P2X receptor-mediated changes in cochlear potentials arising from exogenous adenosine 5'-triphosphate in endolymph

Our previous studies have determined the presence of adenosine 5'-triphosphate (ATP) in the cochlear fluids and shown that extracellular ATP introduced into the endolymphatic compartment of the guinea pig cochlea has a significant dose-dependent suppressive effect on both endocochlear potential (EP) and cochlear microphonic (CM), which is mediated via P2 receptors. In the present study, the influence of P2 receptor agonists and antagonists on this suppressive effect was investigated to characterise the subtypes of P2 receptor mediating the ATP-induced effect on cochlear function. Using a double-barreled pipette attached to a pressure injector, small volumes (2-10 nl) of ATP (0.01-1 mM) and P2 receptor agonists or P2 receptor antagonists in artificial endolymph were introduced into the scala media of the first (basal) and third turns of the guinea pig cochlea, while the EP and CM were monitored. ATP and P2 receptor agonists (5x10(-14)-1x10(-11)cibacron blue. Neither adenosine nor uridine 5'-triphosphate (2x10(-13)-2x10(-11) moles) nor the P2 receptor antagonists on their own had any effect on EP and CM. The ATP effect on the potentials was greater at the third cochlear turn when compared to the first turn. These results provide evidence that in the endolymphatic compartment of the guinea pig, the extracellular ATP effect on cochlear function is likely mediated through an interaction with P2 receptors which assemble as ATP-gated ion channels.     

Measurement of guinea pig basilar membrane using computer-aided three-dimensional reconstruction system

Cochleas are known to have the ability to analyze a frequency widely, and this ability seems to be owed mostly to the basilar membrane (BM) configuration. However, the relationship between the cochlear frequency-position map and the BM configuration is not clear. Therefore, in this paper, the internal structures of a guinea pig cochlea, especially the BM configuration, were reconstructed and measured using a computer-aided three-dimensional (3-D) reconstruction system. Then, an attempt was made to examine the influence of the BM configuration on the cochlear frequency-position map. The measurement results indicate that the width of the BM increased and its thickness decreased with an increase in the distance from the basal turn towards the apical turn. Theoretical consideration reveals that the wide frequency-position of the cochlea is achieved by not only the BM configuration change along the length of the cochlea but also the change of the Young's modulus of the BM along the length of the cochlea.     

Nitric oxide in glutamate-induced compound action potential threshold shifts

OBJECTIVE: Investigate the role of NO as a neurotransmitter in the gerbil cochlea and the effects of (7-NI) on compound action potential (CAP) threshold elevations induced by l-glutamate, an agonist at the NMDA glutamate receptor subtype, to further elucidate the role of NO in cochlear excitotoxicity. METHOD: In anesthetized gerbils, CAP thresholds were recorded before and after cochlear perfusions with a control solution of artificial perilymph (APS) and a test solution of L-glutamate (GA) in three experimental groups. RESULTS: The control group showed no CAP threshold elevations (p<0.05) when APS was perfused after systemic pre-treatment with 7-NI. GA perfusion alone caused significant elevation (p<0.05) of the mean cochlear CAP threshold (25 dB SPL+/-5.8 dB to 78 dB SPL+/-19.5 dB). The CAP threshold elevation was prevented (p<0.05) when the animals were pretreated with 7-NI before GA perfusion (24 dB SPL+/-4.2dB to 27 dB SPL+/-6.7 dB). CONCLUSION: NO mediates excitotoxicity when the cochlea is perfused with L-glutamate.     

Effects of lidocaine on basilar membrane vibration in the guinea pig.

The effects of lidocaine on basilar membrane (BM) vibration and compound action potential (CAP) were studied in guinea pigs in order to elucidate the site of lidocaine action in the cochlea. BM vibration was measured with a laser Doppler vibrometer through an opening made in the lateral bony wall of the scala tympani at the basal turn. Ten min after local administration of lidocaine (250 microg) into the scala tympani, the velocity of BM vibration and the CAP amplitude decreased significantly at around the characteristic frequency of the stimulus sound (p < 0.05). The maximum decreases were 4 dB in the velocity of the BM vibration and 40 dB in the CAP amplitude. In contrast, such changes were not observed after i.v. injection of lidocaine (1.5 mg/kg). These results suggest that when lidocaine is administered locally in the cochlea it acts not only on the cochlear nerve but also on the outer hair cells.     

Effects of Lidocaine on Basilar Membrane Vibration in the Guinea Pig

The effects of lidocaine on basilar membrane (BM) vibration and compound action potential (CAP) were studied in guinea pigs in order to elucidate the site of lidocaine action in the cochlea. BM vibration was measured with a laser Doppler vibrometer through an opening made in the lateral bony wall of the scala tympani at the basal turn. Ten min after local administration of lidocaine (250 &#119 g) into the scala tympani, the velocity of BM vibration and the CAP amplitude decreased significantly at around the characteristic frequency of the stimulus sound ( p <0.05). The maximum decreases were 4 dB in the velocity of the BM vibration and 40 dB in the CAP amplitude. In contrast, such changes were not observed after i.v. injection of lidocaine (1.5 mg kg). These results suggest that when lidocaine is administered locally in the cochlea it acts not only on the cochlear nerve but also on the outer hair cells.     

Spiral ligament and stria vascularis changes in cochlear otosclerosis: effect on hearing level.

OBJECTIVE: To investigate the effect of changes within the spiral ligament and stria vascularis on hearing in cochlear otosclerosis, we examined spiral ligament hyalinization, stria vascularis atrophy, and sensory hearing loss in cochlear otosclerosis and described changes in ion transport molecule expression. STUDY DESIGN: Retrospective. SETTING: Tertiary referral center. PATIENTS: Thirty-two cochleae from 24 temporal bone donors with histologic evidence of cochlear otosclerosis, including spiral ligament hyalinization. INTERVENTION: Audiography. MAIN OUTCOME MEASURES: Measurements of spiral ligament width, stria vascularis, and bone-conduction thresholds were compared by the amount of hyalinization. Expression of the ion transport molecules Na,K-ATPase, connexin 26, and carbonic anhydrase II were assessed by immunohistochemical techniques. RESULTS: Hyalinization most often involved the posterior basal turn (88%) and the posterior middle turn (27%). Spiral ligament hyalinization correlated significantly with stria vascularis atrophy in the posterior middle turn of the cochlea (rho = -0.63, p < 0.01). There was a trend toward a significant association in the posterior basal turn (rho = -0.31, p < 0.08). Bone-conduction thresholds at 2,000 and 4,000 Hz were significantly associated with the amount of stria vascularis atrophy (rho = -0.44, -0.40, p < 0.05). In addition, we observed decreased immunostaining for both carbonic anhydrase II with Type I fibrocytes and Na,K-ATPase with stria vascularis and Type II and Type IV fibrocytes of the spiral ligament in cochlear otosclerosis sections compared with normal cochlea. Na,K-ATPase staining within the stria vascularis was further decreased in the presence of spiral ligament hyalinization. No significant differences were seen with connexin 26 immunostaining. However, immunostaining results were somewhat inconsistent. CONCLUSION: These data suggest that spiral ligament structure and function are essential for stria vascularis survival. In addition, dampened expression of ion transport molecules within the spiral ligament and stria vascularis may disrupt potassium ion recycling, resulting in loss of endocochlear potential and sensory hearing loss.     

Combined effects of acute lead acetate exposure and tone exposure of the guinea pig cochlea

Lead acetate exposure to humans can induce various disorders of the cranial nerves. Although vertigo and sensorineural deafness have been reported in lead workers, the dose effects of lead acetate on the cochlea and eighth cranial nerve are not well documented. We investigated the effects of lead acetate on the male albino Hartley guinea pig cochlea by measuring cochlear microphonics (CM), whole nerve action potential (AP), endocochlear potential (EP) and K⁺ ion concentration of the endolymph. Animals were given lead acetate by intraperitoneal injection as 20 mg/week for 4 consecutive weeks. A total dose < 80 mg did not induce electrophysiological changes in the cochlea. However, the AP output voltage (N₁) decreased if the 80 mg lead acetate treatment was followed by an 80 dB tone exposure at 6 kHz during 24 h. A change was observed in CM and EP but not K⁺ ion concentration in the scala media.     

Distribution of immunophilin FKBP-12 protein and mRNA within the mammalian cochlea and cochlear nucleus

Immunophilin FK binding protein-12 (FKBP-12), the soluble receptor for the immunosuppressant drug FK506, is involved in a number of neuronal activities including increased nerve regeneration in the peripheral nervous system and enhanced recovery in animal models of neurodegenerative diseases. In addition, FKBP-12 is tightly bound to the calcium release channel ryanodine receptor and physiologically interacts with the inositol 1,4,5-trisphosphate receptor. In nearly all cell types, release of intracellular Ca(2+) and subsequent second messenger signaling involves activation of these ion channels. We determined the distribution of FKBP-12 within the mammalian cochlea and dorsal cochlear nucleus (DCN) in order to gain insight into Ca(2+) regulation within the cochlea and to possibly identify potential cellular targets for neuroimmunophilin ligands that may prove useful in protection and recovery following ototoxic insult. FKBP-12 protein and mRNA were found to be abundant throughout rat and guinea pig cochlea and DCN.     

Purinergic regulation of sound transduction and auditory neurotransmission

In the cochlea, extracellular ATP influences the endocochlear potential, micromechanics, and neurotransmission via P2 receptors. Evidence for this arises from studies demonstrating widespread expression of ATP-gated ion channels (assembled from P2X receptor subunits) and G protein-coupled receptors (P2Y receptors). P2X2 receptor subunits are localized to the luminal membranes of epithelial cells and hair cells lining scala media. These ion channels provide a shunt pathway for K+ ion egress. Thus, when noise exposure elevates ATP levels in this cochlear compartment, the K+ conductance through P2X receptors reduces the endocochlear potential. ATP-mediated K+ efflux from scala media is complemented by a P2Y receptor G protein-coupled pathway that provides coincident reduction of K+ transport into scala media from the stria vascularis when autocrine or paracrine ATP signalling is invoked. This purinergic signalling likely provides a basis for a reactive homoeostatic regulatory mechanism limiting cochlear sensitivity under stressor conditions. Elevation of ATP in the perilymphatic compartment under such conditions is also likely to invoke purinergic receptor-mediated changes in supporting cell micromechanics, mediated by Ca2+ influx and gating of Ca2+ stores. Independent of these humoral actions, ATP can be classified as a putative auditory neurotransmitter based on the localization of P2X receptors at the spiral ganglion neuron-hair cell synapse, and functional verification of ATP-gated currents in spiral ganglion neurons in situ. Expression of P2X receptors by type II spiral ganglion neurons supports a role for ATP as a transmitter encoding the dynamic state of the cochlear amplifier.     

Ion flow in cochlear hair cells and the regulation of hearing sensitivity

This paper discusses how ion transport proteins in the hair cells of the mammalian cochlea work to produce a sensitive but stable hearing organ. The transport proteins in the inner and outer hair cells are summarized (including their current voltage characteristics), and the roles of these proteins in determining intracellular Ca(2+), membrane potential, and ultimately cochlear sensitivity are discussed. The paper also discusses the role of the Ca(2+) sequestration sacs in outer hair cells in the autoregulation of hair cell membrane potential and cochlear gain, and how the underdamped control of Ca(2+) within these sacs may produce the observed slow oscillations in cochlear sensitivity and otoacoustic emissions after cochlear perturbations, including perilymphatic perfusions and prolonged low-frequency tones. The relative insensitivity of cochlear gain to short-term changes in the endocochlear potential is also discussed.     

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.     

The time-course of furosemide-induced strial changes in guinea pigs after pretreatment with daltroban

It was shown previously (Ernst et al., 1989) that pretreatment of guinea pigs with a thromboxane (TX) receptor antagonist attenuates the decline of the endocochlear potential (EP) induced by furosemide. The present paper is aimed at investigating a possible correlation between the electrophysiological data and ultrastructural changes of the stria vascularis by electron microscopy. The dosages of 40, 60, and 80 mg/kg furosemide were injected after the pretreatment with the TX receptor antagonist daltroban and compared to controls which were injected with furosemide only. It was found at all furosemide concentrations that the strial changes 10 min after injection were nearly unchanged against controls. 30 min after furosemide injection, the most pronounced changes were seen when pretreating the animals: a clear reduction of the marginal cell swelling and edema in general were observed at 40 and 60 mg/kg furosemide. The guinea pigs injected with 80 mg/kg furosemide after pretreatment displayed nearly the same changes as controls.     

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.     

Receptor maturation and synaptogenesis in the golden hamster cochlea

Maturation of hamster cochlea was studied using light and electron microscopy. Critical stages of receptor and neural structure development have been determined. At birth the hamster cochlea shows a pronounced immaturity, but innervation can already be found. 2 or 3 days later, characteristic afferent synapses can be recognized beginning at the inner hair cell level. Similarly, efferent endings first appear on the inner side at the end of the first week. The onset of auditory function must be related to structures depicted at around 10 days, and cochlear maturation is achieved at about 25 days. The sequence of synaptic development in the cochlea is discussed regarding the general morphogenesis of synapses within the nervous system. Some determinations of the timing of peripheral myelination are given. This process begins almost a week before the presumable date of the onset of function.     

Role of thromboxane A2 in a microcirculation disorder of the rat inner ear

Summary Since thromboxane (TX) A₂ causes vasoconstriction and platelet aggregation, we evaluation the effect of a TXA₂ receptor antagonist (vapiprost) and a TXAZ synthetase inhibitor (Y-20811) on a microcirculation disorder in the rat inner ear that was induced by a photochemical reaction between an intravenous injection of rose bengal (RB) and green light. A gradual decrease of the cochlear action potential (CAP) to an 8 kHz sound stimulus was measured with an electrocochleogram and occurred after the RB injection. The CAP then disappeared 5 min after the injection of RB. Both vapiprost and Y-20811 significantly prolonged the time required to complete suppression of the CAP as compared with saline as control. These findings indicate that TXAZ may play an important role in microcirculation disorders in the rat inner ear.     

Absence of mRNA encoding estrogen receptor in the rat cochlea

Based on changes in hearing thresholds and tinnitus that are co-related with the menstrual cycle, it has been suggested that the cochlea may respond directly to estrogen. For this to occur, the cochlea should express estrogen receptors. In situ mRNA hybridization was performed on normal female rat cochleas, using radiolabeled RNA probes complementary to mRNA encoding estrogen receptor, to determine whether estrogen receptors are present in the cochlea. Strong hybridization of the riboprobes to sections of uterus and hypothalamus indicated that the technique detected estrogen receptor mRNA. No hybridization to any cochlear tissues was observed. The results indicate that estrogen receptors are not expressed on cochlear cells, at least in rats. This in turn suggests that variation in cochlear responses during the estrus cycle are not the result of the direct effect of estrogen on the cochlea. Such variation may, however, be caused by systemic changes in fluid regulation induced by estrogen receptors at a distant site, or by other hormone receptors.     

Electrical resistivity measurements in the mammalian cochlea after neural degeneration

OBJECTIVES/HYPOTHESIS: In the present series of experiments, the effect of neural degeneration on the cochlear structure electrical resistivities was evaluated to test if it alters the current flow in the cochlea and if increased current levels are needed to stimulate the impaired cochlea. In cochlear implants, frequency information is encoded in part by stimulating discrete populations of spiral ganglion cells along the cochlea. However, electrical properties of the cochlear structures result in shunting of the current away from the auditory neurons. This consumes energy, makes cochlear implants less efficient, and drastically reduces battery life. Models of the electrically stimulated cochlea serve to make predictions on current paths using modified and improved cochlear implant electrodes. However, one of the model's shortcomings is that most of the values for tissue impedances are not direct measurements. They are derived from bulk impedance measurements, which are fitted to lumped-element models. STUDY DESIGN: The four-electrode reflection-coefficient technique was used to measure resistivities in the gerbil cochlea. In vivo and in vitro (the hemicochlea) models were used. Measurements were made in normal and in deafened animals. Cochlear damage was induced by neomycin injection into the animals' middle ears. Neural degeneration was allowed to occur over 2 months before performing the measurements in the deafened animals. RESULTS: The resistivity values in deafened animals were smaller than in the normal-hearing animals, thus altering the current flow within the cochlea. CONCLUSIONS: Resistivity changes and subsequent changes in current path should be considered in future designs of cochlear implants.     

The cochlear amplifier: augmentation of the traveling wave within the inner ear

PURPOSE OF REVIEW: There have been many recent advancements in our understanding of cochlear function within the past ten years. In particular, several mechanisms that underlie the sensitivity and sharpness of mammalian tuning have been discovered. This review focuses on these issues. RECENT FINDINGS: The cochlear amplifier is essentially a positive feedback loop within the cochlea that amplifies the traveling wave. Thus, vibrations within the organ of Corti are sensed and then force is generated in synchrony to increase the vibrations. Mechanisms that generate force within the cochlea include outer hair cell electromotility and stereociliary active bundle movements. These processes can be modulated by the intracellular ionic composition, the lipid constituents of the outer hair cell plasma membrane, and the structure of the outer hair cell cytoskeleton. SUMMARY: A thorough understanding of the cochlear amplifier has tremendous implications to improve human hearing. Sensorineural hearing loss is a common clinical problem and a common site of initial pathology is the outer hair cell. Loss of outer hair cells causes loss of the cochlear amplifier, resulting in progressive sensorineural hearing loss.     

Narrow-band action potentials of the guinea-pig cochlea as compared with the ordinary electrocochleograms under normal and pathological conditions

The narrow-band action potentials (NAP) and the ordinary electrocochleograms were recorded from the guinea-pig cochlea under normal and pathological conditions in order to study whether the NAP could be a useful measure to detect cochlear dysfunctions. The cochlea damaged either by the administration of kanamycin or by a mechanical lesion of the round window served as pathological materials. Recordings showed that the threshold and amplitude measured for the N1 potential of NAPs ran in parallel with those of the cochlear microphonic potentials (CM), under both normal and pathological conditions of the cochlea. This implies that the CM could be replaced by the NAP when difficulties were present in recording CMs. It may be inferred that the NAP reflects responses of the inner hair-cells and cochlear nerves, while the CM would mainly be derived from responses of the outer hair-cells to the frequency-specific movements of the basilar membrane. If so, the NAP should offer a good means for the objective audiometry. Recording NAPs is also superior to the ordinary electrocochleography in that the method makes it possible to obtain responses generated near the apex of the cochlea, i.e., responses to low-pitch sound stimuli.