman or ischemia suggests that this modulation corresponds to a prevention of excitotoxicity due to dysfunction of inner hair cell neurotransmission.     

The effects of moderate cooling on gross cochlear potentials in the gerbil: basal and apical differences.

Changes in the threshold of the compound action potential (CAP) response in the gerbil to low- and high-frequency tonebursts were monitored during uniform cooling of the cochlea by 7-8 degrees C below normal body temperature. Recordings of the endocochlear potential (EP), cochlear microphonic (CM), and summating potentials (SP) were also obtained from the base and apex of the cochlea under the same conditions. Cooling-related changes in the CAP, as well as the CM and SP response obtained near the best frequency of the recording location, were greater in the base than in the apex. In contrast, reductions in the EP appeared uniform throughout the cochlea. Thus the greater vulnerability of CAP thresholds in the base does not result from a greater vulnerability of the stria vascularis in this region. Our results suggest that the enhanced susceptibility to cooling of the CAP in the cochlear base reflects changes in hair cell mechanisms.     

The action of substance P methyl ester on cochlear potentials in the guinea pig

The action of the substance P agonist, substance P methyl ester (SPME) on cochlear potentials was examined in the guinea pig. Previous studies have shown that SPME is a selective agonist for neurokinin 1(NKI) receptor. Perfusion with SPME at a concentration of more than 10-⁶M produced an increase in the amplitudes of the compound action potential and negative summating potential in a dose-dependent manner. N1 latency showed a tendency to be shortened, but this change was not significant. Amplitudes of the cochlear microphonics and endocochlear potential remained unchanged. Substance P fragment 7–11, an inactive analogue, produced no changes in the cochlear potentials. In contrast, the substance P antagonist [D-Pro²,D-Trp^7,9]-SP blocked the action of SPME on the cochlear potentials. These results suggest that substance P may modulate neurotransmission through NK1 receptors in the cochlea.     

Acoustic input-admittance of the alligator-lizard ear: nonlinear features

The electrically evoked compound action potential (CAP) of the auditory nerve exhibits two peaks, termed N0, at 350 microseconds latency, and N1, at 550 microseconds latency. At low stimulus intensities the CAP consists solely of the long latency N1 peak. As the stimulus strength is increased the higher threshold N0 appears. At high stimulus intensities N1 disappears and only the N0 component of the CAP remains. It is postulated that N1 represents action potentials propagated from the dendritic processes of the auditory neurons and that N0 represents action potentials initiated on the axons of these cells. The N1 peak exhibits anomalous refractory behavior which can be identified in the electrically evoked auditory brainstem response (EABR). That behavior may be useful diagnostically in assessing the extent of dendrite degeneration in cochlear implant candidates and users.     

Application of a neuroprotective ACTH(4–9) analog to affect cisplatin ototoxicity: an electrocochleographic study in guinea pigs

Ototoxicity and neurotoxicity are among the most serious side-effects of cisplatin therapy. Previous experiments have shown that neurotoxicity can be delayed or prevented by treatment with the melanocortin-derived peptide ORG 2766, and ACTH_4–9 analog. A remedy against ototoxicity is not available. In this study we describe cisplatin-induced abnormalities in cochlear potentials in guinea pigs. These included changes in compound action potential (CAP), cochlear microphonics (CM) and summating potential (SP) at frequencies from 500 Hz to 16 kHz. Cisplatin (2 mg/kg for 8 days) reduced CAP amplitude with the effect becoming more pronounced at higher frequencies. Cisplatin also reduced CM and SP. Concurrent treatment with ORG 2766 prevented cisplatin ototoxicity partially or completely in four out of ten animals. In the other six animals the effects were comparable to those seen in control animals not treated with the peptide. The protective effects found with this neurotrophic peptide warrant further experimentation.     

Summating potential in the guinea pig cochlea after perilymphatic perfusion with arginine-vasopressin

We hypothesized that arginine-vasopressin (AVP), the natural vasopressin in the guinea pig, might increase the cochlear summating potential, indicating an increase in endolymphatic volume. Guinea pig cochleas were perfused with artificial perilymph for 15 min, with or without AVP (2 x 10(-6) M). In 1 group of animals, summating potentials (SP), compound action potentials and cochlear microphonics evoked by 2, 4 and 8 kHz tone bursts were measured with an apically placed electrode 15 min, 1 and 2 h after perilymphatic perfusion. In another group of animals the SP and endocochlear potential (EP) were measured simultaneously in the scala media during and after perfusion. In both groups the SP had increased significantly 15 min after perfusion with AVP and this increase was reversible. At the concentration of AVP used the increase in SP was not related to EP alterations. On light microscopic examination of the cochlea no evident increase in scala media volume could be detected. The increase in the SP (a sensitive indicator of acute endolymphatic hydrops) after perfusion with AVP suggests that this neuropeptide plays a role in the regulation of the ion and fluid balance in the cochlea.     

Same location of the source of P1 of BAEPs and N1 of CAP in guinea pig

The purpose of this study was to point out the location of N1 and N2 of the compound action potential (CAP) and the first waves of the brainstem auditory evoked potentials (BAEPs) by the mean of the section of the eighth nerve at the porus acousticus in guinea pigs. Our results agree with most of the studies in the literature. In the absence of lesion of the internal auditory artery, the section of the cochlear nerve induced the persistence, alone, of the P1 of BAEPs in which latency was similar to initial P1 before section, and the subsequent waves disappeared. Simultaneously, a monophasic negative potential (N1) of the auditory nerve remained without N2. When the section included the internal auditory artery all BAEPs and CAP components disappeared simultaneously and very suddenly just after the section. These results definitively excluded a proximal contribution of the cochlear nerve in the N1 of CAP and P1 of BAEPs. The N2 of CAP and P2 of BAEPs are not generated, even in part, in the intracochlear or intrapetrous portion of the cochlear nerve.     

Electrocochleography possibilities in the differential diagnosis of hydrops and neural hearing loss

Enhancement of the click-evoked summating potential causing a pathological summation to compound action potential ratio (SP/CAP) in patients with endolymphatic hydrops has been described by several groups. An increased SP/CAP ratio is found also in some patients with a tumour of the cerebellopontine angle. The study of further parameters of the SP/CAP complex, such as the CAP latency and the width of the complex, permit an electrocochleographic differentiation between both forms of hearing loss. The N2 amplitude showed no diagnostic significance.     

Effect of Endoplasmic Ca2+-ATPase Inhibitors on Cochlear Potentials in the Guinea-pig

The effect of endoplasmic Ca²⁺-ATPase inhibitors on cochlear potentials was examined in the guinea-pig. Perilymphatic perfusion with thapsigargin (10^-6M) produced a significant decrease in the amplitudes of cochlear microphonics, negative summating potential and compound action potential, and a significant prolongation of N₁ latency with no change in the endocochlear potential. These changes were all dose dependent. Another endoplasmic Ca²⁺-ATPase inhibitor, cyclopiazonic acid (10^-5M), produced the same effects as thapsigargin on cochlear potentials. These results suggest that endoplasmic Ca²⁺-ATPase inhibitors may have inhibitory functions on cochlear potentials.