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A J Beitz

Publications and source records attributed to A J Beitz.

At least 91 records · Page 5Linked to original sources

A quantitative light microscopic analysis and ultrastructural description of cholecystokinin-like immunoreactivity in the spinal trigeminal nucleus of the rat.

The spinal trigeminal nucleus is involved in orofacial sensory transmission. Cholecystokinin octapeptide has been identified in axons in this nucleus and appears to play a role in the transmission of orofacial sensation from the trigeminal ganglia to the spinal trigeminal nucleus. Although cholecystokinin has been reported in axonal processes within the spinal trigeminal nucleus at the light microscopic level, nothing is known about the synaptic relationships of these cholecystokinin axons. The goals of this study were to quantitatively determine the volume fraction of cholecystokinin-like immunoreactive cell bodies and fibers in the three subnuclei of the spinal trigeminal nucleus, to provide the first ultrastructural description of cholecystokinin-like immunoreactive processes within these subnuclei and to analyse the synaptic relationships of cholecystokinin-like immunoreactive processes within the spinal trigeminal nucleus neuropil. Cholecystokinin-like immunoreactivity was localized by the peroxidase-antiperoxidase method or the peroxidase labeled, avidin-biotin technique and quantified at the light microscopic level by point counting. Immunoreactive fibers were present in all three subnuclei, but the greatest volume fraction of immunoreactive axons was obtained in laminae I and II of the nucleus caudalis. No immunoreactive cell bodies were evident in any of the subnuclei. The majority of immunoreactive profiles in all three subnuclei were identified ultrastructurally as axon terminals that contained both small and medium sized agranular vesicles and infrequently, large dense core vesicles. These immunoreactive terminals were usually found in close contact with non-immunoreactive dendrites with which they were observed to form asymmetric synapses. Immunoreactive terminals were occasionally observed to contact the cell bodies of large non-immunoreactive neurons on the border of laminae I and II in the nucleus caudalis. These results indicate that cholecystokinin-like immunoreactive processes are present throughout the spinal trigeminal nucleus, and in nucleus caudalis show a distribution similar to that reported for the spinal cord dorsal horn. Immunoreactive axons make synaptic contact with both the dendrites and perikarya of spinal trigeminal nucleus neurons. No axoaxonic synapses were observed. These findings suggest that cholecystokinin plays an important role in spinal trigeminal nucleus function. The possible colocalization of cholecystokinin and substance P in the spinal trigeminal nucleus, and the possible role of cholecystokinin in attenuating the action of opioids in the spinal trigeminal nucleus are also discussed.

Animals↗

Localization of glutamate in trigeminothalamic projection neurons: a combined retrograde transport-immunohistochemical study.

Trigeminothalamic projection neurons are important components of the pathways for conscious perception of pain, temperature, and tactile sensation from the orofacial region. The neurotransmitters utilized by trigeminal neurons projecting to the thalamus are unknown. By use of a monoclonal antibody specific for fixative-modified glutamate and a polyclonal antiserum against glutaminase, we recently identified neurons in the trigeminal sensory complex that contain glutamate-like immunoreactivity (Glu-LI) and glutaminase-like immunoreactivity. In the present study, we utilized combined retrograde transport-immunohistochemical techniques to localize putative glutamatergic trigeminothalamic neurons. Following injection of the retrograde tracer, wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP), into the ventroposterior medial thalamus (VPM), the number of neuronal profiles that were double-labeled with WGA:HRP and Glu-LI was greatest in principal sensory nucleus (Pr5), followed by subnuclei interpolaris (Sp5I) and caudalis (Sp5C). The average percentages of projection neurons double-labeled with Glu-LI were approximately 60-70% in Pr5 and Sp5I and 40% in Sp5C. The majority of double-labeled profiles in Sp5C were located in the magnocellular layer, as opposed to the marginal and substantia gelatinosa layers. A large injection site that spread into the intralaminar thalamic nuclei and nucleus submedius--areas implicated in the processing of nociceptive information--resulted in an increase in the ratio of single-labeled to double-labeled projection profiles in Sp5C. These results suggest that glutamate may be the neurotransmitter for a majority of trigeminothalamic projection neurons located in Sp5I and Pr5. However, on the basis of anatomical association, glutamate does not appear to be the major transmitter for neurons in Sp5C that forward nociceptive information to the thalamus.

Afferent Pathways↗

Monoclonal antibodies specific for fixative-modified aspartate: immunocytochemical localization in the rat CNS.

Aspartate is a putative excitatory amino acid neurotransmitter that is widely distributed in the CNS. To study its distribution, monoclonal antibodies were produced against beta-L-aspartyl-L-aspartate (beta-Asp-Asp) conjugated to keyhole limpet hemocyanin (KLH) using glutaraldehyde-borohydride. Three monoclonal antibodies, Asp1-3, were obtained with high degrees of selectivity for aldehyde-fixed aspartate. The immunocytochemical staining pattern of rat CNS was found to be similar for all 3 antibodies but differed in some regions from staining patterns produced by Glu1, a monoclonal antibody with high selectivity for a form of amide-linked glutamate. Tissue staining produced by Asp1-3 could be inhibited using aspartate conjugated to carrier proteins. Staining by Asp1 and Asp2 was also inhibited by free small molecules containing aspartate. Specificity of the 3 antibodies was evaluated by enzyme-linked immunoassay (ELISA) as follows: (1) reactivity of antibodies for conjugates of small molecules coated on ELISA plates; (2) ability of free small molecules to inhibit reactivity of antibodies for beta-Asp-Asp/KLH coated on ELISA plates; and (3) ability of conjugates to inhibit reactivity of antibodies for beta-Asp-Asp/KLH coated on ELISA plates. In all 3 types of assays, Asp1 and Asp2 displayed strong reactivity for small molecules and conjugates containing aspartate and little reactivity for small molecules and conjugates containing glutamate or GABA. Asp3 was highly reactive with conjugates containing aspartate using both directed and inhibition ELISA assays. For all 3 antibodies the precise staining pattern varied with the fixative used. Following glutaraldehyde fixation, dense immunocytochemical staining was observed in cerebral cortical neurons, some cerebellar granule cells, hippocampal pyramidal cells, and neurons of the inferior olivary nucleus. In addition, some putative GABAergic neurons, e.g., cerebellar basket and stellate cells, appeared to be stained. In general, acrolein fixation resulted in a more selective staining pattern in the CNS. For example, putative GABAergic neurons were no longer immunoreactive nor were hippocampal pyramidal cells.

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Differential origin of brainstem serotoninergic projections to the midbrain periaqueductal gray and superior colliculus of the rat.

Previous studies have shown that both the midbrain periaqueductal gray (PAG) and the superior colliculus receive a significant serotoninergic (5-HT) innervation. In the present study the origins of these 5-HT projections to the rodent PAG and superior colliculus were analyzed by using a combined immunohistochemical-retrograde transport technique. Thirteen brainstem regions were found to contain double-labelled 5-HT-like immunoreactive neurons following HRP injections into the PAG while only four brainstem nuclei contained double-labelled neurons following superior collicular injections. After HRP deposits into the ventral PAG, the largest percentage of double-labelled neurons was identified in nucleus raphe magnus, pars alpha of the nucleus gigantocellularis, and the paragigantocellular nucleus. The dorsal PAG, on the other hand, received the largest percentage of its 5-HT projections from nuclei raphe dorsalis, raphe obscurus, raphe pontis, and raphe medianis. The 5-HT input to the superior colliculus was found to arise exclusively from nuclei raphe dorsalis, raphe medianis, and raphe pontis and from the contralateral periaqueductal gray. Raphe nuclei were found to contribute serotoninergic projections to both the PAG and the superior colliculus while reticular nuclei contributed 5-HT projections only to the PAG. Injections of the fluorescent retrograde tracers true blue and nuclear yellow were then made into the PAG and superior colliculus to ascertain if neurons located in raphe nuclei that projected to both structures provided axon collaterals to both areas. Generally, less than 10% of raphe neurons projecting to the superior colliculus were identified as providing axon collaterals to the PAG. The present results demonstrate major quantitative and qualitative differences in the origin of 5-HT projections to the ventral PAG and superior colliculus. The origin of 5-HT input to the dorsal PAG, on the other hand, showed many similarities to the origin of 5-HT innervation of the superior colliculus. These data also indicate that approximately 35% of raphe neurons provide nonserotoninergic projections to the PAG and superior colliculus.

Afferent Pathways↗

Co-localization of fixative-modified glutamate and glutaminase in neurons of the spinal trigeminal nucleus of the rat: an immunohistochemical and immunoradiochemical analysis.

The spinal trigeminal nucleus (STN) is involved in processing orofacial sensory information, including tactile, thermal and nociceptive input, and relaying this information to higher brain centers, such as the thalamus. Very little information is available regarding the major excitatory neurotransmitters of this nucleus. The amino acid glutamate has been proposed as a major excitatory neurotransmitter in the central nervous system. In the present study, a novel monoclonal antibody, specific for fixative-modified glutamate, was utilized in conjunction with polyclonal antisera against glutaminase and aspartate aminotransferase (AATase) in an attempt to identify and map the locations of possible glutamatergic neurons in the STN. Co-localization experiments were performed by radiolabeling our monoclonal antibody and using this antibody in conjunction with the polyclonal antisera against glutaminase and AATase to evaluate the possible coexistence of glutamate with glutaminase or AATase in STN neurons. In all three subnuclei of the STN, immunohistochemically labeled neuronal profiles were observed with both of the polyclonal antisera and with the monoclonal antibody. Subnucleus caudalis contained the greatest number of labeled profiles per coronal section followed by subnucleus interpolaris and subnucleus oralis. The number and the distribution of immunoreactive profiles observed after the use of the glutaminase antiserum was comparable to that obtained with the monoclonal antibody. Co-localization experiments demonstrated that all glutaminase-like immunoreactive neurons also contained fixative-modified glutamate-like immunoradioactivity. These results suggest that glutamatergic neurons are present in the spinal trigeminal nucleus. The AATase antiserum labeled more neuronal profiles in each of the three subnuclei than did the glutaminase antiserum or the monoclonal antibody. In addition, co-localization experiments indicated that glutamate-like immunoreactivity was present in only two-thirds of AATase-like immunoreactive neuronal profiles. These findings suggest that glutaminase may be a more reliable marker of glutamatergic function than AATase.

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Immunocytochemical localization of glutamate-, glutaminase- and aspartate aminotransferase-like immunoreactivity in the rat deep cerebellar nuclei.

Although the anatomy and the connectivity of the deep cerebellar nuclei have been well documented, little is known about the neurotransmitter systems mediating cerebellar efferent pathways. The present study utilizes immunohistochemical procedures in conjunction with a novel monoclonal antibody specific for carbodiimide-fixed glutamate and polyclonal antisera against glutaminase (GLNase) and aspartate aminotransferase (AATase) to examine the presence of putative excitatory amino acid transmitters in neurons of the deep cerebellar nuclei. Carbodiimide-fixed glutamate-like, GLNase-like and AATase-like immunoreactivities were observed in neurons of the lateral, posterior interpositus, anterior interpositus and medial deep cerebellar nuclei. More neurons were stained with AATase antiserum than with the GLNase antiserum or the monoclonal antibody. These results suggest glutamate, GLNase and AATase are present in neurons of the deep cerebellar nuclei and raise the possibility that glutamate may be an excitatory transmitter in these structures.

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Immunohistochemical localization of glutamate, glutaminase and aspartate aminotransferase in neurons of the pontine nuclei of the rat.

The pontine nuclei form the key relay nuclei in the cerebropontocerebellar pathway. Although a great deal of information is available regarding the anatomy of this region, the identity of the neurotransmitter(s) contained in the neurons of the pontine gray are not known. The aim of the present investigation is to utilize immunohistochemical techniques to determine whether glutamate, a putative excitatory transmitter, and the enzymes responsible for its metabolism, are found in pontine neurons. Both glutaminase, an enzyme which converts glutamine to glutamate, and aspartate aminotransferase, an enzyme which is involved in the interconversion between glutamate and aspartate, have been proposed to be markers of neurons which use excitatory amino acids as neurotransmitters. The present study utilizes a monoclonal antibody against carbodiimide-fixed glutamate and polyclonal antisera against glutaminase and aspartate aminotransferase in conjunction with the indirect peroxidase technique or the peroxidase-labeled biotin-avidin procedure to localize glutamatergic neurons in the pontine nuclei of the rat. Numerous neurons in all subdivisions of the pontine nuclei were found to contain carbodiimide-fixed glutamate-like immunoreactivity, glutaminase-like immunoreactivity or aspartate aminotransferase-like immunoreactivity. Horseradish peroxidase was injected into the cerebellum of four rats for use with a combined retrograde transport-immunohistochemical procedure. Double-labeled neurons were observed in all subdivisions of the pontine nuclei, indicating that pontine neurons which contain glutamate-like immunoreactivity project to the cerebellum. Based on the hypothesis that increased levels of glutamate, glutaminase and aspartate aminotransferase reflect a transmitter role for glutamate, the present data raise the possibility that glutamate may be a major neurotransmitter of pontocerebellar fibers.

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Monoclonal antibody specific for carbodiimide-fixed glutamate: immunocytochemical localization in the rat CNS.

Glutamate is widely distributed in the central nervous system (CNS) and is present in greater amounts than any other putative neurotransmitter. To study its distribution in the CNS, a monoclonal antibody was raised against gamma-L-glutamyl-L-glutamic acid (gamma-Glu-Glu) conjugated to keyhole limpet hemocyanin (KLH) using glutaraldehydeborohydride. By use of this antibody, indirect immunoperoxidase staining was observed in CNS tissue fixed with carbodiimide to form gamma-Glu-Glu from glutamate and post-fixed with glutaraldehyde or paraformaldehyde. In contrast, immunoreactivity was quite low in tissues fixed only with glutaraldehyde. Absorption controls indicated that the staining of carbodiimide-fixed tissue could be inhibited by micromolar concentrations of gamma-Glu-Glu but not by other small molecules. Using ELISA, the antibody reacted strongly with the gamma-Glu-Glu/KLH conjugate used to immunize the mouse, but not with other small molecules conjugated to KLH. The reactivity of the antibody with the gamma-Glu-Glu/KLH conjugate on ELISA was inhibited by free gamma-Glu-Glu in micromolar concentrations, but not by similar dipeptides or amino acids. Dense immunocytochemical staining was observed in cortical pyramidal cells, cerebellar granule cells, and the cochlear nuclei. Staining with this monoclonal antibody correlated well with other methods of localizing glutamate in the CNS.

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Autoradiographic localization of tryptamine binding sites in the rat and dog central nervous system.

Tryptamine, an endogenous trace amine, is currently postulated to be a neuromodulator or neurotransmitter in the mammalian CNS. High-affinity binding sites have been described for tryptamine in rat brain homogenate preparations. The present study further characterizes tryptamine binding throughout the CNS and delineates its distribution using in vitro receptor binding in conjunction with autoradiographic techniques. Saturation studies on 20-micron-thick brain sections suggest a single class of binding sites (Hill coefficient = 0.97 +/- 0.04) with a high affinity (KD = 4.79 +/- 1.55 nM). In competition studies, kynuramine and tetrahydrobetacarboline significantly inhibited H3-tryptamine binding while serotonin, dopamine, and phenylethylamine failed to significantly inhibit it. The most potent inhibitor of H3-tryptamine binding was tryptamine (KI = 4.19 +/- 2.13 nM). In rat brain sections processed for in vitro autoradiography, highest binding occurred in the following limbic structures: the accumbens nucleus, the amygdalohippocampal area, the lateral septal nucleus, the entorhinal cortex, and the anterior olfactory nucleus. At diencephalic levels, the highest binding was observed in the reuniens thalamic nucleus, the paraventricular thalamic nucleus, the medial habenular nucleus, the central medial thalamic nucleus, and the arcuate hypothalamic nucleus. In the midbrain of the rat, binding was most notable in the interpeduncular nucleus, the superficial layer of the superior colliculus, the periaqueductal gray, and the paranigral nucleus. In the lower brain stem of the dog, binding was evident in the external cuneate nucleus, the spinal trigeminal nucleus, and in the region of the solitary nucleus. Binding was also present in both the ventral and dorsal horns of the canine spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS)

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Inhibition of intrathecally administered picrotoxin- and bicuculline-induced convulsions in mice by pipecolic acid or GABA.

Pipecolic acid (PA) is an alicyclic amino acid and putative neurotransmitter which may modulate GABAergic transmission in the central nervous system. The present study was designed to investigate the anticonvulsant effect of intrathecally (i.t.) injected PA on picrotoxin- and bicuculline-induced convulsions which are thought to be produced by interactions with GABAergic systems. Intrathecal injections of picrotoxin and bicuculline in mice produced convulsions which were characterized by a rapid onset and short duration. Coadministration of GABA with either bicuculline or picrotoxin, but not strychnine, attenuated the severity of the convulsions. Coadministration of PA also protected against bicuculline- and picrotoxin-induced convulsions. Intrathecal injections of PA produced a dose-related increase in the latency to the onset of these convulsions as well as a decrease in their duration, however PA failed to inhibit the duration of strychnine-induced seizures. The D isomer of PA was found to be more effective than the L isomer as an anticonvulsant in this study. When administered in a high dose (500 micrograms i.t.), the D isomer produced flaccid paralysis while injection of high doses (100-500 micrograms i.t.) of the L isomer actually elicited convulsions. These results further support an interaction between PA and GABAergic activity.

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The midbrain periaqueductal gray in the rat. I. Nuclear volume, cell number, density, orientation, and regional subdivisions.

The midbrain periaqueductal gray is a functionally heterogeneous region which plays an important role in pain modulation. Despite the heterogeneity considerable controversy exists regarding the presence or absence of morphological subdivisions within the region. The present study was designed to evaluate the possibility of morphological subdivisions within the rat periaqueductal gray by using a statistical cluster analysis system. In addition both qualitative and quantitative data concerning neuronal size, shape, and density were obtained. On the basis of measurements of over 12,000 neurons in two planes of section, the mean neuronal length of cell bodies in this region was 14.82 microns and the mean neuronal area was 95.59 microns squared . The mean neuronal density was found to be 16,284 cells per mm3. Neuronal density decreased from rostral to caudal in the periaqueductal gray. The data obtained from cluster maps suggest the presence of four subdivisions within this midbrain region. The medial subdivision contains the smallest neurons and exhibits the lowest cell density. The dorsolateral and ventrolateral divisions contain the largest neurons while the dorsal division displays the highest packing density. These results are discussed in light of recent receptor binding and immunohistochemical studies of this region.

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The midbrain periaqueductal gray in the rat. II. A Golgi analysis.

This study consists of a detailed analysis of neurons in the midbrain periaqueductal gray of the rat utilizing four variants of the Golgi technique. Neurons were classified into three major categories based on soma shape, number of primary dendrites, number of dendritic bifurcations, interspinous distance, axonal origin, and axon trajectory. Neurons in each category were further subdivided into large and small varieties based predominantly on soma size and dendritic patterns. Both quantitative and qualitative data concerning each neuronal type is provided as well as data relating to its relative distribution among the four periaqueductal gray subdivisions. The small bipolar neuron, characterized by its small size and spindle-shaped soma, was the most prominent cell type observed, composing 37% of the impregnated neurons in our material. This cell type was most numerous in the medial subdivision and least prominent in the dorsolateral subdivision. The small triangular neuron composed 23% of the neuronal population and was relatively evenly distributed through the periaqueductal gray. The remaining four cell types include the large and small multipolar neurons, the large fusiform neurons, and the large triangular neurons. Axons originated from either the perikaryon or a proximal dendrite, with a dendritic origin being most common for large and small triangular neurons and large fusiform neurons. The trajectory of axons in single thick coronal sections originating from periaqueductal gray neurons is typically away from the mesencephalic aqueduct. The exact trajectory is dependent on the location of the neuron. Axons arising from cells in the dorsal subdivision usually project in a dorsal or dorsolateral direction while axons of ventrolateral neurons may project dorsally, laterally, or ventrally. In sum, these data indicate a complex level of internal organization of the periaqueductal gray. The results are discussed in terms of previous immunohistochemical studies of neurons in this region.

Age Factors↗

Immunocytochemical localization of serotonin in the rat periaqueductal gray: a quantitative light and electron microscopic study.

The distribution of serotonin-like immunoreactivity in five regions of the rodent midbrain periaqueductal gray (PAG) was studied by using light and electron microscopic immunohistochemistry in combination with quantitative analysis. Light microscopic analysis revealed the presence of serotonin-like immunoreactive cell bodies located in the ventrolateral and ventromedial regions of the caudal PAG and serotonin-like immunoreactive processes throughout the PAG. Ultrastructural analysis showed dendritic profiles that stained positively for serotonin primarily in ventral regions, although an occasional profile was seen dorsally. Numerous synaptic contacts between unstained axon terminals and ventral dendritic profiles were seen. Axonal profiles that contained reaction product were identified throughout the PAG, but were rarely observed to make any type of specialized contact. Ultrastructural quantification of serotonin-like immunoreactive processes indicated that the highest volume fraction of serotonin immunoreactivity occurred caudoventrally where stained processes constituted 2.6% of the neuropil volume. Rostroventrally stained processes constituted only 0.14% of the neuropil volume at the level of the posterior commissure. By contrast the amount of serotonin-like immunoreactivity found dorsally remained relatively constant at all rostrocaudal levels. Analysis of serotonin staining among PAG regions demonstrated the lowest overall volume fraction in the dorsal region and the highest overall volume fraction in the ventromedial region. No significant differences were observed between medial and lateral regions. A comparison of the results of light microscopic quantitative analysis of serotoninergic processes with electron microscopic quantitative analysis indicated that both techniques produce comparable results.

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The effects of different pretreatment conditions and fixation regimes on serotonin immunoreactivity: a quantitative light microscopic study.

An investigation was designed to evaluate the effects of three different fixation regimes on the retention of serotonin-like immunoreactivity in rat midbrain tissue sections. The effects of pretreatment with pargyline-HCl and l-tryptophan on the volume fraction of serotonin-like immunoreactive processes were also examined. Rat brain tissue was fixed with 4% paraformaldehyde (Pf), 4% paraformaldehyde-0.2% picric acid-0.05% glutaraldehyde (Pf-Pa-G), or 4% paraformaldehyde-0.2% glutaraldehyde (Pf-G). Tissue was subsequently processed for immunohistochemistry using a modified peroxidase-antiperoxidase technique and quantified at the light microscopic level by point counting. Fixation with Pf resulted in higher volume fraction determinations of axonal serotonin immunoreactivity than did fixation with Pf-Pa-G or Pf-G. These results provide quantitative data which indicate that even low levels of glutaraldehyde in the fixative significantly decrease serotonin immunoreactivity. Pretreatment with pargyline and tryptophan increased the amount of serotonin immunoreactivity in tissue fixed with Pf-G but not in tissue fixed with Pf. Pretreatment with pargyline and tryptophan is thus recommended when using glutaraldehyde in the fixation process to assure adequate serotonin immunoreactivity. Pretreatment in conjunction with glutaraldehyde fixation, however, appears to cause differential increases in serotonin-like immunoreactivity within brain nuclei that may compromise the interpretation of results.

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Muscarinic cholinergic receptors in the rat deep cerebellar nuclei: a quantitative autoradiographic study.

The distribution of muscarinic cholinergic receptors within the rat deep cerebellar nuclei was analyzed using in vitro receptor binding of [3H]quinuclidinylbenzilate (QNB) in conjunction with autoradiography. The highest density of QNB binding sites occurred in the lateral cerebellar (dentate) nucleus. Interpositus nuclei displayed an intermediate density of muscarinic cholinergic binding sites with the posterior interpositus nucleus demonstrating higher binding than the anterior nucleus. The fastigial (medial) cerebellar nucleus exhibited the lowest levels of QNB binding among the four cerebellar nuclei. These results indicate that muscarinic cholinergic receptors are present in the deep cerebellar nuclei and that differences in receptor density occur among the four nuclear groups.

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The periaqueductal gray projections to the rat spinal trigeminal, raphe magnus, gigantocellular pars alpha and paragigantocellular nuclei arise from separate neurons.

Possible collateral branches of periaqueductal gray axons which distribute to the nucleus raphe magnus, nucleus reticularis paragigantocellularis, nucleus reticularis gigantocellularis pars alpha and the spinal trigeminal nucleus were analyzed with the double fluorescent retrograde tracer technique. With the exception of a small number of double-labeled neurons observed in the periaqueductal gray following injections of fluorescent dyes into the nuclei reticularis paragigantocellularis and gigantocellularis pars alpha, no double-labeled cells were found in this midbrain region following injections of tracers into various combinations of the above 4 nuclear groups. The results of this investigation indicate that these 4 brainstem nuclei are innervated predominantly by separate neuronal populations within the periaqueductal gray.

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The periaqueductal gray-raphe magnus projection contains somatostatin, neurotensin and serotonin but not cholecystokinin.

The retrograde transport-HRP-immunocytochemical technique was employed to ascertain if the periaqueductal gray-raphe magnus projection arises from neurons containing somatostatin, neurotensin, serotonin or cholecystokinin. Following HRP injections into the raphe magnus (NRM) double-labeled cells containing HRP reaction product and somatostatin-, neurotensin- or serotonin-like immunoreactivity were identified in the midbrain periaqueductal gray (PAG). No cholecystokinin-like immunoreactive double-labeled neurons were found in the PAG. These results indicate that the PAG-NRM pathway contains somatostatin, neurotensin and serotonin but not cholecystokinin.

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The location of brainstem neurons which project bilaterally to the spinal trigeminal nuclei as demonstrated by the double fluorescent retrograde tracer technique.

Cells with possible dual projections to both spinal trigeminal nuclei were identified in the rat brainstem following separate injections of different retrogradely transported markers into the right and left spinal trigeminal nucleus. The greatest number of double-labeled cells was located in the nucleus reticularis gigantocellularis. Several double-marked cells were also observed in the nucleus raphe magnus, the nucleus paragigantocellularis and the periaqueductal gray. These results suggest that some cells in the above brainstem nuclei may have a bilateral modulating effect on the spinal trigeminal nuclei.

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