Glutamate co-transmission from developing medial nucleus of the trapezoid body--lateral superior olive synapses is cochlear dependent in kanamycin-treated rats.

Cochlear dependency of glutamate co-transmission at the medial nucleus of the trapezoid body (MNTB)--the lateral superior olive (LSO) synapses was investigated using developing rats treated with high dose kanamycin. Rats were treated with kanamycin from postnatal day (P) 3 to P8. A scanning electron microscopic study on P9 demonstrated partial cochlear hair cell damage. A whole cell voltage clamp experiment demonstrated the increased glutamatergic portion of postsynaptic currents (PSCs) elicited by MNTB stimulation in P9-P11 kanamycin-treated rats. The enhanced VGLUT3 immunoreactivities (IRs) in kanamycin-treated rats and asymmetric VGLUT3 IRs in the LSO of unilaterally cochlear ablated rats supported the electrophysiologic data. Taken together, it is concluded that glutamate co-transmission is cochlear-dependent and enhanced glutamate co-transmission in kanamycin-treated rats is induced by partial cochlear damage.

Chronological changes in compromised olivocochlear activity and the effect of insulin in diabetic Wistar rats.

The aims of the present study were to investigate in diabetic rats: (1) the chronological changes of compromised medial olivocochlear bundle (MOCB) activity and auditory brainstem responses (ABR) and (2) the effect of insulin on diabetes-related hearing dysfunction. Diabetes mellitus was induced by intraperitoneal injection of streptozotocin. Thirty male Wistar rats were divided into three groups: control (C), diabetes with insulin injection (DI), and diabetes without insulin injection (DM). Click-evoked ABR, distortion product otoacoustic emission (DPOAE) and the contralateral suppression (CS) of DPOAE were measured for all animals monthly. Throughout the experiment, the thresholds of click-evoked ABR did not differ among groups. Wave III was delayed and interpeak latency I-III was prolonged in the DM group at the age of 29 weeks (p < 0.05). The amplitudes of the CS of DPOAE were markedly decreased after the 25th week in the DM group, but not in the C and DI groups. Compared to the C group, the CS in the DI group was not attenuated at any frequency. Dysfunction of auditory efferent olivocochlear activity developed in diabetic rats presenting no evidence of hearing loss. The finding of a significant decrease of the CS of DPOAE could be used as an earlier indicator of diabetes-related hearing impairment than changes of ABRs. The time course of compromised MOCB is positively correlated with the duration of diabetes. Insulin could therefore protect against compromised MOCB.

Slow build-up of cochlear suppression during sustained contralateral noise: central modulation of olivocochlear efferents?

The strength of the medial olivocochlear (OC) reflex is routinely assayed by measuring suppression of ipsilateral responses such as otoacoustic emissions (OAEs) by a brief contralateral noise, e.g., (Berlin, C.I., Hood, L.J., Cecola, P., Jackson, D.F., Szabo, P. 1993. Does type I afferent dysfunction reveal itself through lack of efferent suppression. Hear. Res. 65, 40-50). Here, we show in anesthetized guinea pigs, that the magnitude of OC-mediated suppression of ipsilateral cochlear responses (i.e., compound actions potentials (CAPs), distortion product (DP) OAEs and round-window noise) slowly builds over 2-3 min during a sustained contralateral noise. The magnitude of this build-up suppression was largest at low ipsilateral stimulus intensities, as seen for suppression measured at contra-noise onset. However, as a function of stimulus frequency, build-up suppression magnitude was complementary to onset suppression, i.e., largest at the lowest and highest frequencies tested. Both build-up and onset suppression were eliminated by cutting the OC bundle. In contrast to "slow effects" of shock-evoked medial OC activity (Sridhar, T.S., Liberman, M.C., Brown, M.C., Sewell, W.F. 1995. A novel cholinergic "slow effect" of efferent stimulation on cochlear potentials in the guinea pig. J. Neurosci. 15, 3667-3678), which are mediated by slow intracellular changes in Ca concentration in OHCs, build-up effects of contralateral noise are immediately extinguished upon OC bundle transection and are likely mediated by central modulation of the response rates in MOC fibers due to the sustained noise. Results suggest that conventional tests of OC reflex strength may underestimate its magnitude in noisy environments.

[Astragalus membranaceus promote expression of insulin-like growth factor 1 in rat model of olivo-cerebellar degeneration].

OBJECTIVE: To observe the effect of Astragalus membranaceus (AM) on insulin-like growth factor 1 (IGF-1) expression in a rat model of olivo-cerebellar degeneration and assess the neuroprotective actions of AM meanwhile. METHOD: Rats model of olivo-cerebellar degeneration was established by using 3-acetylpyridine. The effect of AM on the expression of Calbindin D-28K in inferior olive (IO) neurons by immunohistochemistry, the serum IGF-1 level by Elisa, the IGF-1 mRNA level in the cerebellum by RT-PCR were detected respectively. RESULT: AM effectively improve the serum IGF-1 level, Cerebellar IGF-1 mRNA level and the survival of the 10 neurons in a rat model of olivo-cerebellar degeneration, even at a lower dose (9 g x kg(-1)), and the effect was in a dose-dependent manner. CONCLUSION: AM could effectively upregulate the IGF-1 expression in the rat model of olivo-cerebellar degeneration, and have neuroprotective effect on IO neurons.

Neurotoxicity of BFM-95 on the medial olivocochlear bundle assessed by means of contralateral suppression of 2f1-f2 distortion product otoacoustic emissions.

OBJECTIVE: Berlin-Frankfurt-Munster 95 (BFM-95) is a common chemotherapeutic protocol against acute lymphoblastic leukemia (ALL). This prospective study investigates whether this protocol has an adverse effect on the medial olivocochlear bundle (MOCB) and/or outer hair cells' (OHCs) function. The distortion product otoacoustic emissions (DPOAEs) and their suppression by means of contralateral application of white noise were used for assessing the function of OHCs and the MOCB, respectively. STUDY DESIGN: Prospective study. SETTING: Oncology and otorhinolaryngology departments in a pediatric hospital. PATIENTS: Thirty-six children treated with ALL-BFM-95. INTERVENTIONS: Before chemotherapy, a baseline audiologic evaluation with tympanogram, standard and extended high frequency, pure-tone audiometry, and DPOAEs in the absence and presence of white noise was performed in all children. This population was divided in three groups. In a first group (n = 12), the evaluation was repeated after four sessions of vincristine administration; in the second group (n = 12), after 8 sessions; and in the third group (n = 12), several months after completion of the protocol. MAIN OUTCOME MEASURE: DPOAEs suppression by contralateral application of white noise. RESULTS: In the first and the third groups, we observed no changes in DPOAE amplitudes. Nevertheless, in the second group, the DPOAEs demonstrated significant decrease at 1416, 1685, 2002, and 2380 Hz. At baseline examination, all groups presented significant suppression at all frequencies. After eight vincristine sessions, instead of suppression, an increase of amplitudes was noted at 5 of 12 frequencies. Efferent-mediated DPOAE suppression reappeared in the third group at all frequencies (significant at 5 of 12 frequencies). CONCLUSION: ALL-BFM-95 seems to exert an early and reversible toxic effect on the MOCB, whereas its effects on OHCs are minimal and reversible. These minimal cochleotoxic and neurotoxic properties of ALL-BFM-95 might prove useful for research studies on the role of efferent innervation in hearing.

Neurotoxicity of vincristine on the medial olivocochlear bundle.

OBJECTIVE: Vincristine is a well known neurotoxic chemotherapeutic agent. Dose dependent and cumulative peripheral neuropathy is the main dose limiting side effect of chemotherapy with vincristine. The mechanisms responsible for the neurotoxic effects of vincristine have not yet been fully understood. This prospective study was directed at determining whether vincristine treatment interferes with the function of the medial olivocochlear bundle. DESIGN: Fifteen children suffering from leukemia were subjected to tympanogram, stapedial muscle reflex, pure tone audiometry and transient evoked otoacoustic emissions (TEOAEs) in the absence and presence of contralateral white noise on day 1 and on day 22 of treatment with vincristine. The function of the medial olivocochlear bundle was assessed by the phenomenon of suppression of otoacoustic emissions by contralateral application of white noise. RESULTS: The study revealed a statistically significant decrease of contralateral suppression amplitudes in all cases after three sessions of chemotherapy with vincristine. On the contrary no alterations were observed in pure tone audiometry thresholds. A non-significant decrease of the mean TEOAEs' amplitudes was also noted. When analyzed by frequency, however, this decrease reached the level of statistical significance at two frequencies. CONCLUSION: Vincristine treatment seems to exert a neurotoxic effect on the efferent olivocochlear system, which takes place early in the course of chemotherapy. This is a new aspect to be added to the possible mechanisms underlying the toxicity of vincristine in the auditory periphery. Whether changes in efferent function might contribute to understanding the mechanisms of neurotoxicity caused by vincristine, or find any clinical application as a predictor or early detector of neurological side effects of vincristine still remains to be seen.

Baclofen attenuates harmaline induced tremors in rats.

Recent experimental and clinical studies clearly suggest the role of gamma-aminobutyric acid (GABA) in the pathogenesis of tremors. The present study was undertaken to investigate the effect of baclofen, a GABA B receptor agonist on harmaline induced tremors. Four groups of female Wistar rats weighing 100+/-15 g were injected with harmaline (10 mg/kg, intraperitoneally) for inducing experimental tremors. The animals in groups 2, 3 and 4 were given baclofen by gavage at doses of 2.5, 5 and 10 mg/kg, respectively, half an hour before harmaline administration, whereas, the rats in group 1 served as control and received water. The latency of onset, intensity and duration of tremor and electromyographic (EMG) responses were recorded. Treatment with baclofen resulted in a dose dependent decrease in the intensity of tremor. Our EMG study also revealed a significant decrease in the amplitude of tremors in baclofen treated rats. A highly significant increase in latency of onset of tremor was observed in the rats treated with high dose (10 mg/kg) of baclofen only. This study clearly suggests beneficial effects of baclofen in harmaline induced tremors.

Beta-endorphin- and GABA-mediated depressor effect of specific electroacupuncture surpasses pressor response of emotional circuit.

It has been proved that input of specific electroacupuncture (EA) can activate beta-endorphin(beta-EP)ergic and noradrenergic neurons projecting to the rostral ventrolateral medulla (RVL), the latter acting upon the RVL-GABAergic interneurons, thereby produce depressor effect. The present study further shows that: (1) The EA depressor effect is strong enough to surpass the pressor response of the AC (nucleus amygdaloideus centralis)-emotional circuit, (2) both beta-endorphin (beta-EP) and GABA in the RVL mediate the EA antagonistic effect, (3) the EA effect does not take place in the AC and paraventricular nucleus (two key nuclei besides the RVL, which also have beta-EPergic input) in the emotional circuit.

Organization of olivocerebellar activity in the absence of excitatory glutamatergic input.

The olivocerebellar system has been proposed to function as a timing device for motor coordination in which inferior olivary neurons act as coupled oscillators that spontaneously generate rhythmic and synchronous activity. However, the inferior olive receives excitatory afferents, which can also drive the activity of these neurons. The extent to which the olivocerebellar system can intrinsically generate synchronous activity and olivary neurons act as neuronal oscillators has not been determined. To investigate this issue, multiple electrode recordings of complex spike (CS) activity were obtained from 236 crus 2a Purkinje cells in anesthetized rats. Intraolivary injections of the glutamate antagonists 6-cyano-7-nitroquinoxaline-2,3-dione or 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium were made, and the resulting changes in CS activity were determined. Loss of evoked CS responses to motor cortex stimulation or perioral tactile stimulation was used to measure the efficacy of the block. Block of glutamatergic input decreased the average CS firing rate by approximately 50% but did not abolish spontaneous CS activity. The remaining CS activity was significantly more rhythmic than that in control. The patterns of synchrony were similar to those found in control conditions (i.e., synchronous CSs primarily occurred among Purkinje cells located within the same approximately 250-microm-wide rostrocaudally oriented cortical strip); however, this normal banding pattern was enhanced. These changes in CS activity were not observed with vehicle injections. The results suggest that excitatory afferent activity disrupts olivary oscillations and support the hypotheses that olivary neurons are capable of acting as neuronal oscillators and that synchronous CS activity results from electrotonic coupling of olivary neurons.

Effects of (-)-baclofen, clonazepam, and diazepam on tone exposure-induced hyperexcitability of the inferior colliculus in the rat: possible therapeutic implications for pharmacological management of tinnitus and hyperacusis.

Recent investigations in the authors' laboratory have shown that acute tone exposure (4 kHz continuous tone, 104 dB sound pressure level (SPL), 30-min duration) induces increases in the amplitude of click-evoked potentials in the inferior colliculus (IC). These increases have been attributed to a decrease in GABAA-mediated inhibition on IC neurons. In the present study, we examined the effects of three compounds (diazepam, clonazepam, and (-)-baclofen) that are known to enhance GABAergic inhibition on these tone exposure-induced increases and on changes in temporal integration in the IC. (-)-Baclofen was the only one of the three compounds tested that reversed in a dose-dependent manner the effects of tone exposure on both the amplitude of the click-evoked potentials recorded from the IC and on measures of the changes in temporal integration based on these potentials. Diazepam and clonazepam exhibited remarkably different effects on the click-evoked potentials recorded from the surface of the IC. Diazepam caused a dose-dependent decrease in one of the components of the IC potentials that reflects postsynaptic activity in the IC, whereas clonazepam caused a dose-dependent decrease in a peak that reflects input to the IC from the superior olivary complex (SOC). At dosages up to 40 mg/kg, neither diazepam nor clonazepam reversed the changes in temporal integration in the IC that were induced by the tone exposure; diazepam caused a small, but statistically significant, enhancement of the effects of tone exposure on this function. The results of this study show that (-)-baclofen is a potent modulator of both the excitability of neurons in the ascending auditory pathway and the processing of auditory information by IC neurons. The finding of the present study that two benzodiazepines (clonazepam and diazepam) have remarkably different effects on evoked potentials, which reflects both input to the IC and postsynaptic events in the IC neurons, suggests heterogenicity of the GABAA receptor from one structure to another in the ascending auditory pathway. We suggest that (-)-baclofen may be clinically useful in treating disorders of the auditory system that are caused by plasticity in the ascending auditory pathway.

Generators of the brainstem auditory evoked potential in cat. II. Correlating lesion sites with waveform changes.

Brainstem regions involved in generating the brainstem auditory evoked potential (BAEP) were identified by examining the effects of lesions on the click-evoked BAEP in cats. An excitotoxin, kainic acid, was injected into various parts of the cochlear nucleus (CN) or into the superior olivary complex (SOC). The locations of the resulting lesions were correlated with the changes produced in the various extrema of the BAEP waveforms. The results indicate that: (1) the earliest BAEP extrema (P1, N1 (recorded between vertex and the earbar ipsilateral to the stimulus) and P1a, P1b, (vertex to contralateral earbar)) are generated by cells with somata peripheral to the CN; (2) P2 is primarily generated by posterior anteroventral CN (AVCNp) and anterior posteroventral CN (PVCNa) cells; (3) SOC, anterior anteroventral CN (AVCNa), AVCNp, and PVCNa cells are involved in generating P3; (4) AVCNa cells are the main CN cells involved in P4, N4, and P5 generation; (5) both ipsilateral and contralateral SOC cells have a role in generating monaurally evoked P4 and P5; and (6) P5 is generated by cells with characteristic frequencies below 10 kHz. From (2) and (4), it is clear that P2 and P4-P5 are generated by cells in distinct, parallel pathways.

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

Time-varying alterations in the f2-f1 DPOAE response to continuous primary stimulation. I: Response characterization and contribution of the olivocochlear efferents.

The f2-f1 distortion product otoacoustic emission (DPOAE) can be observed to undergo gradual alterations in amplitude during continuous ipsilateral stimulation with primary tones. In the present experiments, we characterized the dependence of these amplitude alterations on several stimulus variables (intensity, duration, frequency) and on DPOAE type (quadratic vs cubic) and tested the hypothesis that such alterations are mediated by the olivocochlear (OC) efferents. Responses were recorded in urethane-anesthetized guinea pigs with sectioned middle ear muscles before and after intracochlear application of antagonists (curare, 1 microM; bicuculline, 10 microM; tetrodotoxin, 1 microM) or before and after OC efferent section at the midline of the floor of the IVth ventricle. We confirm previous reports of continuous stimulation-related alterations in the amplitude of the quadratic distortion product, f2-f1, and report a novel, suppressive 'off-effect' apparent in f2-f1 amplitude following a short rest from such stimulation. Response alterations were sensitive to primary intensity and to duration of rest from continuous stimulation, but were not clearly frequency-dependent over the ranges tested. Corresponding alterations in the amplitude of the cubic nonlinearity, 2f1-f2 were very small or absent. Amplitude alterations in f2-f1 were reduced but not blocked by OC efferent antagonists (curare, bicuculline) and were largely unaffected by TTX or by midline brainstem section. All of these manipulations, however prevented completely the known efferent-mediated contralateral sound suppression of both f2-f1 and 2f1-f2 DPOAEs. Taken together, these results do not provide support for efferent control of the f2-f1 amplitude alterations observed during continuous ipsilateral stimulation.

Lesions of the inferior olive do not affect long- or short-term habituation of the acoustic startle response in rats.

Short- and long-term habituation of the acoustic startle response were assessed in a group of inferior olive-lesioned rats. Neither short- and long-term habituation, nor the performance of the reflex, were affected by the lesion. Since the cerebellar vermis is essential for long-term habituation of this reflex, we suggest that climbing fibres are not involved in this form of learning, which would therefore be mediated by the other cerebellar input, presumably the mossy fibres.

Ethanol increases single unit activity in the inferior olivary nucleus.

Single unit recording of rat inferior olivary nucleus neurons reveals significantly elevated discharge after acute intraperitoneal injection of 2 g/kg ethanol. This effect is consistent across 3 different methods of anesthesia and immobilization: local Xylocaine plus intraperitoneal D-tubocurare, intraperitoneal chloral hydrate and halothane vapor. In contrast, under urethane anesthesia acute ethanol produces significant depression of olivary discharge. Since this effect is opposite to that found under the other anesthetic conditions (including topical Xylocaine only), urethane anesthesia may compromise generalizations of electrophysiologic studies of ethanol. Neurons of the inferior olivary nucleus excite cerebellar Purkinje cells through a powerful afferent circuit; our data therefore suggest that ethanol-induced increases in cerebellar Purkinje cell complex (climbing fiber burst) spikes, obtained in our previous studies, are secondary to olivary activation.

[Opiate receptors and sleep. II. Effects of micro-injection of ethyl alcohol and pentobarbital in the median thalamus, periaqueductal gray matter and nucleus of the tractus solitarius of the rabbit (author's transl)]. / Récepteurs de l'opium et sommeil. II. Effets de micro-injections d'alcool éthylique et de pentobarbital dans le thalamus médian, la substancae grise centrale périaqueducale et du noyau du tractus solitaire du lapin.

There is an analogy between sleep EEGs produced by microinjections of morphine in the bulbo-mesencephalo-thalamic recruiting system and EEGs seen during anesthetic-induced sleep. Many studies in the last 10 years have claimed cross-tolerance and cross-dependence between opiates and ethyl alcohol. Opiates, ethyl alcohol and pentobarbital have many common metabolic actions in the central nervous system. Like morphine, microinjections of optimal equimolar doses of ethyl alcohol and pentobarbital in the bulbo-mesencephalo-thalamic sleep-inducing system of the rabbit produce sleep EEGs with abundant fast activity, which is blocked by naloxone (2 mg/kg i.v.) or by microinjections (160 micrograms) into the same structures. However, there is neither binding nor displacement by naloxone of ethyl alcohol or pentobarbital from the opiate receptor. It is thus probable that, via an as yet unknown mechanism, ethyl alcohol and pentobarbital promote the release of endorphins or peptides, which are specific ligands of all or some opiate receptors.