Organization of choline acetyltransferase-containing structures in the cranial nerve motor nuclei and lamina IX of the cervical spinal cord of the rat

The cholinergic structures in the cranial nerve motor nuclei and lamina IX of the cervical spinal cord of the rat were examined immunohistochemically with a monoclonal antibody to choline acetyltransferase (ChAT). The brainstem motor nuclei were classified into 3 groups according to the way of distribution of ChAT-positive structures in the neuropil of the nucleus. 1. The oculomotor, trochlear and abducent nuclei contained moderately ChAT-positive perikarya and dendrites. A small number of ChAT-positive bouton-like structures were found in the neuropil, but they were not in contact with ChAT-positive perikarya and dendrites. 2. Motoneurons in the facial, hypoglossal and trigeminal motor nuclei were moderately to strongly ChAT-positive. There were in the neuropil numerous ChAT-positive bouton-like structures and many of them were in contact with ChAT-positive perikarya and dendrites. The same pattern of organization of ChAT-positive structures was observed in lamina IX of the cervical spinal cord. In the nucleus ambiguus, the perikarya and proximal dendrites of about half population of motoneurons contacted with ChAT-positive bouton-like structures while the rest of motoneurons did not. ChAT-positive bouton-like structures in contact with motoneurons are interpreted as the cholinergic synaptic terminals, and this suggests that motoneurons in this group are cholinoceptive as well as cholinergic. 3. Neuronal perikarya and dendrites in the dorsal nucleus of the vagus showed weak to moderate positivity of ChAT. The neuropil of this nucleus was free from any distinctly ChAT-positive structures.     

Distribution of high affinity choline transporter immunoreactivity in the primate central nervous system

A mouse monoclonal antibody (clone 62-2E8) raised against a human recombinant high-affinity choline transporter (CHT)-glutathione-S-transferase fusion protein was used to determine the distribution of immunoreactive profiles containing this protein in the monkey central nervous system (CNS). Within the monkey telencephalon, CHT-immunoreactive perikarya were found in the striatum, nucleus accumbens, medial septum, vertical and horizontal limb nuclei of the diagonal band, nucleus basalis complex, and the bed nucleus of the stria terminalis. Dense fiber staining was observed within the islands of Calleja, olfactory tubercle, hippocampal complex, amygdala; moderate to light fiber staining was seen in iso- and limbic cortices. CHT-containing fibers were also present in sensory and limbic thalamic nuclei, preoptic and hypothalamic areas, and the floccular lobe of the cerebellum. In the brainstem, CHT-immunoreactive profiles were observed in the pedunculopontine and dorsolateral tegmental nuclei, the Edinger-Westphal, oculomotor, trochlear, trigeminal, abducens, facial, ambiguus, dorsal vagal motor, and hypoglossal nuclei. In the spinal cord, CHT-immunoreactive ventral horn motoneurons were seen in close apposition to intensely immunoreactive C-terminals at the level of the cervical spinal cord. CHT immunostaining revealed a similar distribution of labeled profiles in the aged human brain and spinal cord. Dual fluorescent confocal microscopy revealed that the majority of CHT immunoreactive neurons contained the specific cholinergic marker, choline acetyltransferase, at all levels of the monkey CNS. The present observations indicate that the present CHT antibody labels cholinergic structures within the primate CNS and provides an additional marker for the investigation of cholinergic neuronal function in aging and disease.     

Oculomotor system of the weakly electric fish Gnathonemus petersii

The peripheral and central aspects of the extraocular system were studied in the weakly electric fish Gnathonemus petersii. All six extraocular muscles show a similar composition of large and small fibers grouped characteristically in the proximal and distal regions respectively. The exit of the three extraocular nerves from the brain is similar to that in other vertebrates. However, the intracephalic and intracranial course of the trochlear nerve is unusual, partly because of the extraordinary hypertrophy of the cerebellum. The three nerves course rostrally on the ventral brain surface; the trochlear nerve penetrates the orbital cavity separately from the two other nerves. The fiber-diameter spectrum of each extraocular nerve is bimodal; unmyelinated fibers were not observed in any of the nerves. The location of the extraocular motor nuclei was established by retrograde axonal transport of HRP or cobaltic-lysine complex. The oculomotor nucleus is situated ventral to the posterior pole of the magnocellular mesencephalic nucleus and the trochlear nucleus is found caudal and dorsal to this. The abducens nucleus is situated at the level of the octavolateral efferent nucleus and consists of a single group of cells on each side of the ventral tegmentum. The oculomotor nucleus of G. petersii shows a somatotopic organization. The superior rectus muscle receives a contralateral innervation whereas the inferior rectus and oblique muscles and the internal rectus muscles receive an ipsilateral innervation. The superior oblique muscle is innervated by contralateral trochlear motoneurons and the external rectus by ipsilateral abducens motoneurons. The majority of extraocular motoneurons have piriform perikarya and long beaded dendrites that extend laterally in the oculomotor and abducens nuclei and rostrally in the trochlear nucleus. The terminal dendritic portions of trochlear motoneurons widely overlap with oculomotor dendrites and perikarya. In all three nuclei the axon originates opposite to the main dendrite. Collaterals of the hairpin-bend abducens axons could be identified in a few cases. The oculomotor system of G. petersii appears basically similar to that of other teleosts; the differences observed concern mainly the structure of the abducens nucleus, the intracranial and intracephalic course of the trochlear nerve, and the relatively small number of axons in each nerve.     

Brainstem motoneuron pools that are selectively resistant in amyotrophic lateral sclerosis are preferentially enriched in parvalbumin: Evidence from monkey brainstem for a calcium-mediated mechanism in sporadic ALS

Some brainstem motoneuron groups appear more resistant to the process of neurodegeneration in ALS (for example, oculomotor, trochlear, and abducens nuclei) than others (for example, trigeminal, facial, ambiguus, and hypoglossal nuclei). The possibility that the differential presence of the calcium-chelating protein parvalbumin might underlie this difference in vulnerability was examined immunohistochemically as a way to determine whether a calcium-mediated mechanism might be involved in ALS. In normal monkey brainstem, we found that the abundance of parvalbumin-containing neurons in the oculomotor, trochlear, and abducens nuclei was approximately 90% of the abudance of choline acetyltransferase (CHAT)-containing motoneurons. In contrast, the abundance of parvalbumin-containing neurons in the other brainstem motor nuclei innervating skeletal muscle (trigeminal, facial, ambiguus, and hypoglossal) was only about 30–60% of the abundance of CHAT-containing motoneurons. Since some of these motoneuron pools contain nonmotoneuron internuclear neurons that might be parvalbumin-containing, we also carried out double-label studies to specifically determine the percentage of cholinergic motoneurons that contained paravalbumin in each of these motoneuron pools. We found that 85–100% of the oculomotor, trochlear, and abducens motoneurons were parvalbumin-containing. In contrast, only 20–30% of the trigeminal, facial, ambiguus, and hypoglossal motoneurons were parvalbumin-containing. These results raise the possibility that motoneuron death in sporadic ALS is related to some defect that promotes cytosolic calcium accumulation in motoneurons. This excess calcium entry may promote cell death via an excitotoxic pathway. Motoneurons rich in parvalbumin may resist the deleterious effects of this putative calcium gating defect because they are better able to sequester the excess calcium.     

In situ hybridization localization of choline acetyltransferase mRNA in adult rat brain and spinal cord

The cellular distribution of choline acetyltransferase (ChAT) mRNA within the adult rat central nervous system was evaluated using in situ hybridization. In forebrain, hybridization of a ³⁵S-labeled rat ChAT cRNA densely labeled neurons in the well-characterized basal forebrain cholinergic system including the medial septal nucleus, diagonal bands of Broca, nucleus basalis of Meynert and substantia innominata, as well as in the striatum, ventral pallidum, and olfactory tubercle. A small number of lightly labeled neurons were distributed throughout neocortex, primarily in superficial layers. No cellular labeling was detected in hippocampus. In the diencephalon, dense hybridization labeled neurons in the ventral aspect of the medial habenular nucleus whereas cells in the lateral hypothalamic area and supramammillary region were more lightly labeled. Hybridization was most dense in neurons of the motor and autonomic cranial nerve nuclei including the oculomotor, Edinger-Westphal, and trochlear nuclei of the midbrain, the abducens, superior salivatory, trigeminal, facial and accessory facial nuclei of the pons, and the hypoglossal, vagus, and solitary nuclei and nucleus ambiguus of the medulla. In addition, numerous cells in the pedunculopontine and laterodorsal tegmental nuclei, the ventral nucleus of the lateral lemniscus, the medial and lateral divisions of the parabrachial nucleus, and the medial and lateral superior olive were labeled. Occasional labeled neurons were distributed in the giantocellular, intermediate, and parvocellular reticular nuclei, and the raphe magnus nucleus. In the medulla, light to moderately densely labeled cells were scattered in the nucleus of Probst's bundle, the medial vestibular nucleus, the lateral reticular nucleus, and the raphe obscurus nucleus. In spinal cord, the cRNA densely labeled motor neurons of the ventral horn, and cells in the intermediolateral column, surrounding the central canal, and in the spinal accessory nucleus. These results are in good agreement with reports of the immunohistochemical localization of ChAT and provide further evidence that cholinergic neurons are present within neocortex but not hippocampus.     

Expression of a serine protease (motopsin PRSS12) mRNA in the mouse brain: in situ hybridization histochemical study

Serine proteases are considered to play several important roles in the brain. In an attempt to find novel brain-specific serine proteases (BSSPs), motopsin (PRSS-12) was cloned from a mouse brain cDNA library by polymerase chain reaction (PCR). Northern blot analysis demonstrated that the postnatal 10-day mouse brain contained the most amount of motopsin mRNA. At this developmental stage, in situ hybridization histochemistry showed that motopsin mRNA was specifically expressed in the following regions: cerebral cortical layers II/III, V and VIb, endopiriform cortex and the limbic system, particularly in the CA1 region of the hippocampal formation. In addition, in the brainstem, the oculomotor nucleus, trochlear nucleus, mecencephalic and motor nuclei of trigeminal nerve (N), abducens nucleus, facial nucleus, nucleus of the raphe pontis, dorsoral motor nucleus of vagal N, hypoglossal nucleus and ambiguus nucleus showed motopsin mRNA expression. Expression was also found in the anterior horn of the spinal cord. The above findings strongly suggest that neurons in almost all motor nuclei, particularly in the brainstem and spinal cord, express motopsin mRNA, and that motopsin seems to have a close relation to the functional role of efferent neurons.     

Intraneuronal IgG in the central nervous system

The rat central nervous system was examined immunocytochemically for the presence of endogenous IgG. Examination of representative sections of the neuraxis revealed specific staining for IgG in the pia mater and pial vasculature, the ependyma, and diffusely in the hypothalamus and area postrema where the blood-brain barrier is permeable to large molecules. In addition, intraneuronal staining for IgG was noted in specific nuclei including the ventral horn nuclei and intermediolateral nuclei of the spinal cord, the dorsal motor nucleus of the vagus, the nucleus ambiguous, the motor nucleus of the trigeminal, the hypoglossal, facial, and oculomotor nuclei, nuclei projecting to the pituitary and area postrema, and Purkinje cells. The uptake of immunoglobins by these cell groups may have important implications for the pathogenesis of motor and autonomic neuropathies and neuropathies.     

Extra- and intracellular HRP analysis of the organization of extraocular motoneurons and internuclear neurons in the guinea pig and rabbit

The distribution of extraocular motoneurons and abducens and oculomotor internuclear neurons was determined in guinea pigs by injecting horseradish peroxidase (HRP) into individual extraocular muscles, the abducens nucleus, the oculomotor nucleus, and the cerebellum. Motoneurons in the oculomotor nucleus innervated the ipsilateral inferior rectus, inferior oblique, medial rectus, and the contralateral superior rectus and levator palpebrae muscles. Most motoneurons of the trochlear nucleus projected to the contralateral superior oblique muscle although a small number innervated the ipsilateral superior oblique. The abducens and accessory abducens nuclei innervated the ipsilateral rectus and retractor bulbi muscles, respectively. The somata of abducens internuclear neurons formed a cap around the lateral and ventral aspects of the abducens nucleus. The axons of these internuclear neurons terminated in the medial rectus subdivision of the contralateral oculomotor nucleus. At least two classes of guinea pig oculomotor internuclear interneurons exist. One group, located primarily ventral to the oculomotor nucleus, innervated the abducens nucleus and surrounding regions. The second group, lying mainly in the dorsal midline area of the oculomotor nucleus, projected to the cerebellum. Intracellular staining with HRP demonstrated similar soma-dendritic organization for oculomotor and trochlear motoneurons of both guinea pigs and rabbits. Dendrites of oculomotor motoneurons radiated symmetrically from the soma to cover approximately one-third of the entire nucleus, and each motoneuron sent at least one dendrite into the central gray overlying the oculomotor nucleus. In both species, a small percentage of oculomotor motoneurons possessed axon collaterals that terminated both within and outside of the nucleus. The dendrites of trochlear motoneurons extended into the medial longitudinal fasciculus and the reticular formation lateral to the nucleus. Our data on the topography of motoneurons and internuclear neurons in the guinea pig and soma-dendritic organization of motoneurons in the guinea pig and rabbit show that these species share common organizational and morphological features. In addition, comparison of these data with those from other mammals reveals that dendritic complexity (number of dendrites per motoneuron) of extraocular motoneurons exhibits a systematic increase with animal size.     

Distribution of acidic fibroblast growth factor mRNA-expressing neurons in the adult mouse central nervous system

The distribution of acidic fibroblast growth factor (aFGF) mRNA-expressing neurons was studied throughout the adult mouse central nervous system (CNS) with in situ hybridization histochemistry using a radiolabelled synthetic oligodeoxynucleotide probe complementary to the mRNA of human aFGF. We report here a widespread distribution of aFGF mRNA in several defined functional systems of the adult mouse brain, whereby the highest levels of aFGF mRNA were found in large somatomotor neurons in the nuclei of the oculomotor, trochlear, abducens, and hypoglossal nerves; in the motoneurons of the ventral spinal cord and the special visceromotor neurons in the motor nucleus of the trigeminal nerve; and in the facial and ambigaus nuclei. Labelled perikarya were also detected in all central structures of the auditory pathway including the level of the inferior colliculus, i.e., the lateral and medial superior nuclei; the trapezoid, cochlear, and lateral lemniscal nuclei; and parts of the anterior colliculus. Furthermore, many aFGF-positive cell bodies were found in the vestibular system and other structures projecting to the cerebellum, in the deep cerebellar nuclei, in somatosensory structures of the medulla (i.e., in the gracile, cuneate, and external cuneate nuclei), as well as in the spinal nucleus of the trigeminal nerve. The findings that aFGF mRNA is expressed in all components of several well-defined systems (i.e., in sensory structures) as Well as in central neurons that process sensory information and, finally, in some efferent projections point towards a concept of aFGF expression primarily within certain neuronal circuitries. © 1995 Wiley-Liss, Inc.     

Dendrodendritic and dendrosomatic contacts between oculomotor and trochlear motoneurons of the frog, Rana esculenta

Gaze fixation requires very fast movements of the eye during body displacement. The morphological and physiological background of the very fine and continuous tuning of gaze fixation is not yet fully understood. In a previous study we have shown that the dendrites of oculomotor neurons form bundles which invade the trochlear nucleus, and vice versa, trochlear dendritic bundles invade the oculomotor nucleus. Earlier physiological observations demonstrating electrotonic coupling between dendrites of spinal motoneurons in the frog suggest a similar mechanism between the oculomotor and trochlear motoneurons. We studied a possible morphological basis of gaze fixation. The experiments were carried out on common water frogs, Rana esculenta. The trochlear and oculomotor nerves were cut, and their proximal stumps were labeled simultaneously with different retrograde fluorescent tracers. Using confocal laser scanning microscope we detected a large number of close contacts in both nuclei, the majority of them were dendrodendritic apposition. The distance between the adjacent profiles suggested close membrane appositions without intercalating glial or neuronal elements. At the ultrastructural level, the dendrodendritic and dendrosomatic contacts did not show any morphological specialization; the long membrane appositions may provide ephaptic interactions between the neighboring profiles. This electrotonic coupling between the oculomotor and trochlear nerve motoneurons may promote the co-activation of the muscles responsible for vertical eye movements.     

Dendrodendritic and dendrosomatic contacts between oculomotor and trochlear motoneurons of the frog, Rana esculenta

Gaze fixation requires very fast movements of the eye during body displacement. The morphological and physiological background of the very fine and continuous tuning of gaze fixation is not yet fully understood. In a previous study we have shown that the dendrites of oculomotor neurons form bundles which invade the trochlear nucleus, and vice versa, trochlear dendritic bundles invade the oculomotor nucleus. Earlier physiological observations demonstrating electrotonic coupling between dendrites of spinal motoneurons in the frog suggest a similar mechanism between the oculomotor and trochlear motoneurons. We studied a possible morphological basis of gaze fixation.The experiments were carried out on common water frogs, Rana esculenta. The trochlear and oculomotor nerves were cut, and their proximal stumps were labeled simultaneously with different retrograde fluorescent tracers. Using confocal laser scanning microscope we detected a large number of close contacts in both nuclei, the majority of them were dendrodendritic apposition. The distance between the adjacent profiles suggested close membrane appositions without intercalating glial or neuronal elements. At the ultrastructural level, the dendrodendritic and dendrosomatic contacts did not show any morphological specialization; the long membrane appositions may provide ephaptic interactions between the neighboring profiles. This electrotonic coupling between the oculomotor and trochlear nerve motoneurons may promote the co-activation of the muscles responsible for vertical eye movements.     

Afferent projections to the oral motor nuclei in the rat

Projections to the trigeminal, facial, ambiguus, and hypoglossal motor nuclei were determined by using horseradish peroxidase histochemistry. Most of the afferent projections to these motor nuclei were from the brainstem reticular formation, frequently in areas adjacent to other synergetic motor nuclei. The reticular formation lateral to the hypoglossal nucleus and reticular structures surrounding the trigeminal motor nucleus projected to each of these other brainstem motor nuclei involved in oral-facial function. Afferent projections to these motor nuclei also were organized along the rostrocaudal axis. Within the reticular formation most of the afferent projections to the trigeminal motor nucleus originated rostral to the majority of neurons projecting to the hypoglossal and ambiguus nuclei, which in turn were rostral to the primary source of reticular afferents to the facial nucleus. In comparison, projections from the sensory trigeminal nuclei and nucleus of the solitary tract were sparse. The interneuron pools that project to the orofacial motoneurons provide one further link in understanding the brainstem substrates for integrating oral and ingestive behaviors.     

A systematic study of brainstem motor nuclei in a mouse model of ALS,the effects of lithium

Transgenic mice expressing the human superoxide dismutase 1 (SOD-1) mutant at position 93 (G93A) develop a phenotype resembling amyotrophic lateral sclerosis (ALS). In fact, G93A mice develop progressive motor deficits which finally lead to motor palsy and death. This is due to the progressive degeneration of motor neurons in the ventral horn of the spinal cord. Although a similar loss is reported for specific cranial motor nuclei, only a few studies so far investigated degeneration in a few brainstem nuclei. We recently reported that chronic lithium administration delays onset and duration of the disease, while reducing degeneration of spinal motor neuron. In the present study, we extended this investigation to all somatic motor nuclei of the brain stem in the G93A mice and we evaluated whether analogous protective effects induced by lithium in the spinal cord were present at the brain stem level. We found that all motor but the oculomotor nuclei were markedly degenerated in G93A mice, and chronic treatment with lithium significantly attenuated neurodegeneration in the trigeminal, facial, ambiguus, and hypoglossal nuclei. Moreover, in the hypoglossal nucleus, we found that recurrent collaterals were markedly lost in G93A mice while they were rescued by chronic lithium administration.     

Motoneurons of twitch and nontwitch extraocular muscle fibers in the abducens, trochlear, and oculomotor nuclei of monkeys

Eye muscle fibers can be divided into two categories: nontwitch, multiply innervated muscle fibers (MIFs), and twitch, singly innervated muscle fibers (SIFs). We investigated the location of motoneurons supplying SIFs and MIFs in the six extraocular muscles of monkeys. Injections of retrograde tracers into eye muscles were placed either centrally, within the central SIF endplate zone; in an intermediate zone, outside the SIF endplate zone, targeting MIF endplates along the length of muscle fiber; or distally, into the myotendinous junction containing palisade endings. Central injections labeled large motoneurons within the abducens, trochlear or oculomotor nucleus, and smaller motoneurons lying mainly around the periphery of the motor nuclei. Intermediate injections labeled some large motoneurons within the motor nuclei but also labeled many peripheral motoneurons. Distal injections labeled small and medium-large peripheral neurons strongly and almost exclusively. The peripheral neurons labeled from the lateral rectus muscle surround the medial half of the abducens nucleus: from superior oblique, they form a cap over the dorsal trochlear nucleus; from inferior oblique and superior rectus, they are scattered bilaterally around the midline, between the oculomotor nucleus; from both medial and inferior rectus, they lie mainly in the C-group, on the dorsomedial border of oculomotor nucleus. In the medial rectus distal injections, a "C-group extension" extended up to the Edinger-Westphal nucleus and labeled dendrites within the supraoculomotor area. We conclude that large motoneurons within the motor nuclei innervate twitch fibers, whereas smaller motoneurons around the periphery innervate nontwitch, MIF fibers. The peripheral subgroups also contain medium-large neurons which may be associated with the palisade endings of global MIFs. The role of MIFs in eye movements is unclear, but the concept of a final common pathway must now be reconsidered.     

Ascending and descending projections from nucleus reticularis magnocellularis and nucleus reticularis gigantocellularis: an autoradiographic and horseradish peroxidase study in the rat

The projections of the rostal medulla were studied using retrograde and orthograde transport techniques in the rat. The present horseradish peroxidase (HRP) studies indicate that the ventral portion of nucleus reticularis (NGC) and nucleus reticularis magnocellularis (NMC) project to both rostral and caudal levels of the spinal cord, while dorsal NGC projects only to the rostral cord. A differential density distribution of labeled cells was observed, with the greatest density of NGC-spinal neurons located rostral to the level of the inferior olive; and the greatest density of NMC-spinal neurons located caudally.This differential density distribution, when coupled with microiontophoretic application of [³H]amino acids allowed relatively independent labeling of the adjacent NGC- and NMC-spinal systems. On the basis of the HRP and autoradiographic studies 3 separate regions were delineated: dorsal NGC, ventral NGC and NMC. Descending projections from NGC were observed to the lateral vestibular nucleus, facial nucleus, hypoglossal nucleus and cuneatus. At cervical levels NGC fibers projected through the ventral and ventrolateral columns. Terminal fields were observed in laminae VII, VIII and to a lesser extent in IX. Labeled NGC fibers became difficult to follow by thoracic levels, which is consistent with the present HRP results. A continuum of descending NGC projections was observed with dorsally located NGC neurons projecting bilaterally through the ventral columns, and ventrally located NGC cells projecting through the ipsilateral ventrolateral columns. Ascending projections from NGC to the motor nucleus of V, trochlear nucleus, oculomotor nucleus, Edinger-Westphal nucleus, the ventral aspect of the periaqueductal gray, the deep and intermediate layers of the superior colliculus, nucleus parafasicularis and centromedianus, the Fields of Forel and the dorsal and lateral hypothalamic nuclei were observed. Descending projections from NMC to the dorsal nucleus of the vagus, hypoglossal nucleus, nucleus commissuralis and intercalatus were observed. At cervical levels, fibers project through the ipsilateral lateral columns, particularly its dorsal aspect. Terminal fields are located ipsilaterally in lamonae IV, V and VI, and bilaterally in VII, VIII and X. NMC projections continue through cadual levels of the spinal cord including a projection to the ipsilateral intermediolateral columns. Ascending NMC projections are limited to the ventral pontine reticular formation.The differing projections and cytoarchitecture of the rostral medulla of the rat observed in the present study are compared to that of the cat and opossum, with implications for the subdivision of this region discussed. The possible involvement of NMC and NGC projections in the modulation of pain is reviewed.     

A comparative study of the immunohistochemical localization of a presumptive proctolin-like peptide, thyrotropin-releasing hormone and 5-hydroxytryptamine in the rat central nervous system

A proctolin (PROC)-like peptide was studied immunohistochemically in the hypothalamus, lower brainstem and spinal cord of the rat using an antiserum against PROC conjugated to thyroglobulin. Neuronal cell bodies containing PROC-like immunoreactivity (PROC-LI) were observed in the dorsomedial, paraventricular and supraoptic nuclei of the hypothalamus and in the nucleus raphe magnus, nucleus raphe pallidus, nucleus raphe obscurus and nucleus interfascicularis nervi hypoglossi in the medulla oblongata. Fibers containing PROC-LI were seen in the median eminence and in other hypothalamic nuclei, and in the lower brainstem in cranial motor nuclei including the dorsal motor nucleus of the vagus nerve, the motor trigeminal nucleus, the facial nucleus and nucleus ambiguous, and in lower numbers in the nucleus of the solitary tract and locus coeruleus. Fibers containing PROC-LI were also located in the spinal cord, in the intermediolateral cell column at thoracic levels and in the ventral horns at all levels of the spinal cord. After transection of the spinal cord, all PROC-immunoreactive fibers below the lesion disappeared. Following injection of Fast blue into the thoracic spinal cord, retrogradely labeled cells in the nuclei raphe pallidus, obscurus and magnus and nucleus interfasciculari nervi hypoglossi were seen to contain PROC-LI. PROC-LI had a similar distribution as thyrotropin-releasing hormone (TRH)-LI in the above-mentioned areas and coexistence of TRH-LI and PROC-LI was shown in cell bodies in the hypothalamus and medulla oblongata. PROC-LI could also be shown to coexist with 5-hydroxytryptamine (5-HT)-LI in neuronal cell bodies in the lower brainstem. The results demonstrate the occurrence of a PROC-like peptide in the mammalian nervous system, and these neurons seem to be at least largely identical to previously described TRH systems. A possible involvement of the PROC-like peptide in spinal motor control is discussed in relation to the well-established role of PROC in control of motor behavior in insects and invertebrates.     

Glycogen, its transient occurrence in neurons of the rat CNS during normal postnatal development

Progressive changes in the postnatal incidence, distribution and duration of glycogen in neurons of the pons, medulla and spinal cord were studied by light and electron microscopy using cytochemical and quantitative methods. Albino rats of 11 ages ranging from newborn to adult were used for this investigation. Methacrylate sections, stained with periodic acid-Schiff-dimedone (PAS) were surveyed to identify nerve cell groups containing the polysaccharide, glycogen. The PAS reaction was positive in neuronal cell groups of the hypoglossal nucleus, the mesencephalic nucleus of V, nucleus ambiguus, the abducens nucleus, the facial motor nucleus and anterior horn cells of the spinal cord. The intensity and duration of the PAS reaction appeared greatest in the hypoglossal nucleus. Neurons of the mesencephalic nucleus of V demonstrated a reaction of moderate intensity and duration. The remaining nerve cell groups exhibited a weak, diffuse reaction of brief duration. Postnatal differences in the incidence and patterns of disposition of glycogen were quantified using ultrathin sections of the hypoglossal nucleus, the site richest in glycogen. The presence of glycogen was verified by the periodic acid-thiosemicarbizide-silver proteinate (PA-TSC-SP) ultracytochemical stain. The incidence of glycogen in neuronal perikarya of hypoglossal nuclei was related to age. All neurons contained some glycogen during the first postnatal week. By 24 days postnatal (dpn), the majority of hypoglossal neurons lacked glycogen and all neurons of adult rats were glycogen-free.(ABSTRACT TRUNCATED AT 250 WORDS)     

Localization of choline acetyltransferase in perikarya and dendrites within the nuclei of the solitary tracts

Immunocytochemistry was used to establish the cellular localization of choline acetyltransferase [ChAT] throughout the rostrocaudal portions of the nuclei of the solitary tracts [NTS] in rat brain. By light microscopy, two distinct populations of ChAT-positive cells were identified. The first consisted of relatively few, medium-sized neurons located in the caudal one-half of the medial NTS just dorsal to the dorsal motor nucleus of the vagus. The second population of ChAT-labeled neurons was located more anteriorly and surrounded the medial and dorsal borders of the tractus solitarius. These cells were more abundant and smaller diameter than those located more caudally. Thick, non-varicose processes with the light microscopic characteristics of dendrites also were selectively labeled for ChAT. A few of these processes were located near or were continuous with the labeled perikarya of the NTS. However, the vast majority of the immunoreactive processes could be traced from ChAT-labeled perikarya in the ventrally adjacent dorsal motor nucleus of the vagus. These dorsally directed dendrites aborized extensively throughout the NTS, but they were densest in the rostral two-thirds of the nucleus. Caudally, the labeled dendrites coursed horizontally, forming a commissure-like structure between the two vagal motor nuclei. Electron microscopy confirmed the perikaryal and dendritic localization of ChAT in the NTS. The perikarya were characterized by dense peroxidase immunoreactivity throughout the cytoplasm, infolded nuclear membranes, and somatic synapses. The labeled dendritic profiles also were intensely immunoreactive and received synaptic input from unlabeled terminals. The unlabeled afferents to somata and dendrites contained large populations of small clear vesicles.(ABSTRACT TRUNCATED AT 250 WORDS)