Sanford C. Bledsoe

Descending projections to the dorsal and ventral divisions of the cochlear nucleus in guinea pig

The origins of extrinsic projections to the guinea pig dorsal and ventral cochlear nuclei were identified by examining the retrograde transport of horseradish peroxidase conjugated to wheatgerm agglutinin following its injection into each of these divisions. Major projections originated in periolivary regions of the superior olivary complex, the contralateral cochlear nucleus and the inferior colliculus. There was no contribution from the nuclei of the lateral lemniscus to these pathways. The heaviest projection from the periolivary regions to both divisions of the cochlear nucleus arose bilaterally in the ventral nucleus of the trapezoid body. The ipsilateral lateral nucleus of the trapezoid body also projected heavily to dorsal and ventral cochlear nucleus. In addition, the ventral cochlear nucleus received a substantial projection from the dorsal aspect of the ipsilateral dorsomedial periolivary nucleus. Projections originating bilaterally in the central nucleus of the inferior colliculus terminated in the deep layers of dorsal cochlear nucleus. These projections appear to be more strongly ipsilateral and specific than those reported in the cat.

Immunocytochemical and lesion studies support the hypothesis that the projection from the medial nucleus of the trapezoid body to the lateral superior olive is glycinergic

Pre- and postembedding immunocytochemical techniques were used to study the distribution of glycine immunoreactivity in the superior olivary complex of guinea pigs following kainic acid (KA) lesions of the medial nucleus of the trapezoid body (MNTB). Destruction of the MNTB by injecting 50-100 nl of 10 mM KA virtually abolished labeled neurons in the MNTB at the site of the lesion. This resulted in a marked decrease in the number of labeled fibers projecting to the ipsilateral lateral superior olive (LSO) and in the number of labeled fibers and presynaptic terminals in the neuropil of the LSO. Smaller volumes (20 nl) of KA produced similar but more restricted changes that conformed to the topographic organization of the MNTB projection to the LSO. The results support the hypothesis that the MNTB to LSO pathway is glycinergic.

Disruption of Lateral Efferent Pathways: Functional Changes in Auditory Evoked Responses

The functional consequences of selectively lesioning the lateral olivocochlear efferent system in guinea pigs were studied. The lateral superior olive (LSO) contains the cell bodies of lateral olivocochlear neurons. Melittin, a cytotoxic chemical, was injected into the brain stem using stereotaxic coordinates and near-field evoked potentials to target the LSO. Brain stem histology revealed discrete damage to the LSO following the injections. Functional consequences of this damage were reflected in depressed amplitude of the compound action potential of the eighth nerve (CAP) following the lesion. Threshold sensitivity and N1 latencies were relatively unchanged. Onset adaptation of the cubic distortion product otoacoustic emission (DPOAE) was evident, suggesting a reasonably intact medial efferent system. The present results provide the first report of functional changes induced by isolated manipulation of the lateral efferent pathway. They also confirm the suggestion that changes in single-unit auditory nerve activity after cutting the olivocochlear bundle are probably a consequence of disrupting the more lateral of the two olivocochlear efferent pathways.

Inhibitory Amino Acid Synapses and Pathways in the Ventral Cochlear Nucleus

The processing of auditory information requires neurons to fire at a rapid rate. Hair cells can, for example, respond to tones by oscillations of membrane potential at rates as high as many hundred times per second. This response rate firing conveys important information about intensity and frequency and so is maintained throughout the initial processing in the auditory brain stem. The first synapse through which response rate information must be accurately passed is between the inner hair cells and the auditory nerve. It is not surprising that evidence supports an excitatory amino acid transmitter and an ionotropic excitatory amino acid receptor at this synapse (Bledsoe et al.,’ 88; Altschuler et al.,’ 89), since this is one of the few known excitatory transmitter — receptor combinations capable of such speed. The next synapse in the ascending auditory pathway is obligatory, in the cochlear nucleus (CN). Firing rate information transfer is maintained at many auditory nerve — CN synapses, leading to the expectation that these synapses would also have an excitatory transmitter — receptor combination capable of considerable speed. Much evidence suggests that an excitatory amino acid is the auditory nerve transmitter (chapter in this volume by Juiz et al.; Wenthold,’ 91 and Caspary,’ 86) and that most CN projection neurons receive this input via an ionotropic excitatory amino acid receptor (see Wenthold et al. and Morest, chapters this volume), currently termed a glutamate receptor (GluR).

A shuttered neural probe with on-chip flowmeters for chronic in vivo drug delivery

This paper describes the development of shutters, fluidic ribbon cables, and flowmeters for use on bulk-micromachined neural probes. The resulting devices permit electrical recording, stimulation, and chemical drug delivery in the central nervous system at the cellular level. Dielectric shutters reduce unintended drug delivery by a factor of about 25 compared with an open orifice, while silicon fluidic cables are 1.5 times more flexible than the 70 mum-ID polyimide tubing previously used. A pulsed thermal flowmeter integrated on the probe allows verification of drug delivery on a per-channel basis with a resolution of 150 pL/s. Thermal time constants are about 60 mus, allowing multiple measurements within one delivery pulse while restricting any tissue heating to negligible levels. Finally, test structures have shown the process compatibility of on-chip thermopneumatic microvalves and micropumps. The fabrication of these fluidic structures (microchannels, shutters, cables, valves, and pumps) requires only two masks in addition to the eight normally used for passive probes, providing important new capabilities for use in neurophysiological research and neural prostheses

Disruption of Lateral Olivocochlear Neurons via a Dopaminergic Neurotoxin Depresses Sound-Evoked Auditory Nerve Activity

We applied the dopaminergic (DA) neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to the guinea pig cochlear perilymph. Immunolabeling of lateral olivocochlear (LOC) neurons using antibodies against synaptophysin was reduced after the MPTP treatment. In contrast, labeling of the medial olivocochlear innervation remained intact. As after brainstem lesions of the lateral superior olive (LSO), the site of origin of the LOC neurons, the main effect of disrupting LOC innervation of the cochlea via MPTP was a depression of the amplitude of the compound action potential (CAP). CAP amplitude depression was similar to that produced by LSO lesions. Latency of the N1 component of the CAP, and distortion product otoacoustic emission amplitude and adaptation were unchanged by the MPTP treatment. This technique for selectively lesioning descending LOC efferents provides a new opportunity for examining LOC modulation of afferent activity and behavioral measures of perception.

Comparative actions of salicylate on the amphibian lateral line and guinea pig cochlea

1. Salicylate actions on afferent nerve activity in the Xenopus lateral line and on cochlear potentials in guinea pig were investigated. 2. In the lateral line, salicylate (0.3-2.5 mM) suppressed spontaneous activity, water motion evoked excitation and responses to L-glutamate (1-2 mM) and kainate (10-20 microM). 3. In the guinea pig, salicylate (0.6-10 mM) suppressed the compound action potential (CAP) and increased N1 latency at low but not high sound intensities. 4. In the lateral line salicylate action may involve an antagonism of the hair-cell transmitter on the afferent nerve. 5. In the cochlea salicylate may suppress the active process or cochlear amplifier.

Comparative actions of gaba and acetylcholine on the Xenopus laevis lateral line

The effects of GABA, acetylcholine and carbachol on the spontaneous activity of afferent nerve fibers in the lateral line of Xenopus laevis are characterized. Atropine and bicuculline were also tested on drug- and water motion-evoked activity. GABA (0.019-1.25 mM) suppressed and both acetylcholine (1.25-80 microM) and carbachol (1.25-40 microM) increased spontaneous activity. These actions were blocked by bicuculline (100 microM) and atropine (4 microM) respectively. Atropine (20 microM) and bicuculline (100 microM) had no effect on water motion-evoked activity. The results characterize actions of GABA and acetylcholine not previously described and provide evidence that does not support the hypothesis that GABA or acetylcholine are the afferent transmitter.

An energy-dependent step in aminoglycoside ototoxicity: prevention of gentamicin ototoxicity during reduced endolymphatic potential

Guinea pigs received a bolus of gentamicin (10 mM for 5 min) by perilymphatic perfusion which normally led to an irreversible loss of the cochlear microphonic potential (CM). Various experimental conditions that reduced the endolymphatic potential (EP) were then superimposed on the gentamicin application. Reversible reductions in EP (and, concomitantly, in CM) were induced by asphyxia (3 min), intravenous furosemide (50 mg/kg), and perilymphatic perfusion of aminooxyacetic acid (10 mM). When the administration of gentamicin was initiated at the time of maximal EP reduction the usual irreversible gentamicin-induced decline of CM was prevented. The results indicate that a metabolic process is essential in the expression of gentamicin toxicity. The data are consistent with the inhibition of an energy-dependent transport of the aminoglycoside. Alternatively, the data are also compatible with the hypothesis that entry of gentamicin into hair cells is prevented by a reduction in their transmembrane electrical potential.

The use of kainic acid for studying the origins of scalp-recorded auditory brainstem responses in the guinea pig.

Kainic acid was injected into the medial nucleus of the trapezoid body (MNTB) of guinea pigs to evaluate its use in studying generator loci of the scalp-recorded auditory brain stem response (ABR). Sound-evoked near-field potentials from the MNTB and far-field ABRs were recorded before, during and up to 2 h after the injections. Two hours post-injection, small amounts of kainic acid (0.25 nmol in 0.1 microliters of Ringer solution) resulted in neuronal destruction which histologically appeared confined to the MNTB. Larger amounts (10 nmol in 1.0 microliters) produced more extensive lesions. Regardless of the dose of kainic acid, near-field activity evoked by contralateral ear stimulation was almost totally abolished and ABR wave III amplitude was reduced by as much as 60%. In future studies, the use of excitotoxic amino acids to produce lesions within complex nuclear subdivisions of the auditory pathway may yield valuable information as to the relative contributions that brainstem structures make to the various waves comprising the ABR and about the behavioral effects that axon sparing lesions produce.