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K Kitahama

Publications and source records attributed to K Kitahama.

At least 73 records · Page 4Linked to original sources

Adrenergic input from medullary ventrolateral C1 cells to the nucleus raphe pallidus of the cat, as demonstrated by a double immunostaining technique.

By means of a double immunostaining technique using unconjugated cholera-toxin B subunit (CTb) as a retrograde tracer combined with phenylethanolamine-N-methyltransferase (PNMT) immunohistochemistry, we demonstrated that the nucleus raphe pallidus of the cat receives a major projection from the ventrolateral part of the rostral medulla corresponding to the nucleus paragigantocellularis lateralis and the ventrolateral medullary reticular formation just caudal to it. We further showed that nearly 60% of the total CTb-labeled cells in this region are immunoreactive to PNMT. These double-labeled cells constitute one-third of the total PNMT-immunoreactive cells.

Adrenergic Fibers↗

Demonstration of GABAergic cell bodies in the suprachiasmatic nucleus: in situ hybridization of glutamic acid decarboxylase (GAD) mRNA and immunocytochemistry of GAD and GABA.

The existence of GABAergic neurons in the rat suprachiasmatic nucleus (SCN) was demonstrated by three specific markers; mRNA coding for glutamic acid decarboxylase (GAD) and visualized by in situ hybridization using a 35S-labelled cDNA probe, and GAD protein and GABA were identified by immunocytochemistry using specific antisera. In situ hybridization demonstrated well labelled GAD mRNA positive cells throughout SCN, and GABA and GAD immunoreactive cells showed similar distributions. These results indicate that GABA is a transmitter of a large portion of the SCN neuronal population.

Animals↗

Type B monoamine-oxidase-containing cells and fibers in the cat hypothalamus demonstrated by an improved enzyme histochemical method.

The present study, using a diaminobenzidine (DAB)-coupled peroxidation method, examined the distribution and morphological characteristics of neuronal structures containing type B monoamine oxidase (MAO-B) in the cat hypothalamus. Large and intensely stained, distinctive MAO-B-positive cells, multipolar and with long dendritic arbors, were principally distributed in the ventral hypothalamus extending from A7 to A12.5 of the Horsley-Clarke plane. These cells were located caudally in the ventral surface of the brain including the tuberomamillary nucleus (TM) and the region surrounding the mamillary nuclei. Rostrally, they were aggregated in the area surrounding the fornix, particularly in the lateral perifornical region, and dispersed in the anterior mamillary nucleus, lateral hypothalamic area (HLA), and the medial tip of the entopeduncular nucleus. The most rostral positive cell group was identified in a narrow space between the optic tract and the entopeduncular nucleus at the A12.5 level. In addition to these large cells, the present study disclosed the presence of "small" to "very small" MAO-B-positive cells in the area surrounding the mamillary recess and the lateral part of the caudal arcuate nucleus. Distinct MAO-B-stained fibers were identified in all regions of the hypothalamus. A large number of thick labeled fibers were observed in the ventral hypothalamus including the TM and premamillary nucleus and posterior and lateral hypothalamic areas. A dense network of MAO-B-positive terminal-like fibers was observed in the dorsomedial nucleus where very small labeled cells were scattered. Many intensely stained thick and straight fibers were seen running ventrolaterally in the anterior part of the HLA and in the narrow space between the entopeduncular nucleus and optic tract. In the area of the tuber cinereum and the ventral part of the HLA, there were many positive fibers cut transversely, possibly projecting to the more anterior parts of the brain such as the diagonal band of Broca or septal nuclei.

Animals↗

A new group of tyrosine hydroxylase-immunoreactive neurons in the cat thalamus.

A new cell group composed of a large number of neurons immunoreactive to tyrosine hydroxylase (TH) was demonstrated in the paraventricular nucleus and midline nuclei of the cat thalamus, using four different anti-TH sera after colchicine treatment. However, in these regions, we did not detect any cell bodies containing other catecholamine synthesizing enzymes nor dopamine.

Animals↗

Comparative marker analysis of the ependymocytes of the subcommissural organ in four different mammalian species.

The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.

Animals↗

5-Hydroxytryptophan uptake and decarboxylating neurons in the cat hypothalamus.

Parachlorophenylalanine, an inhibitor of tryptophan hydroxylase, induced a virtually total disappearance of serotonin-immunoreactivity in the hypothalamus of the cat. After intrahypothalamic injection of 5-hydroxytryptophan, an immediate precursor of serotonin in cats pretreated with parachlorophenylalanine, serotonin-immunoreactivity was detected in many fibers surrounding the injection site. Furthermore, when 5-hydroxytryptophan was injected with inhibitor of monoamine oxidase, a large number of small neurons immunoreactive to serotonin was identified in many discrete regions: the anterior and lateral hypothalamic areas, preoptic area, suprachiasmatic nucleus, dorsal hypothalamic area, dorsomedial nucleus, posterior hypothalamic area and nucleus of the fields of Forel. Serotonin-immunoreactivity was also evident in the thick axon bundles in the lateral hypothalamus. The distribution pattern of these cells was quite similar to that of aromatic L-amino acid decarboxylase, which catalyses the conversion of 5-hydroxytryptophan to serotonin and that of L-3,4-dihydroxyphenylalanine to dopamine. However, we failed to demonstrate serotonin-immunoreactivity in these parvocellular neurons without monoamine oxidase inhibitor. It is possible that 5-hydroxytryptophan is decarboxylated to serotonin by aromatic L-amino acid decarboxylase but rapidly degraded by monoamine oxidase-A, the enzyme which preferentially deaminates serotonin. In contrast, serotonin-immunostaining was always demonstrable after intrahypothalamic injection of 5-hydroxytryptophan without monoamine oxidase inhibitor in magnocellular neurons located in the ventrolateral posterior hypothalamus and which contain exclusively monoamine oxidase-B and histidine decarboxylase. It appears that in these cells and axons, serotonin, possibly formed by histidine decarboxylase, is not rapidly oxidized by monoamine oxidase-B. Possible roles of serotonin as a neurohormone in sleep-waking regulation and of trace amines in the brain are discussed.

5-Hydroxytryptophan↗

Reversibility of para-chlorophenylalanine-induced insomnia by intrahypothalamic microinjection of L-5-hydroxytryptophan.

Para-chlorophenylalanine, a blocker of serotonin biosynthesis by inhibiting tryptophan hydroxylase, induced total insomnia which was accompanied in cat by a permanent discharge of ponto-geniculo-occipital activity. L-5-Hydroxytryptophan microinjection (1-4 micrograms/0.5 microliters) in the anterior hypothalamus 72 h after para-chlorophenylalanine administration, restored both slow wave sleep and paradoxical sleep with variable latencies for each state of sleep. On the contrary, ponto-geniculo-occipital activity was never suppressed. The hypnogenic effects of L-5-hydroxytryptophan were always followed by a return of the para-chlorophenylalanine-induced insomnia. On the other hand, the temperature recording did not show any alteration of the cerebral temperature after para-chlorophenylalanine treatment but the subsequent L-5-hydroxytryptophan microinjection was followed by hyperthermia. Using immunohistochemistry for serotonin after intrahypothalamic L-5-hydroxytryptophan microinjection in parachlorophenylalanine-pretreated cat, we defined a restricted region of the anterior hypothalamus possibly responsible for the hypnogenic effect. This region included the lateral hypothalamus and the anterior hypothalamic area. It is suggested that the reversible hypersomnia after L-5-hydroxytryptophan microinjection in the anterior hypothalamus in para-chlorophenylalanine-pretreated cat is due to a neurohormonal action of serotonin: serotonin could act upon the anterior hypothalamus which secondarily inhibits a waking system located in the ventrolateral hypothalamus leading to the appearance of paradoxical sleep.

5-Hydroxytryptophan↗

Long-lasting insomnia induced by preoptic neuron lesions and its transient reversal by muscimol injection into the posterior hypothalamus in the cat.

In order to analyse the role of the anterior hypothalamus in the regulation of the sleep-waking cycle we made bilateral neuronal lesions at different levels of the anterior hypothalamus in cats, by means of microinjections of a cell-specific neurotoxin:ibotenic acid. These lesions resulted in severe insomnia in eight cats. This insomnia was characterized by a large decrease or even disappearance of paradoxical sleep and deep slow wave sleep and, to a lesser extent, by a decrease of light slow wave sleep, for 2-3 weeks. In the other five animals, we observed a large reduction of deep slow wave sleep (0-40% of control level), but a less intensive decrease of time spent in paradoxical sleep (50-75% of control level) and no marked effect on light slow wave sleep. During the first 3-6 postoperative days we also noticed hyperthermia in all cats; thereafter, the animals presented only a slight increase in brain temperature which did not appear to trigger the sleep impairment. Histological analysis of the different lesions revealed that the insomnia could be attributed to neuronal cell body destruction in the mediobasal part of the anterior hypothalamus covering; the medial preoptic area and a narrow portion of the lateral preoptic area as well as a restricted part of the anterior hypothalamic nucleus. In order to investigate the putative role of the posterior hypothalamic structures in the mechanism of insomnia after lesion of the mediobasal preoptic area neurons we injected an agonist of GABA into the ventrolateral part of the posterior hypothalamus to locally depress the neuronal activity. The bilateral intracerebral microinjection of muscimol (0.5-5 micrograms) induced a transient intensive hypersomnia (slow wave sleep and paradoxical sleep). These findings indicate that neuronal cell loss in the mediobasal preoptic area induced a long lasting insomnia. Thus, it may be hypothesized that the integrity of this structure is necessary for sleep appearance. Finally, our data are in keeping with an intrahypothalamic regulation of the sleep-waking cycle.

Animals↗

Serotonin neurons and their physiological roles.

Anatomical and functional aspects of serotonin (5HT) neurons in the central nervous system are described with special reference to their neurohumoral function. The central 5HT neurons show morphological features characteristics of the modulating nervous system: a small number of cell bodies and extremely broad innervation. These may play some part in EEG arousal through the divergent projection. Being most active during the waking state, the 5HT neurons are responsible for the onset of sleep. 5HT released during waking may initiate the humoral, as well as neuronal mechanisms leading to sleep. Although the 5HT dependent hypogenic structure is highly localized in the anterior hypothalamus, the effect seems to be due to neurohumoral action of 5HT. The neurohumoral function of the 5HT neuron is suggested by the ultrastructure of 5HT terminals almost devoid of a synaptic structure (less than 10%). Intraventricular 5HT fibers also represent a substrate supporting the neurohumoral function. A strong activity of monoamine oxidase (MAO) is found in intercellular spaces in the arcuate nucleus region of the hypothalamus, where a high 5HT immunoreactivity is seen in the young rat before MAO activity appears. It is likely that 5HT liberated in the cerebrospinal fluid is drained through these intercellular channels.

Animals↗

[Cells presenting GABA immunoreactivity in the hypothalamus of the cat].

The distribution of GABA-immunoreactive (IR) cells was studied by immunohistochemistry in conjunction with highly specific antiserum GABA in the cat hypothalamus. Colchicine pretreatment made it possible to visualize a large number of labeled cells in the medial preoptic and dorsal hypothalamic areas. In contrast, the ventromedial and anterior hypothalamic nuclei contained only a few labeled cells, and the paraventricular and supraoptic nuclei were devoid of them. A very dense network of GABA-IR presumptive terminals was seen in the ventrolateral posterior hypothalamus where labeled cells could be recognized. The possibility of an involvement of the GABAergic neuronal system in the regulation of sleep-waking cycle is discussed.

Animals↗

[Prolactin-secreting neurons in the dorsolateral hypothalamus in Sprague-Dawley rats].

By means of immunocytochemical techniques ovine prolactin like immunoreactivity (oPRL-LIR) has been demonstrated in the perikarya located around fornix in the dorso-lateral part of the rat hypothalamus. No PRL-LIR was observed in the arcuate n. perikarya. Immunoreactive fibers were present in the hypothalamus, medial thalamus, accumbens and amygdaloid nuclei.

Amygdala↗

L-dopa-immunoreactive neurons in the rat hypothalamic tuberal region.

The presence of L-DOPA-immunoreactivity is reported for the first time in the rat hypothalamic tuberal region. L-DOPA-immunoreactive neurons were demonstrated to be present in the ventrolateral part of the arcuate nucleus and periarcuate region just dorsal to the ventral surface of the brain (VLAR/PA). Weakly L-DOPA-immunostained neurons were found in the dorsomedial part of the arcuate nucleus and its neighboring periventricular nucleus (DMAR/PV). In contrast, dopamine (DA)-immunoreactive neurons were detected only in the DMAR/PV. These findings suggest that L-DOPA exists not only as a precursor of DA in neurons of the DMAR/PV, but also as an end-product in cells of the VLAR/PA.

Animals↗

Endogenous L-dopa, its immunoreactivity in neurons of midbrain and its projection fields in the cat.

L-DOPA (L-3,4-dihydroxyphenylalanine) immunoreactivity was demonstrated in neurons of the cat ventral midbrain and its projection areas, using an immunohistochemical method in conjunction with a newly developed highly specific anti-L-DOPA serum. L-DOPA-immunoreactive (IR) neurons were found in the substantia nigra, retrorubral area and ventral tegmental area of Tsai. L-DOPA-labeled fibers and terminals were hardly detectable in the nigrostriatal pathway and in the caudate nucleus which showed very intense dopamine-immunoreactivity. In contrast, many short labeled processes were detectable in the central amygdala and, although very few in number, in the entorhinal cortex.

Animals↗

Comparative topography of dopamine- and tyrosine hydroxylase-immunoreactive neurons in the rat arcuate nucleus.

The distribution of dopamine (DA)-immunoreactive (IR) cells is described in the rat arcuate nucleus of the hypothalamus and its adjacent areas and compared with that of tyrosine hydroxylase (TH)-IR cells. Small DA-IR cells were seen to be aggregated mainly in the dorsomedial part of the nucleus, but were hardly detectable in its ventrolateral portion and neighbouring periarcuate region which showed many larger TH-IR cells. This study reveals, for the first time, the differences in the respective topography of those neurons which actually contain detectable DA and those which contain TH, the initial synthesizing enzyme of catecholamine.

Animals↗

Hypophysectomy does not disturb the sleep-waking cycle in the cat.

The ablation of the hypophysis does not disturb the basic waking-sleep cycle. Further, this intervention fails to modify the recovery of sleep after instrumental paradoxical sleep deprivation as well as after injection of 5-hydroxytryptophan in parachlorophenylalanine-pretreated insomniac cats. These results demonstrate that the hypophysis does not play a significant role in sleep mechanisms. We discuss these data in view of a possible regulation of the sleep-waking cycle by hypothalamo-hypophyseal hormones.

Animals↗

Immunohistochemistry of aromatic L-amino acid decarboxylase in the cat forebrain.

The topographic distribution of aromatic L-amino acid decarboxylase (AADC)-immunoreactive (IR) neurons was investigated in the cat hypothalamus, limbic areas, and thalamus by using specific antiserum raised against porcine kidney AADC. The perikarya and main axons were mapped on an atlas in ten cross-sectional drawings from A8 to A16 of the Horsley Clarke stereotaxic plane. AADC-IR neurons were widely distributed in the anterior brain. They were identified in the posterior hypothalamic area, rostral arcuate nucleus of the hypothalamus, dorsal hypothalamic area, and periventricular complex of the hypothalamus, which contain tyrosine hydroxylase (TH)-IR cells and are known as A11 to A14 dopaminergic cell groups. AADC-IR perikarya were also found in the other hypothalamic areas where few or no TH-IR cells have been reported: the supramamillary nucleus, tuberomamillary nucleus, pre- and anterior mamillary nuclei, caudal arcuate nucleus, dorsal hypothalamic area immediately ventral to the mamillothalamic tract, anterior hypothalamic area, area of the tuber cinereum, retrochiasmatic area, preoptic area, suprachiasmatic and dorsal chiasmatic nuclei. We also identified them in the anterior commissure nucleus, bed nucleus of the stria terminalis, stria terminalis, medial and central amygdaloid nuclei, lateral septal nucleus, and nucleus of the diagonal band of Broca. AADC-IR neurons were localized in the ventromedial part of the thalamus, lateral posterior complex, paracentral nucleus and lateral dorsal nucleus of the thalamus, medial habenula, parafascicular nucleus, subparafascicular nucleus, and periaqueductal gray. Conversely, we detected only a few AADC-IR cells in the supraoptic nucleus whose rostral portion contains TH-IR perikarya. Comments are made on the relative localizations of the AADC-IR and TH-IR neurons, on species differences between the cat and rat, as well as on the possible physiological functions of the enzyme AADC.

Animals↗

Topography of neurophysin-immunoreactive neurons projecting to the neurohypophysis: direct evidence as revealed by a double staining method.

The topographic distribution of neurophysin-immunoreactive (NP-IR) cells projecting to the posterior pituitary gland has been studied in the cat using a double staining method: immunohistochemistry of neurophysin in conjunction with horseradish peroxidase (HRP) retrograde tracer technique. We found that almost all the hypothalamic NP-IR cells project directly to the neurohypophysis except those localized in the suprachiasmatic nucleus and in the caudal part of the paraventricular nucleus.

Animals↗

Localization of CRF-immunoreactive neurons in the cat medulla oblongata: their presence in the inferior olive.

Corticotropin releasing factor (CRF)-immunoreactive (IR) perikarya, visualized by the indirect immunoperoxidase method in colchicine-pretreated cats, were localized in many discrete regions of the medulla oblongata. They were found mainly in the dorsal aspect and midline of the medulla oblongata, and more rostrally in the ventrolateral portion. Our results also demonstrated CRF-IR neurons in the rostrocaudal extent of the inferior olive, probably projecting to the cerebellar cortex via thick axons visualized along the lateral edge of the medulla. CRF-IR olivary cells were also found in the pontine cat from which the forebrain was removed, but neither in hypophysectomized nor adrenalectomized cats.

Animals↗