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K Kitahama

Publications and source records attributed to K Kitahama.

At least 55 records · Page 3Linked to original sources

Monoamine oxidase: distribution in the cat brain studied by enzyme- and immunohistochemistry: recent progress.

Localization of MAO-containing neurons, fibers and glial cells has been described by recent progress in MAO histochemistry and immunohistochemistry. It does not necessarily correspond to those containing monoamines. MAO-A is demonstrated in many noradrenergic cells, but it is hardly detectable in DA cells. Increase of 5-HT and DA concentration after inhibition of MAO-A indicates the possible existence of MAO-A in such neuronal structures. MAO-A is also undetectable in neurons containing 5-HT, a good substrate for MAO-A. These neurons contain MAO-B. There still remain contradictions to be solved in future. MAO is present in astroglial cells, in which monoamines released in extracellular space may be degraded. In glial cells, MAO may also play a role to regulate concentration of telemethylhistamine and trace amines. Such cells appear to transform MPTP to MPP+, a neurotoxin for nigral DA neurons.

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Quantitative and qualitative aspects on the distribution of 5-HT and its coexistence with substance P and TRH in cat ventral medullary neurons.

By use of the indirect immunofluorescence technique the distributions of 5-hydroxytryptamine (5-HT)-, substance P- and thyrotropin-releasing hormone (TRH)-immunoreactive (IR) neurons have been studied in the midline raphe nuclei and nucleus reticularis lateralis of the caudal brainstem (levels P18.3-P8.5; according to Berman (1968), in the cat, after treatment with colchicine. In addition, by use of the double-labelling technique, the coexistence between 5-HT-, substance P- and TRH-like immunoreactivity (LI) in these neurons was analysed. The results show that cell bodies in the midline raphe nuclei and nucleus reticularis lateralis contain 5-HT-, substance P- and TRH-LI. 5-HT-IR cells were more abundant than peptidergic neurons in all areas analysed. Quantitative estimations indicated that the total number of 5-HT-IR cells in the regions studied was about 17 x 10(3), while the corresponding numbers for substance P- and TRH-IR cells were 11 x 10(3) and 12 x 10(3), respectively. From double-labelled sections it was concluded that the vast majority of peptidergic cells also contained 5-HT-LI (87-100%). However, a subpopulation of 5-HT-IR neurons lacked peptide-LI (10-55%). The degree of coexistence varied with the brainstem level, in that neurons at more rostral locations showed a lower incidence of coexistence between 5-HT and peptide(s). The presence of all three compounds in one and the same cell body could also be demonstrated. In summary, 5-HT-, substance P- and TRH-IR cell bodies were encountered in medullary nuclei known to contain neurons with projection to the spinal cord. A high degree of coexistence between the compounds was demonstrated in these nuclei. The obtained results fit well with earlier studies on the patterns of distribution and peptide colocalization of 5-HT fibres in the spinal cord. The existence of biochemically distinct neuronal subpopulations within the 5-HT bulbospinal pathway is discussed.

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Adrenergic innervation of the urinary bladder body in the cat with special reference to structure of the detrusor muscle: an immunohistochemical study of noradrenaline and its synthesizing enzymes.

The distribution of adrenergic nerves in the body detrusor muscle of the cat urinary bladder was studied by means of the immunohistochemical identification of noradrenaline (NA) and the NA synthesizing enzymes tyrosine hydroxylase, aromatic L-aminoacid decarboxylase and dopamine beta-hydroxylase. We identified the basic structural organization of the detrusor muscle, which had previously been described as lacking discernible layers. In the lateral wall, both outer longitudinal and inner circular muscle bundles were present, the latter extending in both anterior and posterior directions. The posteriorly running bundles came to lie on the outside of the posterior wall where they enabled recognition of inner longitudinal muscle bundles. Those running anteriorly were dispersed to enter the longitudinal bundles in the anterior wall. NA-immunoreactive nerve fibers in the detrusor muscle of the bladder were found to be similar to those immunoreactive for NA synthesizing enzymes in both distribution and density. In the upper and middle bladder body--including the dome (apex)--immunoreactive nerve fibers were always more abundant in the outer part of the detrusor muscle than in the inner part, regardless of the course of muscle bundles. Even in individual muscle bundles running from the inside to the outer surface, the outer part was more richly innervated by immunoreactive fibers than the inner part. In the bladder dome, a moderate number of immunoreactive nerve fibers preferentially innervated the outer part of the muscle layer. In the lower bladder body, these nerve fibers increased in density in the inner part of the detrusor muscle. There was no sexual difference in density or distribution of nerve fibers. NA- and NA synthesizing enzyme-immunoreactive nerve fibers were markedly decreased in number after 6-hydroxydopamine treatment. No dopamine- or phenylethanolamine-N-methyltransferase-immunoreactive nerve fibers were present in the bladder. The findings of this study indicate that the cat bladder musculature includes longitudinal and circular muscle bundles, both of which are extensively innervated by adrenergic nerves, particularly in the outer part of the bladder.

Adrenergic Fibers↗

Dominant expression of mRNA for prostaglandin D synthase in leptomeninges, choroid plexus, and oligodendrocytes of the adult rat brain.

Glutathione-independent prostaglandin D synthase [prostaglandin-H2 D-isomerase; (5Z,13E)-(15S)-9 alpha,11 alpha-epidioxy-15-hydroxyprosta-5,13-dienoate D-isomerase, EC 5.3.99.2] is an enzyme responsible for biosynthesis of prostaglandin D2 in the central nervous system. In situ hybridization with antisense RNA for the enzyme indicated that mRNA for the enzyme was predominantly expressed in the leptomeninges, choroid plexus, and oligodendrocytes of the adult rat brain. The findings agree with those obtained by immunohistochemical staining with antibodies against the enzyme. It was further revealed that prostaglandin D synthase activity was considerably greater in the isolated leptomeninges (14.2 nmol per min per mg of protein) and choroid plexus (7.0 nmol per min per mg of protein) than the activity in the whole brain (2.0 nmol per min per mg of protein). These results, taken together, indicate that the enzyme is mainly synthesized and located in the leptomeninges, choroid plexus, and oligodendrocytes in the brain.

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Histaminergic system in the cat hypothalamus with reference to type B monoamine oxidase.

It is known that histamine (HA) and type B monoamine oxidase (MAO-B), an enzyme involved in its metabolism, are present in the posterior hypothalamus, but the sites where MAO-B intervenes in HA metabolism remain uncertain. The present study examined and compared the detailed distribution and morphology of neurons immunoreactive to HA (HA-ir) and MAO-B (MAO-B-ir) in the cat hypothalamus. HA-ir neurons were localized almost exclusively in the posterior hypothalamus with the largest group in the tuberomammillary nucleus and adjacent areas. MAO-B-ir staining was detected in the vast majority of HA-ir neurons, suggesting that the degradation of tele-methylhistamine (t-MHA), the direct metabolite of HA, may occur within these cells. Nevertheless, a few HA-ir cells showed no detectable or very weak MAO-B-ir labeling; a small group of neurons containing MAO-B alone was detected in the area dorsolateral to the caudal part of the arcuate nucleus. Numerous HA-ir axons and terminal-like structures were distributed unevenly in virtually all hypothalamic regions. One of their principal trajectories ascended through the ventrolateral part of the hypothalamus and rostrally formed an axon column, which ascended into the preoptic area and contributed fibers to the diagonal band of Broca and bed nucleus of the stria terminalis. Other HA-ir axons passed laterally, dorsal to the zona incerta or ventrally through a narrow zone dorsal to the optic tract. Numerous long HA-ir axons coursed dorsomedially from the ventrolateral posterior hypothalamus to the dorsal hypothalamic area. Many are oriented vertically to the thalamus in the midline. MAO-B-ir axons and fibers were detectable throughout the hypothalamus and overlapped the areas distributing HA-ir fibers. They were, however, weaker in staining intensity and apparently fewer than the HA-ir fibers. MAO-B-ir glial cells were numerous in all hypothalamic structures rich in HA-ir fibers. These results suggest that the metabolism of t-MHA may also occur within HA terminals and glial cells.

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Baroreceptor reflex-linked changes in catechol metabolism in the rat rostral ventrolateral medulla.

1. Using in vivo voltammetry, this study relates catecholamine metabolism within the rat rostral ventrolateral medulla to the level of mean arterial pressure (MAP) under halothane anaesthesia. 2. A vasopressor region was circumscribed with electrical stimulations in an area located 1000-1700 microns rostral to the obex. A catechol signal was then ascertained within this area. The recording site was surrounded with phenyl-N-methyl-ethanolamine transferase immuno-positive cell bodies. 3. Three levels of decrease of arterial pressure were induced with nitroprusside infusion: -15, -35 and -55 mmHg (n = 5 in each group) from baseline for 30 min. This led to increases in the catechol signal which were inversely related to the degree of hypotension (P < 10(-4) vs. saline for the 35 and 55 mmHg groups, P < 0.05 for the 35 mmHg group as compared to the 15 and 55 mmHg groups following recovery from hypotension). 4. Following sino-aortic deafferentation, nitroprusside-induced hypotension (-35 mmHg) did not lead to any change in the catechol signal in the rostral ventrolateral medulla (n = 5). Furthermore, controlled hypotension induced in intact rats did not evoke any change in the catechol signal recorded in a dopaminergic area of the midbrain, the ventral tegmental area (A10 area; n = 5). 5. An infusion of phenylephrine increased MAP by 35 mmHg from a baseline pressure of 105 mmHg for 30 min and evoked a non-significant decrease in the catechol signal (n = 5). In another group of rats a lower baseline pressure (80 mmHg) was stabilized (n = 5) with a higher concentration of halothane. An identical increment in pressure was then produced by a phenylephrine infusion and led to a significant reduction in the catechol signal (P < 0.05 vs. saline under similar conditions; n = 5). 6. The new findings of this study are that the level of activity of the metabolism of catecholamine in the rostral ventrolateral medulla (i) is continuously related to the level of arterial pressure, (ii) functions close to its resting level under baseline conditions and is primarily engaged during hypotension and (iii) is baroreflex linked. 7. Given the lack of direct evidence for a link between unit activity and catechol metabolism, these changes in catechol activity, recorded continuously in vivo next to adrenergic cell bodies, may represent the biochemical-specific counterpart of changes in the level of electrical unitary activity of presumed adrenergic cardiovascular medullospinal sympathoexcitatory neurons. Therefore, it provides evidence that adrenaline-synthesizing neurons in the rostral ventrolateral medulla respond to baroreceptor inputs.

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Anatomical distribution of prolactin-like immunoreactivity in the rat brain.

The present study examines prolactin PRL-like immunoreactivity (PRL-LIR) in the rat central nervous system and describes the distribution of labeled perikarya and fibers using a specific antiserum to ovine PRL. This antiserum does not cross-react with molecules of the pro-opiomelanocortin (POMC) family and recognizes rat PRL. PRL-LIR cell bodies are found exclusively in the lateral hypothalamic area surrounding the fornix, especially dorsolateral to it. No labeled cells are detectable in any other part of the brain, including the arcuate nucleus. Labeled fibers are dispersed in almost all parts of the brain. Dense plexuses are observed in the hypothalamus, midline thalamus nuclei, bed nucleus of stria terminalis, raphe dorsalis, and locus coeruleus. There is no apparent decrease in the number of PRL-LIR cell bodies and fibers in hypoprolactinemic mutant rats or after hypophysectomy, suggesting that central PRL is synthesized in such hypothalamic neurons. Comparison of PRL and alpha-melanocyte-stimulating hormone immunostainings provides evidence that the PRL network is independent of those of POMC and melanin-concentrating hormone. The present results support the hypothesis of two independent PRL systems: one peripheral (pituitary gland) and the other cerebral. Concerning the functional role of brain PRL, its widespread projections suggest that PRL is involved in multiple regulations. The presence of PRL-LIR in brain areas involved in sleep-wake control is a strong argument for its role in such a regulation.

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Dopaminergic neurons in the cat dorsal motor nucleus of the vagus, demonstrated by dopamine, AADC and TH immunohistochemistry.

In the rostral part of the dorsal motor nucleus of the vagus of the cat, neurons do not contain histochemically detectable catecholamines, even though many perikarya contain both intense aromatic L-amino acid decarboxylase (AADC) immunoreactivity and strong monoamine oxidase enzymatic activity. Similarly located perikarya have distinct immunoreactivities to tyrosine hydroxylase (TH) and dopamine after treatment with colchicine. Since inhibition of monoamine oxidase fails to reveal dopamine in these cells, its absence in non-colchicine-treated animals cannot be due to rapid deamination. It appears that dopamine is synthesized by TH and AADC in dorsal motor vagal cells and is then rapidly transported from the perikarya.

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Distribution of enkephalin and its relation to serotonin in cat and monkey spinal cord and brain stem.

The distribution of enkephalin (ENK)-like immunoreactivity (LI) in spinal cord and medulla oblongata of cat and gray monkey (Macaca fascicularis) was studied by use of immunofluorescence and peroxidase antiperoxidase (PAP) techniques. Possible coexistence between ENK- and 5-hydroxytryptamine (5-HT)-LI was also analyzed with double labeling immunofluorescence. Furthermore, in situ hybridization was used to demonstrate cell bodies in the brain stem expressing mRNA encoding for ENK. ENK-immunoreactive (IR) axonal varicosities and fibers were demonstrated throughout the spinal cord gray matter, with the highest density in the superficial dorsal horn, the area around the central canal, the intermediolateral cell column, the sacral parasympathetic nucleus, and in Onuf's nucleus. In the monkey ventral horn, ENK-IR varicose fibers could in some cases be demonstrated in very close apposition to cell bodies. A low degree of co-localization between ENK- and 5-HT-LI was seen in the spinal cord of both species. Still, fibers containing both compounds could as a rule be demonstrated in every section studied. The highest degree of coexistence was encountered in the motor nucleus of the ventral horn. Six weeks after a low thoracic spinal cord transection a decreased staining for ENK-LI was demonstrated in the ventral horn motor nucleus, whereas other parts of the spinal cord appeared unaffected. In the brain stem of cats after colchicine treatment, ENK-LI was found in a majority of the 5-HT-IR cell bodies in the raphe nuclei (nucleus raphe magnus, pallidus and obscurus) and in the lateral reticular nucleus (rostroventrolateral reticular nucleus). In cat not pretreated with colchicine, a few weakly stained ENK-IR cell bodies could be found in the midline raphe nuclei and in the lateral reticular nucleus with the PAP technique. In the monkey brain stem without colchicine treatment, using the PAP technique, heavily stained ENK-IR cell bodies could be seen in the lateral reticular nucleus whereas, as in the cat, only a few, weakly stained ENK-IR cell bodies could be seen in the midline raphe nuclei. Using in situ hybridization technique, ENK mRNA expressing cells were demonstrated in the lateral reticular nucleus while no convincing mRNA signal could be found over cell bodies in the raphe nuclei. It is concluded that part of the ENKergic innervation of the cord in both species derives from supraspinal or suprasegmental levels.(ABSTRACT TRUNCATED AT 400 WORDS)

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Combined use of silver staining of the retrograde tracer WGAapoHRP-Au and pre-embedding immunocytochemistry for electron microscopy: demonstration of dopaminergic terminals in synaptic contact with striatal neurons projecting to the substantia nigra in the rat.

We investigated the applicability of the pre-embedding immunoperoxidase technique to WGAapoHRP-Au retrograde tracing. After injection of the tracer into the substantia nigra of rat, the brain was fixed and cryostat sections were immunostained for dopamine. The sections were osmicated and silver-stained to amplify the colloidal gold particles. Products of both the immunoperoxidase staining and the silver staining could be detected and distinguished by electron microscopy at low magnification. The ultrastructure was so well preserved that synaptic characteristics could be investigated. Dopaminergic terminals were demonstrated to synapse with striatal neurons projecting to the substantia nigra.

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Immunohistochemical evidence for the presence of type B monoamine oxidase in histamine-containing neurons in the posterior hypothalamus of cats.

Using a double immunostaining method, we demonstrated that type B monoamine oxidase (MAO-B) immunoreactivity was present in virtually all histamine (HA)-immunoreactive neurons in the posterior hypothalamus of the cat. Not all MAO-B-positive neurons, however, displayed HA immunoreactivity: a minor group of neurons immunoreactive for MAO-B alone was observed in the area dorsolateral to the caudal arcuate nucleus. The results suggest that the degradation of tele-methylhistamine might occur within the intraneuronal structures of histaminergic neurons.

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Neurotoxic lesion of the mesencephalic reticular formation and/or the posterior hypothalamus does not alter waking in the cat.

In order to re-evaluate the role of two putative waking systems, we injected a neural cell body toxin, ibotenic acid (IA) (45 micrograms/microliters), into the mesencephalic reticular formation (MRF) and/or the posterior hypothalamus (PH). On the one hand, when the cell body destruction was only restricted to the MRF, the IA microinjection was followed by a temporary high voltage and slow neocortical electroencephalogram (EEG) during the first 24 postoperative hours and by a subsequent long term increase in waking which lasted 8-12 h. After the first postoperative day, there were no motor disturbances, no aphagia nor adypsia, no alteration of cortical activation and no modification of thermoregulation or of the sleep-waking cycle. On the other hand, the IA microinjection into the PH induced a hypothermia during the first postoperative night and a dramatic transient hypersomnia immediately after the disappearance of the anesthesia (14-24 h after the IA injection). On the third day, all cats recovered control level of paradoxical sleep (PS), slow wave sleep (SWS) and cerebral temperature. They presented normal motor behavior but they were not able to eat by themselves during the first postoperative week. Finally, when the lesions of the MRF and the PH were realized in one single operation, the cats were first motionless in a comatose state for 2-3 days. This state was accompanied by a transitory hypothermia and the suppression of a spontaneous or evoked cortical low voltage fast activity. However, from the 2nd postoperative week, both behavioral and EEG waking re-occurred. By contrast, the two successive operations (MRF followed by PH) did not induce a comatose state. We did not observe any deficit in motor behavior, and the sleep-waking cycle was quite normal as from the second postlesion day. In the MRF-PH-lesioned cats, the injection of alpha-methyl-p-tyrosine (150 mg/kg) induced a large decrease in waking and a moderate increase in PS. In the MRF-lesioned cats, IA produced a large area of cell body loss, centered in the MRF, that extended from levels A2 to A6 of stereotaxic planes and sometimes encroached upon the red nucleus and the substantia nigra. In the PH-lesioned cats, the histological analysis revealed a great loss of cell bodies in the PH extended from levels A8 to A12.5. The damage included the lateral and posterior hypothalamic areas and sometimes the tuberomamillary nucleus. In MRF- and PH-lesioned cats, the cell body loss extended from levels A2 to A12.5.(ABSTRACT TRUNCATED AT 400 WORDS)

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Distribution of type B monoamine oxidase immunoreactivity in the cat brain with reference to enzyme histochemistry.

We studied the detailed distributions and morphology of structures immunoreactive to type B monoamine oxidase, and compared them with those stained by monoamine oxidase enzyme histochemistry in the brain of cats treated with or without colchicine. By means of the indirect immunohistochemical method in conjunction with type B monoamine oxidase monoclonal antibody, we demonstrated type B monoamine oxidase immunoreactivity in neuronal cell bodies, fibers and astroglial cells in the cat brain. As expected, the distribution of type B monoamine oxidase-immunoreactive cell bodies overlapped that of serotonin-containing ones in the lower brainstem and midbrain, as well as that of histaminergic ones in the posterior hypothalamus. We found novel cell groups containing type B monoamine oxidase in the areas described below. Intense type B monoamine oxidase-immunopositive and enzymatically active neurons, corresponding to liquor-contact ones, were discovered in the wall of the central canal of the spinomedullary junction. Weak immunoreactivity was identified in neurons of the dorsal motor nucleus of the vagus, parvocellular reticular formation and locus coeruleus complex, which have been reported to contain type A monoamine oxidase enzymatic activity. Type B monoamine oxidase-immunostaining in these structures was enhanced by treatment with colchicine. In addition, lightly immunostained cells were distinguished in the caudal portion of the hypothalamic arcuate nucleus, area of tuber cinereum, retrochiasmatic area, and rostral portion of the paraventricular thalamic nucleus after colchicine treatment. These cells also displayed monoamine oxidase activity; however, it was difficult to enzymatically characterize their nature due to its weak activity and sensitivity to inhibitors of both A and B. Distinct type B monoamine oxidase-immunoreactive fibers and terminal-like dots were abundant in the whole brain, particularly in the central gray, dorsal pontine tegmentum, interpeduncular and pontine nuclei, nucleus of the solitary tract and dorsal motor nucleus of vagus, where dense innervations of serotonergic fibers have been reported. Their immunoreactive density increased after colchicine treatment, but monoamine oxidase enzymatic reaction did not. An intense immunoreactivity could be seen in many glial cells in parts of the brain including myelinated axon pathways. The densest accumulation of such labeled glial cells was found in the central gray, inferior olive, medial geniculate body, substantia nigra, ventral tegmental area of Tsai, retrorubral area, hypothalamus, thalamus and bed nucleus of the stria terminalis. In contrast, the striatum contained less numerous type B monoamine oxidase-immunoreactive and enzymatically active astroglial cells in comparison with the other structures.(ABSTRACT TRUNCATED AT 400 WORDS)

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Monoaminergic interaction in the central nervous system: a morphological analysis in the locus coeruleus of the rat.

The locus coeruleus of the rat is richly innervated by many aminergic neurons varying in amine content and in site of origin. There are adrenergic and noradrenergic neurons originating in the medulla oblongata, dopaminergic from the hypothalamus, serotonergic from the mesencephalon and also intrinsic noradrenergic neurons in the locus coeruleus complex. Of these, adrenergic and dopaminergic inputs appear relatively specific and powerful.

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Regulation of neurotransmitter synthesis: from neuron to gene.

We are interested in the molecular mechanisms of the regulation of neurotransmitter related gene expression by neurotrophic factors and neuronal activity. Previous work has shown that conditioned medium of muscle cells induces choline acetyltransferase (CAT) activity and represses tyrosine hydroxylase, dopamine-beta-hydroxylase and aromatic L-amino acid decarboxylase (AADC) activities in cultured sympathetic neurons. Here, we show that a new muscle-derived cell line secretes two factors which induce CAT activity in spinal cord cultures; one of those is related to LIF, a CAT inducing factor for sympathetic neurons. Preliminary data are presented on the structure of the human AADC and CAT genes. Putative promoter regions have been coupled to reporter genes; transient transfection experiments will allow us to determine the promoter elements responsible for the regulation by neurotrophic factors. We also summarize the distribution of AADC-immunoreactive cells in rat and cat brain, which will be used as a reference for the study of the specificity of expression of AADC promoter in transgenic mice.

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Aromatic L-amino acid decarboxylase-immunohistochemistry in the cat lower brainstem and midbrain.

By indirect immunohistochemistry, the present study examined the distribution of neuronal structures in the cat medulla oblongata, pons, and midbrain, showing immunoreactivity to aromatic L-amino acid decarboxylase (AADC), which catalyzes the conversion of L-3, 4-dihydroxyphenylalanine (L-DOPA) to dopamine, and 5-hydroxytryptophan to serotonin (5HT). With simultaneous and serial double immunostaining techniques, immunoreactivity to this enzyme was demonstrated in most of the catecholaminergic and serotonergic neurons. We could also demonstrate AADC-IR cell bodies that do not contain tyrosine hydroxylase (TH-) or 5HT-immunoreactivity (called "D-type cells") outside such monoaminergic cell systems. At the medullo-spinal junction, very small D-type cells were found within and beneath the ependymal layer of the 10th area of Rexed surrounding the central canal. D-type cells were localized in the caudal reticular formation, nucleus of the solitary tract, a dorsal aspect of the lateral parabrachial nucleus, and pretectal areas as have been reported in the rat. Furthermore, the present study describes, in the cat brainstem, new additional D-type cell groups that have not been reported in the rat. Dense or loose clusters of D-type cells were localized in the external edge of the laminar trigeminal nucleus, dorsal motor nucleus of the vagus, external cuneate nucleus, nucleus praepositus hypoglossi, central, pontine, and periaqueductal gray, superficial layer of the superior colliculus, and area medial to the retroflexus. D-type cells were loosely clustered in the lateral part of the central tegmental field dorsal to the substantia nigra, extending dorsally in the medial division of the posterior complex of the thalamus and medial side of the brachium of the inferior colliculus. They extended farther rostrodorsally along the medial side of the nucleus limitans and joined with the pretectal cell group. Almost all these cells were very small and ovoid to round with 1-2 short processes with the exception of dorsal motor vagal cells. AADC-IR axons were clearly identified in the vagal efferent nerves, longitudinal medullary pathway, dorsal tegmental bundle rostral to the locus coeruleus. Serotonergic axons were identified not only in the central tegmentum field and lateral side of the central superior nucleus, but also in the ventral surface of the medulla oblongata. We describe principal densely stained fiber plexuses in the cat brainstem. The findings of the present study provide a morphological basis for neurons that decarboxylate endogenous and exogenous L-DOPA, 5HTP, and other aromatic L-amino acids.

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Dopamine- and dopa-immunoreactive neurons in the cat forebrain with reference to tyrosine hydroxylase-immunohistochemistry.

The distribution of cell bodies containing immunoreactivities to dopamine (DA), L-3,4-dihydroxyphenylalanine (DOPA) and tyrosine hydroxylase (TH) was studied immunohistochemically in the cat forebrain especially in the hypothalamus with or without intraventricular administration of colchicine. In normal cats, DA-immunoreactive (IR) neurons, whose intensity of immunostainings was variable from one to another, were localized exclusively in the hypothalamus and showed a distribution pattern similar to that of TH-IR ones. They were distributed in the posterior, dorsal and periventricular hypothalamic areas. Arcuate cells showed no or very weak DA-immunoreactivity. Weak to intense DOPA-IR cells were distributed in a similar manner to DA-IR ones but were far smaller in number. In colchicine-treated animals, DA- and DOPA-immunoreactivities were enhanced particularly in arcuate and dorsal hypothalamic cells. A cluster composed of small DA- and DOPA-IR cells was identified in the area ventral to the mamillothalamic tract equivalent to rat A13c TH-IR cell group. Colchicine treatment enabled us to visualize a large number of TH-IR perikarya in the medial and lateral preoptic areas, anterior commissure nucleus, basal forebrain, area closely related to the organum vasculosum laminae terminalis, and some in the bed nucleus of the stria terminalis as has been reported in other species. However, virtually none of these cells contained detectable DA- and DOPA-immunoreactivities.

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Neurons containing messenger RNA encoding glutamate decarboxylase in rat hypothalamus demonstrated by in situ hybridization, with special emphasis on cell groups in medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus.

Previous deafferentation studies have suggested that most hypothalamic GABAergic innervation originates from neurons within the hypothalamus. We have investigated the distribution of GABAergic cell groups in the rat hypothalamus by means of the in situ hybridization technique, using a cDNA probe for messenger RNA encoding glutamate decarboxylase. Several major GABAergic cell groups were demonstrated, including cells of the tuberomammillary nucleus, arcuate nucleus, suprachiasmatic nucleus, medial preoptic area, anterior hypothalamic area, the dorsomedial hypothalamic nucleus, perifornical area, and lateral hypothalamic area. The most prominent glutamate decarboxylase mRNA-containing cell groups were located in the medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus, and were composed of small- to medium-sized neurons. Compared to previously well-characterized GABAergic cell groups in the tuberomammillary nucleus, reticular thalamic nucleus, and non-pyramidal cells of cerebral cortex, the cells of these GABAergic groups demonstrated only weak cDNA labelling, indicating that they contain lower levels of glutamate decarboxylase mRNA. Several types of control experiments supported the specificity of this cDNA labelling, and the GABAergic nature of these cell populations was further supported by detection of glutamate decarboxylase and GABA immunoreactivity. Abundance of GABAergic cells in many hypothalamic nuclei indicates that GABA represents quantitatively the most important transmitter of hypothalamic neurons, and may be involved in neuroendocrine and autonomic regulatory functions.

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