Richard P. Bobbin

Contralateral sound suppresses distortion product otoacoustic emissions through cholinergic mechanisms

Presentation of an acoustic signal to one ear can suppress sound-evoked activity recorded at the opposite ear. The suppression appears to be mediated by medial olivocochlear (MOC) efferent neurons synapsing with outer hair cells (OHCs) and acting through the MOC neurotransmitter, acetylcholine (ACh). The purpose of the present investigation was to study the suppression of distortion product otoacoustic emissions (DPOAEs) by contralateral sound and to examine whether the suppression could be blocked by known antagonists of olivocochlear (OC) efferent activity. Urethane-anesthetized guinea pigs were used. Perilymph spaces of ipsilateral cochleae were alternately perfused with artificial perilymph and drugs at 2.5 microliters/min for 10 min. After each period of perfusion, DPOAEs were measured before, during and after contralateral wideband noise (WBN) stimulation. Pre-perfusion, contralateral WBN attenuated the ipsilateral DPOAEs between 1-3 dB. This suppression was blocked reversibly by strychnine (10 microM), curare (10 microM) and atropine (20 microM), known antagonists of OC efferent activity. These results confirm the findings of Puel and Rebillard (1990) that contralateral WBN can suppress DPOAEs in anesthetized guinea pigs. Furthermore, results suggest that this efferent control of the cochlear mechanical response can either be mediated by both nicotinic and muscarinic cholinergic receptors, or that a single receptor with as yet undescribed structure and pharmacology mediates effects seen.

Intracochlear salicylate reduces low-intensity acoustic and cochlear microphonic distortion products ☆

Salicylate is well-known to produce reversible hearing loss and tinnitus. The site and mechanism of salicylates ototoxic actions, however, remain unresolved. Recent experiments demonstrating primarily low-intensity effects on cochlear afferent outflow and effects on otoacoustic emissions (OAEs) suggest that salicylate acts to compromise active, energy-enhancing processes within the cochlea (i.e., the active process). We tested this hypothesis by examining the effect of salicylate on distortion product emissions. Distortion product responses to two-tone stimulation were monitored in the guinea pig before, during, and after intracochlear administration of increasing concentrations of salicylate (0.6-5 mM). These responses were recorded as acoustic signals in the ear canal spectrum (ADP), and as present in the cochlear microphonic (CM) recorded from a wire in basal turn scala vestibuli (CMDP). We also recorded the CM response to a single tone. Cochlear perfusion of salicylate resulted in a dose-responsive reduction in ADPs that was greater for low intensities of stimulation. CMDPs also demonstrated a concentration-dependent reduction at low intensities, but were increased slightly, though not significantly, by salicylate when elicited by high intensity primaries. CM was essentially unchanged by intracochlear salicylate. These results are consistent with an action of salicylate that involves the outer hair cells (OHCs) and are in harmony with the hypothesis that salicylate may selectively compromise the active process.

Comparative actions of quisqualate and N-methyl-d-aspartate, excitation amino acid agonists, on guinea-pig cochlear potentials

We examined dose-dependent changes in the amplitude of guinea-pig cochlear microphonic potentials (CM), summating potentials (SP) and compound auditory nerve action potentials (CAP) produced after perfusing perilymphatic scalae with artificial perilymph containing either the transmitter candidate, L-glutamate; one of the excitatory amino acid agonists, quisqualate, kainate, N-methyl-D-aspartate (NMDA) or D-glutamate; or the control, alpha-ketoglutarate. None of these compounds significantly altered CM or SP. Kainate abolished CAP, but only partial suppression occurred using maximal effective doses of quisqualate (67%) or L-glutamate (82%). The remaining compounds had only marginal effects on CAP. The potency of quisqualate (EC50 = 14.8 microM) exceeded that of both kainate (EC50 = 66.9 microM) and L-glutamate (EC50 = 1.41 mM). These data suggest the presence of neuronal, possibly postsynaptic, excitatory amino acid receptor subpopulations which are preferentially sensitive to quisqualate and to kainate, but not to NMDA. These findings are discussed in the framework of our hypothesis that the proposed quisqualate and kainate receptors are normally activated by an endogenous excitatory amino acid such as L-glutamate which the hair cells release as a neurotransmitter.

Kynurenic acid and γ-D-glutamylaminomethylsulfonic acid suppress the compound action potential of the auditory nerve

Kynurenic acid and gamma-D-glutamylaminomethylsulfonic acid, two excitatory amino acid antagonists, were perfused through the guinea pig cochlea while monitoring various cochlear potentials. Both drugs (0.6-10 mM) reduced the magnitude of the compound action potential (CAP) of the cochlear nerve without much effect on other potentials. The results are consistent with the hypothesis that the hair cell transmitter is an excitatory amino acid, possibly L-glutamate.

Intracochlear application of acetylcholine alters sound-induced mechanical events within the cochlear partition

Activation of olivocochlear (OC) efferent fibers has been suggested to alter micromechanical events occurring within the cochlear partition, possibly through an effect of the efferent neurotransmitter (acetylcholine; ACh) on outer hair cells (OHCs). Based on the widely-accepted assumption that otoacoustic emissions reflect OHC activity, we investigated the in vivo influence of ACh on OHCs by studying alterations in emission amplitude with local ACh application. Distortion product otoacoustic emissions (DPOAEs) were measured in anesthetized guinea pigs before, during, and after intracochlear application of ACh (250 microM) with the cholinesterase inhibitor, eserine (20 microM). Perfusion of ACh/eserine was associated with a desensitizing reduction in DPOAE amplitude of approximately 4.4 dB. This reduction was intensity-dependent, with greater and more consistent reductions observed for DPOAEs elicited by low- than by moderate-intensity primaries. The response reduction was not seen during consecutive ACh perfusions performed without an intervening artificial perilymph wash, and was effectively blocked in the presence of pharmacologic antagonists of OC efferent activity (curare, 50 microM; strychnine, 50 microM). Finally, a similar alteration in DPOAE amplitude was never seen during perfusion of the control (artificial perilymph) solution alone. It is argued that these results support the hypothesis that OC efferent activation can alter sound-induced cochlear mechanical events.

Potassium induced release of GABA and other substances from the guinea pig cochlea

Gamma-aminobutyric acid (GABA) has been proposed as a neurotransmitter of a subset of efferent nerve fibers in the mammalian cochlea. We tested this hypothesis by examining if GABA was released by high concentrations of K+ from the guinea pig cochlea. Artificial perilymph solutions containing either normal K+ (5 mM) or high K+ (50 mM) were perfused through the perilymphatic compartment of the guinea pig cochlea while collecting the effluent. Nineteen primary amines including GABA were quantified in the effluent by HPLC. This was carried out in normal animals and in animals pretreated with ethacrynic acid and kanamycin to destroy the organ of Corti. Significantly greater levels of GABA, taurine, glutamate, aspartate, glycine and three unidentified substances appeared in effluent collected during exposure of the cochlea to solutions containing higher K+ than normal K+. Compared to normal animals, destruction of the organ of Corti significantly decreased the K(+)-induced release of GABA, taurine, glutamate, aspartate, glycine and one of the unidentified substances; although significant release of glutamate and taurine still occurred in the destroyed ears. The release of GABA is consistent with it being a neurotransmitter in the cochlea. In addition the results: confirm the release of glutamate and taurine from the organ of Corti; suggest that additional substances may be released; and demonstrate the release of glutamate and taurine from tissue other than the organ of Corti.

An ipsilateral cochlear efferent loop protects the cochlea during intense sound exposure

One suggested physiological function of the efferent nerve fibers innervating the cochlea is that they protect the cochlea against the effects of intense sound exposure. In order to test this hypothesis, we studied the effects of intense sound in the presence and in the absence of strychnine which blocks the efferent nerve fibers. The results show that in presence of strychnine an ipsilateral intense sound has a greater effect on the cochlea than in the absence of strychnine. We conclude that the ipsilateral cochlear efferents may act as protectors against intense sound exposure.

Quisqualate excites spiral ganglion neurons of the guinea pig.

The effects of quisqualate on primary afferent auditory neurons were examined by comparing the unit activity of guinea pig spiral ganglion neurons before, during, and after the infusion of artificial perilymph containing 1 mM quisqualate into the basal turn scala tympani. In 10 of 13 preparations quisqualate infusion increased unit activity above baseline rates established prior to infusion while control infusions of quisqualate-free artificial perilymph had no appreciable influence on the unit activity of four preparations. Postexcitatory depression typically followed peak evoked excitation, but no purely inhibitory responses were observed. Postexcitatory depression developed into a temporary cessation of spike production in three cases, suggesting depolarization blockade had developed; however, all of these preparations gradually regained some degree of spontaneous unit activity following the termination of quisqualate infusion. Quisqualate-induced excitation may be attributable to the activation of receptors for the afferent neurotransmitter released by hair cells. This interpretation is consistent with our working hypothesis that the primary afferent neurotransmitter is L-glutamate or a structural analog of this excitatory amino acid.

Noise exposure alters the response of outer hair cells to ATP

The outer hair cells (OHCs) are one target of noise-induced effects. To date there are few studies which examine changes in the function of OHCs induced by noise exposure. There is increasing evidence that ATP may be a neuromodulator acting on OHCs. Therefore, we examined the possibility that the response to ATP may be altered by low-level noise exposure. ATP was tested on cation currents recorded from outer hair cells (OHCs) isolated from chronic noise-exposed guinea pigs and compared to currents recorded from normal control animals. The whole-cell variant of the patch-clamp technique was used. The incidence of response to 100 microM ATP was decreased in OHCs from noise-exposed animals as compared to controls when normal internal and external solutions were employed. When K+ was substituted by N-methyl-glucamine (NMG+) in the pipette solution, there were significant differences in the magnitudes of ATP-evoked currents between cells from noise-exposed and control animals. This was observed in both normal and 20 mM Ba2+ external solutions. In addition, the response to ATP exhibited a dependency on OHC length. In short OHCs (< 65 microns) from noise-exposed animals the magnitude of the response to ATP was significantly reduced. By contrast, the response in long OHCs (> 65 microns) from noise-exposed animals was increased. Results suggest that low-level noise exposure induces changes in OHCs which affect the response of the cell to ATP.

Volume regulation in cochlear outer hair cells

Many cells placed in a hypotonic medium initially swell and then rapidly undergo a regulatory volume decrease (RVD) to return towards original volume. Re-exposure to the isotonic solution results in the cells shrinking followed by a regulatory volume increase (RVI). Previous studies have shown that isolated outer hair cells (OHCs) placed in a hypotonic medium swell and maintain this shape until returned to the original medium. We re-examined this apparent lack of cell volume regulation in OHCs. OHCs were isolated from guinea pig cochleae, mechanically dissociated and dispersed, and placed in a Hanks balanced salt solution (HBS). In the cells studied, switching the perfusate to a hypotonic HBS (290-280 mmol/kg) for 15 min resulted in an immediate shortening of the OHCs (i.e., volume increase). In 26% of the cells, this increase was followed by a return to original length during the time the cell was perfused with the hypotonic medium, a RVD. Twelve percent of the cells demonstrating a RVD also displayed a RVI. Omitting collagenase and increasing Ca2+ concentration did not increase the percentage of cells displaying a RVD, while gadolinium (Gd3+, 10 microM) decreased the percentage to zero. This is the first report of isolated OHCs undergoing cell volume regulation.