Immunohistochemical study on the distributions of AMPA receptor subtypes in rat vestibular nuclear complex after unilateral deafferentation.
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Afferent Pathways↗
Biomedical subjects
D A Godfrey
Publications and source records attributed to D A Godfrey.
Quantitative changes of amino acid distributions in the rat vestibular nuclear complex after unilateral vestibular ganglionectomy.
Changes of amino acid concentrations in the vestibular nuclear complex (VNC) during lesion-induced vestibular compensation were studied in rats after unilateral vestibular ganglionectomy. Distributions of 12 amino acids within the VNC were measured at 2,4,7, and 30 days after surgery, using microdissection of freeze-dried brain sections and HPLC. Glutamate decreased on the lesioned side in nearly all VNC regions. Changes were fully developed 2 days after lesion and persisted through 30 days. In some regions, glutamate decreased also on the unlesioned side, especially at longer survival times, so that bilateral asymmetries became reduced. Aspartate changes were similar to those of glutamate on either side. Lesion-induced glutamine asymmetry was usually opposite to that of glutamate. Although GABA concentration decreased at early survival times, it recovered at later times and sometimes increased in dorsal parts of lateral and medial nuclei. Taurine changes were similar to those of GABA in most regions. Glycine change was primarily limited to a bilateral decrease in the dorsal part of the lateral vestibular nucleus. Concentrations of other amino acids were much lower, but some showed postlesion changes.
Amino Acids↗
Amino acid concentrations and selected enzyme activities in rat auditory, olfactory, and visual systems.
Homogenates of specific brain regions of three sensory systems (auditory, olfactory, and visual) were prepared from pigmented Long-Evans Hooded rats and assayed for amino acid concentrations and activities of glutaminase, aspartate aminotransferase (total, cytosolic, and by difference, mitochondrial), malate dehydrogenase, lactate dehydrogenase, and choline acetyltransferase. Comparing the quantitative distributions among regions revealed significant correlations between AAT and aspartate, between glutaminase and glutamate, between glutamate and glutamine, and between AAT plus glutaminase, or glutaminase alone, and the sum of aspartate, glutamate, and GABA, suggesting a metabolic pathway involving the synthesis of a glutamate pool as precursor to aspartate and GABA. Of the inhibitory transmitter amino acids, GABA concentrations routinely exceeded those of glycine, but glycine concentrations were relatively high in brainstem auditory structures.
Amino Acids↗
Immunohistochemistry of muscarinic acetylcholine receptors in rat cochlear nucleus.
Neurons of the cochlear nucleus (CN) receive extrinsic and intrinsic cholinergic inputs, the effects of which appear to be mediated primarily by muscarinic acetylcholine receptors (mAChRs). To investigate the distribution of mAChRs and the correlation of pre- and post-synaptic cholinergic markers in CN, we used a monoclonal anti-mAChR antibody (M35) and a monoclonal antibody against choline acetyltransferase (ChAT) to perform immunohistochemistry on rat brain sections, and we also carried out histochemistry for acetylcholinesterase (AChE) activity. The density distributions of ChAT immunohistochemistry and AChE activity histochemistry agreed well with previous quantitative measurements of the activity distributions of these enzymes. A generally close correlation between densities of M35 and ChAT immunoreactivities was found across subregions of rat CN. The localizations of M35 and ChAT immunoreactivities suggest that cholinergic synapses are mostly axosomatic in ventral CN and possibly mostly axodendritic in dorsal CN. Prominent differences in density and appearance between M35 immunoreactivity and AChE activity were found in the regions of the rat CN where granule cells predominate.
Acetylcholinesterase↗
Muscarinic receptor subtypes in rat dorsal cochlear nucleus.
We previously reported that responses of spontaneously active rat dorsal cochlear nucleus (DCN) neurons to cholinergic agonists are mediated predominantly by muscarinic receptors. We have now tested the effects of 7 antagonists with differing affinities for the muscarinic receptor subtypes M1-M4 on the responses to constant, submaximal doses of carbachol in rat brainstem slices. Each slice was exposed to one or more concentrations of one antagonist applied during extracellular recording of a DCN neuron. The concentrations yielding 50% reduction of test responses (IC50) of regular and bursting neurons were estimated for each antagonist. Correlation coefficients were calculated between log(IC50) values and log(Ki) values of the drugs for the receptor subtypes. Correlation coefficients for both regular and bursting neurons were not significant (P > 0.05) for M1 and M3, but were significant (P < 0.02) for M4. Bursting but not regular neurons also showed a significant correlation for M2 (P < 0.05). Our results suggest that (1) M4 contributes to the cholinergic responses in DCN and M2 may also contribute to the responses of bursting neurons, but the contribution of other subtypes cannot be completely excluded; (2) muscarinic subtypes in DCN probably differ from those reported for cochlea and some brain regions.
Animals↗
Comparison of surgeries for removal of primary vestibular inputs: a combined anatomical and behavioral study in rats.
Unilateral removal of Scarpa's ganglion and neurectomy of the peripheral vestibular nerve branches were compared in rats as methods to eliminate primary vestibular input. Ocular nystagmus was consistently observed after both types of lesion, but it completely disappeared within 4 to 7 days. Imbalance and rotation were more serious and prolonged after ganglionectomy than after peripheral neurectomy. Corresponding with these differences in symptoms were differences in terminal degeneration. After ganglionectomy, degenerated axons and terminals were distributed throughout all terminal regions of primary vestibular fibers on the lesioned side, while after peripheral neurectomy, the degeneration was more limited. The results of this study suggest that vestibular ganglionectomy is a more successful approach than peripheral vestibular neurectomy for removing the primary vestibular input.
Afferent Pathways↗
Cholinergic modulation of spontaneous activity in rat dorsal cochlear nucleus.
Extracellular recordings were made from brain stem slices to test the effects of bath application of cholinergic agonists and antagonists on the firing rates of spontaneously active dorsal cochlear nucleus neurons. About 90% of neurons responded to carbachol. A higher proportion responded to muscarine than to nicotine. Muscarine elicited larger responses at lower concentrations than nicotine. Responses to either carbachol or muscarine were always blocked by atropine or scopolamine. The nicotinic antagonists d-tubocurarine, hexamethonium, and mecamylamine blocked the responses to nicotine, but did not decrease the responses to carbachol. Regularly firing neurons showed only increases of firing rate during exposure to cholinergic agonists. About half of responsive bursting neurons showed increased firing; half showed increased followed by decreased firing to 10 microM carbachol or muscarine. All phases of the responses of most bursting neurons were greatly decreased or abolished in low calcium, high magnesium medium, while responses of regular neurons were not detectably affected. Thus, cholinergic agonists appear to act directly on regularly firing neurons, while their actions on bursting neurons may require synaptic activity. The data suggest that cholinergic transmission in the dorsal cochlear nucleus is predominantly muscarinic, and that most regularly firing spontaneously active neurons have muscarinic receptors.
Action Potentials↗
Aspartate aminotransferase and glutaminase activities in rat olfactory bulb and cochlear nucleus; comparisons with retina and with concentrations of substrate and product amino acids.
The quantitative distributions of aspartate aminotransferase and glutaminase were mapped in subregions of olfactory bulb and cochlear nucleus of rat, and were compared with similar data for retina and with the distributions of their substrate and product amino acids aspartate, glutamate, and glutamine. The distributions of both enzymes paralleled that of aspartate in the olfactory bulb and that of glutamate in the cochlear nucleus. In retina (excluding inner segments), there were similarities between aspartate aminotransferase and both glutamate and aspartate distributions. The distribution of gamma-aminobutyrate (GABA) was similar to those of both enzymes in olfactory bulb, to aspartate aminotransferase in cochlear nucleus, and to glutaminase in retina (excluding inner segments). The results are consistent with significant involvement of aspartate aminotransferase, especially the cytosolic isoenzyme, and glutaminase in accumulation of the neurotransmitter amino acids glutamate, aspartate, and GABA, although with preferential accumulation of different amino acids in different brain regions.
Amino Acids↗
Functional characteristics of spontaneously active neurons in rat dorsal cochlear nucleus in vitro.
1. The cochlear nucleus of rat brain stem slices was explored with extracellular microelectrodes to determine the distribution and characteristics of spontaneously active neurons. 2. In mapping experiments few spontaneously active neurons were found in anteroventral or posteroventral divisions of the cochlear nucleus. In contrast, spontaneously active neurons (N = 648) were widely distributed in the dorsal cochlear nucleus (DCN), especially its more superficial part. The density (neurons per penetration) was greatest 100-400 microns from the lateral surface of DCN, corresponding approximately to the fusiform soma layer and closely adjacent portions of the molecular and deeper regions. In penetrations with active neurons as many as 13 were found, with a mean of 4.3 neurons per penetration. Activity was found along the entire dorsomedial-ventrolateral extent of the nucleus, across the tonotopic representation. 3. Most neurons were readily categorized according to the spike interval pattern as regular (40%), bursting (30%), or irregular (30%). Regular and bursting patterns were highly stable, but few bursting neurons were found in relatively inactive slices. Although there was extensive overlap in location, bursting neurons were significantly closer to the lateral edge of the slice. Also, they were more likely to have initially negative action potentials than regular or irregular neurons. 4. A high density of spontaneous firing, including regular, bursting, and irregular patterns, was observed in slices containing only DCN and adjacent fiber tracts, with other nuclear structures trimmed away. 5. When the K+ concentration of the perfusion medium was decreased from 6.25 to 3.25 mM firing rates of regular neurons decreased moderately without changes in pattern. In contrast, firing rates of most bursting and irregular neurons showed large increases, and bursts were prolonged. 6. When the K+ concentration was increased from 6.25 to 9.25 or 12.25 mM regular neurons showed moderate increases in rate without changes in pattern. Effects on firing rates differed among bursting and irregular neurons, but bursts usually increased in frequency and decreased in duration, and irregular neurons showed some burst firing. 7. When Ca2+ was decreased to 0.2 mM and Mg2+ increased to 3.8 or 7.8 mM regular neurons did not change in pattern of firing although firing rates increased or decreased moderately. Bursting neurons showed large increases in the durations of the bursts. Firing rates of bursting neurons usually increased during 0.2 mM Ca2+ -3.8 mM Mg2+ but typically decreased, after an initial rise, during 0.2 mM Ca2+ -7.8 mM Mg2+.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Choline acetyltransferase in cochlear root neurons.
Cochlear root neurons are a distinct group of cells located in the auditory nerve root in small rodents. Their transmitter is still unknown. Some of our preparations showed immunoreactivity of somata of cochlear root neurons with both polyclonal and monoclonal antibodies against choline acetyltransferase (ChAT) which, despite their very weak histochemical reaction for acetylcholinesterase (AChE), suggested that cochlear root neurons might be cholinergic. To test this, we used a radiometric assay to measure ChAT activities of rat auditory nerve root samples containing cochlear root neurons and of adjacent samples not containing them. There was no significant difference between the low mean ChAT activities of these two groups of samples. Thus, cochlear root neurons are not likely to be cholinergic.
Animals↗
Effect of copper-deficient diet on metabolism in rat auditory structures.
Copper is a trace element known to be critical for normal brain function, and abnormal copper metabolism has been associated with some disorders involving the auditory system. We examined effects of copper deficiency on metabolism in major structures of the auditory system. Homogenates of cochlea, cochlear nucleus and inferior colliculus of rats, as well as whole brain, were assayed for activities of enzymes of oxidative and glycolytic energy metabolism--malate and lactate dehydrogenase, enzymes of acetylcholine metabolism--choline acetyltransferase and acetylcholinesterase, and concentrations of amino acids. Whole brain was also assayed for activity of superoxide dismutase, a copper-containing enzyme, and concentrations of minerals. For these chemicals and tissues, the only significant differences between copper-deficient and copper-adequate rats were: (1) decreased copper and magnesium and increased potassium concentrations in whole brain of copper-deficient rats and (2) an elevation of glutamine concentration in inferior colliculus and whole brain of copper-deficient rats. The elevated glutamine could not be related to any change in activity of glutamine synthetase or glutaminase, major enzymes of glutamine metabolism. It is speculated that the increase in glutamine might result from a net increase in ammonia accumulation in the brains of copper-deficient rats.
Amino Acids↗
Sampling fluid from slice chambers by microsiphoning.
We have developed a simple technique that continuously samples perfusion fluid from an in vitro slice chamber and permits simultaneous electrophysiological recordings from a slice. This procedure uses a microsiphon that diverts approximately 1.5% of the total effluent of artificial cerebrospinal fluid from an interface slice chamber into a collection tube at a lower level outside the chamber. The tip of the microsiphon may be moved anywhere within the chamber without disturbing the slice. For optimum sampling of the release of substances from one area of a slice, the tip is positioned immediately downstream from the location being studied. Concentrations of several amino acids were measured to identify significant spatial and temporal characteristics of the technique and to demonstrate that spontaneous and induced release of amino acids from slices can be measured with adequate sensitivity and time resolution.
Amino Acids↗
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.
Animals↗
Distributions of choline acetyltransferase and acetylcholinesterase activities in the retinal layers of the red-tailed hawk and road runner.
The activities of choline acetyltransferase and acetylcholinesterase were assayed in submicrogram samples from layers of red-tailed hawk and road runner retina. Both enzyme activities were concentrated in and near the inner plexiform layer. Within the inner plexiform layers of both species, activities of each enzyme were concentrated in two bands, one in each half of this layer. Little choline acetyltransferase activity was found superficial to the middle third of the inner nuclear layer. The distributions of acetylcholinesterase activities corresponded well to those of choline acetyltransferase, except in the outer plexiform layer and the outer margin of the inner nuclear layer of the hawk. These distributions of enzyme activities indicate that populations of amacrine cells in the retinae of these species are cholinergic. In addition to these same cells and presumably cholinoceptive amacrine and ganglion cells, acetylcholinesterase activity in the hawk was associated with a population of horizontal cells that may be unrelated to synaptic cholinergic neurotransmission. Choline acetyltransferase activities associated with amacrine somata and processes were about four times greater in the hawk than in the road runner, suggesting important differences in the density and function of cholinergic elements between species. Possible synaptic relationships in the inner plexiform layer consistent with the interspecies differences in enzyme activities are considered.
Acetylcholinesterase↗
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.
Animals↗
Distributions of choline acetyltransferase and acetylcholinesterase activities in the retinal layers of pigeon red and yellow fields.
The activities of choline acetyltransferase and acetylcholinesterase were assayed in submicrogram samples from layers of pigeon retina. Red and yellow fields were sampled separately to investigate quantitatively the relationship between these enzymes of acetylcholine metabolism and the gradient of inner plexiform layer complexity, increasing from the yellow field to the red. Choline acetyltransferase and acetylcholinesterase activities were concentrated in and near the inner plexiform layer, within which two peaks of activity for each enzyme were obtained. The distributions of enzyme activities indicate that populations of amacrine cells in the pigeon retina are cholinergic. The quantitative similarities between the enzyme activities in red and yellow fields suggest that the cholinergic system may not be specifically involved in the increase in inner plexiform layer complexity across the pigeon retina.
Acetylcholinesterase↗
Enzymes of transmitter and energy metabolism in rat middle ear and extraocular muscles.
To further investigate the peculiar characteristics of the middle ear and extraocular muscles, compared to the extensively studied skeletal muscles of the limbs, activities of enzymes of transmitter and energy metabolism were measured in homogenates of these muscles from albino and pigmented rats. These activities were compared to those for a masticatory muscle and for three hindlimb muscles chosen for their preponderance of either slow oxidative, fast glycolytic, or fast oxidative glycolytic fibers. Activities of the neuromuscular transmitter enzymes choline acetyltransferase and acetylcholinesterase were relatively very high in the extraocular and middle ear muscles. The activity of malate dehydrogenase, an enzyme of oxidative energy metabolism, was as high in the extraocular, masticatory and stapedius muscles as in the oxidative hindlimb muscles, but was lower in tensor tympani. The activity of lactate dehydrogenase, an enzyme of glycolytic energy metabolism, was remarkably low in both middle ear muscles. The results are consistent with high innervation density in the extraocular and middle ear muscles, and highly oxidative metabolism in the extraocular and stapedius muscles. Metabolic differences between the stapedius and tensor tympani suggest a relatively more active role for the former in the function of the rat middle ear.
Animals↗
Contribution of centrifugal innervation to choline acetyltransferase activity in the cat cochlear nucleus.
Using a quantitative microchemical mapping approach combined with surgical cuts of fiber tracts, the contributions of centrifugal pathways to choline acetyltransferase activity were mapped three-dimensionally in the cat cochlear nucleus. Large reductions of choline acetyltransferase activity, averaging 70%, were measured in almost all parts of the lesion-side nucleus following transection of virtually all its centrifugal connections. More superficial cuts, penetrating just through the olivocochlear bundle, also led to significant reductions of enzyme activity, especially most rostrally in the anteroventral cochlear nucleus and superficial granular region, where the reductions were similar to those following the complete cuts. Lesions encroaching upon the superior olivary complex gave bilateral effects. Transverse cuts between rostral and caudal parts of the cochlear nucleus gave some small effects. The results suggest that, as in rats, most choline acetyltransferase activity in the cat cochlear nucleus is associated with its centrifugal innervation. However, unlike the situation in rats, the enzyme activity in cats is related more to olivocochlear branches than to ventral fibers in the trapezoid body region. Also, the choline acetyltransferase activity related to olivocochlear collateral innervation is much less uniformly distributed within the cochlear nucleus in cats than in rats.
Animals↗