Spatial representation of corticofugal input in the inferior colliculus: a multicontact silicon probe approach.

The inferior colliculus (IC) is a well-established target of descending projections from the auditory cortex (AC). However, our understanding of these pathways has been limited by an incomplete picture of their functional influence within the three-dimensional space of the IC. Our goal was to study the properties and spatial representation of corticofugal input in the IC of guinea pigs with a high degree of spatial resolution. We systematically mapped neural activity in the IC using two types of silicon substrate probes that allow for simultaneous recording at multiple neural sites. One probe provided a high resolution in the dorsal-ventral plane and the other provided spatial resolution in the medial-lateral plane. Electrical stimulation of the ipsilateral AC produced excitatory responses in the IC with thresholds usually below 5-10 microA. First spike latencies were predominantly in the 6-20 ms range, although latencies from 3-5 ms were also observed. Broadly distributed unimodal spike patterns with modal latencies greater than 30 ms were occasionally seen. The excitatory responses to cortical stimulation were mostly unimodal and occasionally bimodal with a wide range of spike distribution patterns and response durations. Excitation was often followed by suppression of spontaneous activity. Suppression of acoustic responses was observed even when there was little or no response to electrical stimulation, suggesting spatial-temporal integration. A few of the responding neurons showed purely inhibitory responses to electrical stimulation, suggesting that there are disynaptic routes of corticocollicular inhibition. Detailed spatial mapping revealed that the response patterns and their durations had a characteristic spatial distribution in the IC.

Effects of contralateral sound stimulation on unit activity of ventral cochlear nucleus neurons.

The cochlear nucleus (CN) commissural connection represents the first opportunity for convergence of binaural information in the auditory brainstem. All major neuron types in the ventral CN (VCN) are innervated by a diverse population of cells in the contralateral VCN. This study examined the effect of contralateral sound stimulation on the spontaneous rates (SRs) of neurons in the VCN. Unit activity was recorded with silicon-substrate multichannel probes which allowed recordings from up to 16 sites simultaneously. On average, 30% of units showed short-latency (often only 2 ms greater than the latencies of ipsilateral sound-evoked responses) inhibition of SR by wideband contralateral noise bursts. Fewer units (4.5%) were excited by contralateral noise at sound levels low enough to exclude excitation by acoustic crossover. Both regular and irregular units in the anterior VCN (AVCN) and posterior VCN (PVCN) were inhibited by contralateral sound. Decrements in SR followed a monotonic function with increases in contralateral sound level, except where responses could be attributed to acoustic crossover. Restricting the contralateral noise bandwidth resulted in a frequency-specific inhibition, dominated by frequencies at and below the ipsilateral BF of the unit, consistent with anatomical findings of the tonotopic organization of the CN commissural pathway. The latencies of these effects are compatible with mono, di and tri-synaptic connections reflecting CN commissural pathway effects.

Ambulatory management of burns.

Burns often happen unexpectedly and have the potential to cause death, lifelong disfigurement and dysfunction. A critical part of burn management is assessing the depth and extent of injury. Burns are now commonly classified as superficial, superficial partial thickness, deep partial thickness and full thickness. A systematic approach to burn care focuses on the six "Cs": clothing, cooling, cleaning, chemoprophylaxis, covering and comforting (i.e., pain relief). The American Burn Association has established criteria for determining which patients can be managed as outpatients and which require hospital admission or referral to a burn center. Follow-up care is important to assess patients for infection, healing and ability to provide proper wound care. Complications of burns include slow healing, scar formation and contracture. Early surgical referral can often help prevent or lessen scarring and contractures. Family physicians should be alert for psychologic problems related to long-term disability or disfigurement from burn injuries.

A multichannel neural probe for selective chemical delivery at the cellular level.

A bulk-micromachined multichannel silicon probe capable of selectively delivering chemicals at the cellular level as well as electrically recording from and stimulating neurons in vivo has been developed. The process buries multiple flow channels in the probe substrate, resulting in a hollow-core device. Microchannel formation requires only one mask in addition to those normally used for probe fabrication and is compatible with on-chip signal-processing circuitry. Flow in these microchannels has been studied theoretically and experimentally. For an effective channel diameter of 10 microns, a channel length of 4 mm, and water as the injected fluid, the flow velocity at 11 torr is about 1.3 mm/s, delivering 100 pl in 1 s. Intermixing of chemicals with the tissue fluid due to natural diffusion through the outlet orifice becomes significant for dwell times in excess of about 30 min, and a shutter is proposed for chronic use. The probe has been used for acute monitoring of the neural responses to various chemical stimuli in guinea pig superior and inferior colliculus.

Deafness-induced plasticity in the mature central auditory system.

Studies in rats and guinea pigs indicate that local changes in inhibitory transmitters may underlie deafness-induced plastic changes in electrophysiological responsiveness of cells of the mature central auditory system. Following 21 days of bilateral deafness there is an increase in evoked Fos-immunoreactive neurones in the central nucleus of the inferior colliculus (CIC) to contralateral cochlear electrical stimulation, compared with normal or 1-day deafened animals. Deafness is also associated with a dramatic reduction in the population of CIC neurones that respond with suppression of activity to electrical stimulation. Moreover, in vivo microdialysis reveals a marked decrease in gamma-aminobutyric acid (GABA) release from the CIC cells in deafened animals. The results may have general implications for the mediation of central nervous system plasticity induced by deafferentation of sensory input.

In vivo release of neuroactive amino acids from the inferior colliculus of the guinea pig using brain microdialysis.

Microdialysis techniques were used to measure in vivo release of neuroactive amino acids from the central nucleus of the inferior colliculus (ICC) in anesthetized guinea pigs. Concentric dialysis probes were implanted in the ICC and perfused with Ringer solution of various compositions at a flow rate of 2.0 microliters/min. Consecutive 10-min fractions of the dialysate were collected for up to 3 h under different experimental conditions, frozen and assayed for amino acid content by high performance liquid chromatography (HPLC). There was an initial high outflow of amino acids which declined to stable baseline levels after 2 h. Following this stabilization period, perfusion with a medium containing 100 mM KCl produced an increase in the extracellular levels of aspartate (Asp), glutamate (Glu), gamma-aminobutyric acid (GABA) and glycine (Gly). Only the increases in GABA and Gly were statistically significant. None of the increases occurred in the presence of 2.0 mM cobalt suggesting the release of amino acids is calcium dependent. Histological examination revealed that tissue damage was minimal and largely confined to the immediate vicinity of the probes. We were also able to show that the blood brain barrier (BBB) appeared to heal 2 h after probe implantation. Thus, following intravenous injection of [3H]alpha-aminoisobutyric acid (AIB), which does not cross the intact BBB, no isotope was recovered in the dialysate. These results demonstrate that microdialysis is a unique and suitable method to monitor changes in the extracellular levels of amino acid neurotransmitters in a central auditory structure.(ABSTRACT TRUNCATED AT 250 WORDS)

Connections between the cochlear nuclei in guinea pig.

This study provides a detailed analysis of the appearances and distributions of neurons projecting from one cochlear nucleus to the other. Injections of wheatgerm agglutinin conjugated to horseradish peroxidase were made into ventral or dorsal cochlear nucleus of the guinea pig. Retrogradely labeled cells in the opposite cochlear nucleus were examined and quantified. Three major categories of labeled cells were discerned on the basis of their soma shape: elongate, round-to-oval, and polygonal. All injections resulted in widespread labeling of cells in all of these categories, but especially round-to-oval cells, in the opposite ventral cochlear nucleus and sparse labeling in the dorsal cochlear nucleus. The results suggest that there is a significant cochlear nucleus commissural projection involving heterogeneous cell types which could have diverse functions in binaural auditory signal processing.

Patterns of glutamate, glycine, and GABA immunolabeling in four synaptic terminal classes in the lateral superior olive of the guinea pig.

The goal of this study was to correlate synaptic ultrastructure with transmitter specificity and function in the lateral superior olive (LSO), a nucleus that is thought to play a major role in sound localization. This was accomplished by means of postembedding immunogold immunocytochemistry. Four classes of synaptic terminals were identified in the LSO. They were distinguishable from one another both morphologically and on the basis of their different patterns of immunolabeling for glutamate, glycine, and gamma-aminobutyric acid (GABA). The highest level of glutamate immunoreactivity was found in terminals that contained round vesicles (R) and formed synaptic contacts with asymmetric synaptic junctions. Round-vesicle terminals predominated on small caliber dendrites by a ratio of at least 2:1 over the other classes combined. The thinnest dendrites were typically contacted by R terminals only. The ratio of R terminals to the other types decreased as the caliber of the dendritic profiles they apposed increased so that on the soma, R terminals were outnumbered by at least 2:1 by the other types. Terminals containing flattened vesicles (F) exhibited intense immunoreactivity for both glycine and glutamate, although the glutamate immunolabeling was not as high as that in the R terminals. Flattened-vesicle terminals formed symmetric synaptic contacts with their targets and their distribution was the reverse of that described for R terminals; i.e., they were most abundant on LSO perikarya and fewest on small caliber dendrites. Two terminal types, both containing pleomorphic vesicles and forming symmetric synaptic junctions, were found in far fewer numbers. One group contained large pleomorphic vesicles (LP) and was immunoreactive for both glycine and GABA. The other group contained small pleomorphic vesicles (SP) along with a few dense-core vesicles and labeled for GABA only. The LP terminals were preferentially distributed on somata and large-caliber dendrites, while the SP terminals most often contacted smaller dendrites. Previous work suggests that a large percentage of the R terminals arise from spherical cells in the ipsilateral cochlear nucleus and are excitatory in action. This pathway may use glutamate as a transmitter. Many of the F terminals are thought to originate from the ipsilateral medial nucleus of the trapezoid body and appear to be the inhibitory (glycinergic) terminals from a pathway that originates from the contralateral ear. The origins and functions of LP and SP terminals are unknown, but a few possibilities are discussed along with the significance of cocontainment of neuroactive substances in specific terminal types.

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.

Acetylcholine, carbachol, and GABA induce no detectable change in the length of isolated outer hair cells.

The mechanical and electrical properties of cochlear outer hair cells (OHCs) are suggested to modulate transduction by inner hair cells. These properties of OHCs are presumably regulated by efferent neurons which use several transmitters including acetylcholine (Ach) and gamma aminobutyric acid (GABA). Since it had been suggested that Ach causes isolated OHCs to shorten visibly, this study was designed to investigate whether GABA also alters the length of OHCs. OHCs were isolated from the guinea pig cochlea by mechanical dispersion after collagenase treatment. Cells were initially selected by strict morphological criteria. In addition they were only included in further studies if they attained a constant length during 10 min of superfusion with buffer solution. Neither GABA (20 microM: 100 microM), Ach (5 mM; 10 microM with 10 microM eserine) or carbachol (10 microM; 100 microM) altered OHC length when applied in iso-osmotic Hank's balanced salt solution (total number of cells tested, 72). If a change in length occurred it must have been smaller than 0.3 microns, our detection ability. In contrast, high potassium and variations in osmolarity changed hair cell length by 3-10% in agreement with other reports.

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.

Potassium-induced release of endogenous glutamate and two as yet unidentified substances from the lateral line of Xenopus laevis.

The release of endogenous glutamate and other primary amines from the lateral-line of Xenopus laevis was studied using an in vitro superfusion technique and high performance liquid chromatography. Potassium stimulation (50 mM KCl) applied to 60 or 120 lateral-line organs dissected from the skin and pooled in a perfusion chamber induced the release of glutamate and aspartate. The release of aspartate was smaller than that of glutamate and more variable. A variable release of two, as yet, unidentified substances was also detected. In low calcium (0.1 mM CaCl2), high magnesium (10 mM MgCl2) solution, 50 mM potassium failed to induce an increase in glutamate, aspartate and the two unknowns, suggesting they are released in a transmitter-like manner. The technique presents a new and simple method for studying transmitters in hair-cell systems. Although other interpretations are possible, the results are consistent with the hypothesis that glutamate is a hair-cell transmitter and suggest a potential role for other substances in the transduction process, perhaps as neuromodulators.

Analysis of histamine as a hair-cell transmitter in the lateral line of Xenopus laevis.

The actions of histamine and histamine antagonists on afferent nerve activity were investigated in the lateral line of Xenopus laevis. Histamine (0.002-2.0 mM) had no effect on spontaneous activity or excitatory responses to water motion. In contrast, pyrilamine, an H1 receptor antagonist, suppressed spontaneous activity beginning at 0.01-0.05 mM. Below 0.3 mM the suppression was often preceded by a small excitatory response and responses to high (24-30 dB re threshold), but not low (0-18 dB) levels of water motion were selectively suppressed. Higher concentrations (0.3-2.0 mM) abolished spontaneous activity and suppressed responses at all levels of water motion. Cimetidine, an H2 receptor antagonist, had similar actions but was one-tenth as potent as pyrilamine. Tetrodotoxin (0.001-0.1 microM), which blocks voltage-sensitive Na+ channels, mimicked the suppressive effects of the histamine antagonists. Histamine (2.0 mM) failed to block the actions of pyrilamine (0.1 mM) indicating its effects are mediated through a mechanism other than histamine receptors. In addition, pyrilamine (0.05-0.1 mM) non-selectively suppressed excitation to exogenously applied L-glutamate (1.0-2.0 mM), L-aspartate (1.0-2.0 mM), kainate (0.005-0.01 mM), and quisqualate (0.002-0.005 mM) and altered responses to N-methyl-D-aspartate (0.5-1.0 mM). The results are inconsistent with histamine being a transmitter in the Xenopus lateral line and reveal that the actions of histamine antagonists are nonspecific, possibly due, in part, to blockade of voltage-sensitive Na+ channels.

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.

Intraoperative facial nerve monitoring: a comparison of stimulating electrodes.

Preservation of the facial nerve during acoustic neuroma resection may be enhanced by the use of intraoperative electrical stimulation. Although stimulation of the extratemporal facial nerve is an effective and established procedure, anatomic differences of the intradural facial nerve and its microenvironment demand more exacting stimulus protocols. The absence of epineurium may make the intradural nerve more susceptible to mechanical or electrical trauma while intermittent pooling of cerebrospinal fluid (CSF) at the cerebellopontine angle may shunt current away from nerve. Four stimulus configurations were examined under varying conditions simulating CSF pooling. The results indicated that: 1. insulation of stimulating electrodes prevents CSF current shunting and allows utilization of a constant current source, and 2. monopolar and bipolar configurations demonstrate significantly different electrical characteristics which may be employed selectively based upon specific clinical goals.

Antidromically evoked facial nerve response.

Facial nerve electrodiagnostic tests that are currently available indirectly assess the severity of injury to the intratemporal facial nerve. Antidromic conduction testing is an alternate approach that, if feasible, could provide direct and immediate assessment of proximal facial nerve function. This possibility was tested in a guinea pig model, in which near-field (intracranial) and far-field (extradural) recording techniques were used to assess antidromically evoked facial nerve activity. Response characteristics, topographical distribution, and lesion effects suggest that the recorded potentials represent antidromic activation of the facial nerve. If response amplitude and/or latency can be correlated with the functional state of the nerve, antidromic testing may provide a useful means of assessing proximal facial nerve function in pathologic states.

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.

Comparative actions of glutamate and related substances on the lateral line of xenopus laevis.

The effects of compounds structurally related to L-glutamate were compared on spontaneous activity of afferent nerve fibres in Xenopus laevis lateral-line. The potencies (EPMR) of several compounds relative to L-glutamate (EPMR = 1) were: L-aspartate (EPMR = 1), D-aspartate (EPMR = 1), D-glutamate (EPMR = 1.98), quisqualate (EPMR = 0.0015), kainate (EPMR = 0.0045), dihydrokainate (EPMR = 0), ibotenate (EPMR = 1.5), L-homocysteate (EPMR = 0.17), 2,4,5-trihydroxyphenylalanine (6-OH-DOPA, EPMR = 0.23), and D-homocysteate (EPMR = 0.73), N-methyl-D,L-aspartate (EPMR = 5), and N-methyl-D-aspartate (NMDA, EPMR = 1). Several compounds, including gamma-aminobutyrate (GABA), suppressed spontaneous activity without eliciting excitatory responses. The results reveal receptors that are present at this vertebrate peripheral synapse.

Effects of D-alpha-aminoadipate on excitation of afferent fibers in the lateral line of Xenopus laevis.

The effects of D-alpha-aminoadipate (D alpha AA) on excitation of afferent nerve fibers in the Xenopus laevis lateral line were studied in vitro. D alpha AA reversibly suppressed spontaneous activity and excitation induced by water motion at concentrations as low as 0.25-0.5 mM. Higher concentrations (up to 10 mM) caused a greater suppression that was rapidly and fully reversible. L-alpha-Aminoadipate at 0.25-1.0 mM had no suppressive effects. Responses elicited by NMDA (1.0-2.0 mM) were the most sensitive to D alpha AA (0.25-0.5 mM), those elicited by L-aspartate and L-glutamate (1.0-2.0 mM) were less sensitive and similar, and those elicited by kainate (5-15 muM) were the least sensitive. The results provide evidence that the transmitter released by hair cells in the Xenopus lateral line interacts postsynaptically with NMDA-preferring receptors and that the transmitter is an excitatory amino acid, possibly L-glutamate or L-aspartate.