Biomedical subjects

N Mizuno

Publications and source records attributed to N Mizuno.

At least 199 records · Page 11Linked to original sources

PubMed1993

Serotoninergic projections from the dorsal raphe nucleus to the nucleus submedius in the rat and cat.

The nucleus submedius in the medial thalamus has been known to receive spinothalamic and trigeminothalamic fibers, and to contain neurons which can be activated by noxious stimuli. These previous findings suggest that the nucleus submedius may be involved in the processing and relay of pain-related information. In the present study, we immunohistochemically observed in the rat and cat that the nucleus submedius was distributed with a considerable amount of serotoninergic fibers. After iontophoretic injection of cholera toxin B subunit into the nucleus submedius, the sequential double-antigen immunofluorescence histochemistry for retrogradely transported cholera toxin B subunit and serotonin revealed that the serotoninergic fibers to the nucleus submedius arose mainly from the dorsal raphe nucleus, and additionally from the ventrolateral and medial parts of the midbrain periaqueductal gray. The direct projections from the dorsal raphe nucleus to the nucleus submedius were confirmed by anterograde axonal tracing after iontophoretic injection of Phaseolus vulgaris-leucoagglutinin into the dorsal raphe nucleus. The disappearance of almost all serotoninergic fibers in the nucleus submedius was also observed after destruction of the dorsal raphe nucleus. The fluorescent retrograde double-labeling with Diamidino Yellow and Fast Blue further revealed that some neurons in the dorsal raphe nucleus projecting directly to the nucleus submedius sent their axon collaterals to the ventrolateral orbital region of the cerebral cortex, nucleus accumbens, amygdala, nucleus raphe magnus, caudal spinal trigeminal nucleus, or spinal cord. The possible roles of the serotoninergic projections from the dorsal raphe nucleus to the nucleus submedius in pain control and/or the olfactolimbic functions are discussed.

Animals↗

PubMed1993

Effects of ultraviolet-B and PUVA on ornithine decarboxylase activity, DNA synthesis, and protein kinase C activity in mouse skin.

Ultraviolet-B and PUVA share several biological events with phorbol ester tumor promoters. The effects of ultraviolet-B irradiation and topical PUVA treatment on ornithine decarboxylase activity, DNA synthesis, and protein kinase C activity, which are known to be induced or activated by phorbol ester tumor promoter, were investigated in hairless mouse skin. Ornithine decarboxylase activity was remarkably enhanced by ultraviolet-B and PUVA. Although PUVA did not affect DNA synthesis significantly, ultraviolet-B stimulated epidermal DNA synthesis approximately 5-fold over control values at 48 h. However, unexpectedly, neither cytosolic nor membrane-bound protein kinase C activity showed any change during the 2 h after either treatment. These results suggest that the protein kinase C system is not involved in the initial signal transduction system of ultraviolet-B or PUVA, unlike the case with phorbol ester tumor promoter.

Animals↗

PubMed1993

Effects of colchicine on the induction of ornithine decarboxylase and its gene expression by the phorbol ester tumour promoter.

The activity and gene expression of ornithine decarboxylase (ODC, an indicator of tumour promotion) were induced by the phorbol ester tumour promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), in mouse skin. In the present study, the effect of colchicine, a microtubule-disrupting agent, on ODC activity and its gene expression were investigated. On administration of colchicine (100 micrograms) intraperitoneally 1.5 h before TPA treatment, ODC activity and ODC mRNA levels stimulated by TPA were suppressed to about 52 and 64%, respectively. These results suggest the involvement of a microtubule or colchicine-sensitive substrate in the signal transduction system for gene expression.

Animals↗

PubMed1993

Percutaneous permeation of basic compounds through shed snake skin as a model membrane.

Relationships between the in-vitro permeability of basic compounds through shed snake skin as a suitable model membrane for human stratum corneum and their physiochemical properties were investigated. Compounds with low pKa values were selected to compare the permeabilities of non-ionized forms of the compounds. Steady-state penetration was achieved immediately without a lag time for all compounds. Flux rate and permeability coefficient were calculated from the steady-state penetration data and relationships between these parameters and the physicochemical properties were investigated. The results showed that permeability may be controlled by the lipophilicity and the molecular size of the compounds. Equations were developed to predict the permeability from the molecular weights and the partition coefficients of basic compounds.

Animals↗

PubMed1993

Plasma and cerebroventricular fluid levels of pancreatic polypeptide in the dog: effects of feeding, insulin-induced hypoglycemia, and physical exercise.

We investigated the penetration of plasma pancreatic polypeptide (PP) into the third cerebral ventricular fluid (CSF) of dogs. Plasma and CSF levels of PP were measured by RIA during the iv infusion of PP and during such stimuli as eating, insulin-induced hypoglycemia, and physical exercise. Plasma and CSF levels of insulin and glucose were also measured and compared during eating and insulin-induced hypoglycemia. Plasma glucose increased after feeding and decreased after insulin injection, followed by a corresponding change in CSF glucose without an apparent time lag. CSF insulin insignificantly increased after feeding and the injection of insulin, while CSF PP did not increase despite the marked elevation of plasma PP in response to these stimuli. CSF PP did not increase after the infusion of exogenous PP. Strenuous exercise, however, evoked an increase in both plasma and CSF PP levels; the CSF response was prompt, but more prolonged than that of plasma, suggesting the slow removal of PP from CSF. We conclude that 1) PP and insulin in CSF do not appear to play a major role in the short term regulation of food intake and acute changes in energy metabolism; and 2) PP, probably after entering the brain, may modulate brain function in such physiological situations as strenuous exercise.

Animals↗

PubMed1993

Effect of a high-fat meal on the bioavailability of phenytoin in a commercial powder with a large particle size.

The effect of a high-fat meal on the bioavailability of free acid phenytoin (DPH) from Hydantol powder with a large particle size (mean particle size, 190 microns) was investigated in four healthy male subjects. The drug was administered as a single oral 5 mg/kg dose of free acid DPH in the fasting state, with a low-fat meal, or with a high-fat meal using a crossover study design. Seven blood samples were collected over a 34-h period following drug administration, and the drug plasma concentrations were determined by GLC. In comparison with the fasting state results, the mean area under the plasma concentration-time curve up to infinity after administration (AUC0-infinity) and the peak plasma concentration (Cmax) of DPH from Hydantol powder significantly increased about 2-fold with the intake of the high-fat meal and about 1.5-fold with the intake of the low-fat meal. The elimination rate constant was not significantly different among the three treatments. The increased bioavailability with the high-fat meal probably resulted from accelerated dissolution of the poorly soluble Hydantol powder due to the stimulation of bile flow or delay of the gastric emptying time caused by the fat intake.

Adult↗

PubMed1993

Signal transduction, pharmacological properties, and expression patterns of two rat metabotropic glutamate receptors, mGluR3 and mGluR4.

The metabotropic glutamate receptors are coupled to intracellular signal transduction via G-proteins and consist of a family of at least five different subtypes, termed mGluR1-mGluR5. We studied the signal transduction mechanism and pharmacological characteristics of the rat mGluR3 and mGluR4 subtypes in Chinese hamster ovary cells permanently expressing the cloned receptors. Both mGluR3 and mGluR4 inhibit the forskolin-stimulated accumulation of intracellular cAMP formation in response to agonist interaction. Consistent with the high degree of sequence similarity to mGluR2, mGluR3 closely resembles mGluR2 in its agonist selectivity; the potency rank order of agonists is L-glutamate > trans-1-aminocyclopentane-1,3-dicarboxylate > ibotenate > quisqualate. mGluR4 is totally different in its agonist specificity from any other member of the metabotropic receptors. This receptor potently reacts with L-2-amino-4-phosphonobutyrate (L-AP4) in a stereo-selective manner and moderately responds to L-serine-O-phosphate. mGluR4 thus corresponds well to the putative L-AP4 receptor characterized from brain preparations. Blot and in situ hybridization analyses indicated that both mRNAs are widely distributed in the rat brain. mGluR3 mRNA is highly expressed in neuronal cells of the cerebral cortex and the caudate-putamen, and in granule cells of the hippocampal dentate gyrus. The expression pattern of mGluR4 mRNA is more restricted, and this expression is prominent in the cerebellum, olfactory bulb, and thalamus. Furthermore, the mGluR3 mRNA, unlike the other mRNAs for the metabotropic receptors, is highly expressed in glial cells throughout the brain regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Single retinal ganglion cells sending axon collaterals to the bilateral superior colliculi: a fluorescent retrograde double-labeling study in the Japanese monkey (Macaca fuscata).

Single retinal ganglion cells projecting bilaterally to the superior colliculi (SC) by way of axon collaterals were revealed in the Japanese monkey (Macaca fuscata). After injecting Fast blue into the SC on one side and Diamidino yellow into the SC on the opposite side, some retinal ganglion cells were double-labeled with both tracers. Most of them were large cells (more than 25 microns in diameter), and were localized in a narrow strip around the vertical meridian of the retina on each side. This retinal area roughly corresponds to the reported strip of nasotemporal overlap, where both crossed and uncrossed retinofugal projections arise.

Amidines↗

PubMed1992

Trigeminal ganglion neurons which project by way of axon collaterals to both the caudal spinal trigeminal and the principal sensory trigeminal nuclei.

Employing a combination of fluorescent retrograde double labeling and immunofluorescence histochemistry, we found that some single neurons in the trigeminal ganglion of the rat projected by way of axon collaterals both to the caudal spinal trigeminal nucleus and to the principal sensory trigeminal nucleus, and that about 40% or 57% of these neurons showed respectively substance P- or calcitonin gene-related peptide-like immunoreactivity.

Animals↗

PubMed1992

Expression of the mRNA for the rat NMDA receptor (NMDAR1) in the sensory and autonomic ganglion neurons.

The distribution of NMDA receptor (NMDAR1) on neurons in the peripheral ganglia was examined in the adult rat by in situ hybridization. NMDAR1 mRNA was expressed in all neurons in the sensory and autonomic ganglia examined; in the dorsal root, trigeminal, nodose, superior cervical, and sphenopalatine ganglia. Possible roles of the NMDA receptor on the sensory and autonomic ganglion neurons are discussed.

Animals↗

PubMed1992

Paucity of glutaminase-immunoreactive nonpyramidal neurons in the rat cerebral cortex.

Glutaminase has been considered to be a synthesizing enzyme of transmitter glutamate in pyramidal neurons of the cerebral cortex. In the present study, an attempt was made to examine with a double immunofluorescence method whether or not nonpyramidal neurons of the cerebral cortex are immunoreactive for glutaminase. Glutaminase was stained with mouse anti-glutaminase IgM and FITC-labeled anti-[mouse IgM] antibody. In the same section, parvalbumin (PA), calbindin (CB), choline acetyltransferase (CAT), vasoactive intestinal polypeptide (VIP), corticotropin releasing factor (CRF), cholecystokinin (CCK), somatostatin (SS), or neuropeptide Y (NPY) was visualized as a marker for nonpyramidal neurons with an antibody to each substance, biotinylated secondary antibody and Texas Red-labeled avidin. Virtually no glutaminase immunoreactivity was seen in PA-, CB-, CAT-, VIP-, CRF-, CCK-, SS-, or NPY-immunoreactive neuronal perikarya in the neocortex and mesocortex (cingulate and retrosplenial cortices), although it was detected in a few PA-, CB-, VIP-, CCK-, SS-, or NPY-immunoreactive nonpyramidal neurons in the piriform, entorhinal, and hippocampal cortices. PA- and CB-positive neurons have been reported to constitute the major population of GABAergic neurons in the cerebral cortex. Thus, the present results, together with the previous reports, suggest that most GABAergic, cholinergic and peptidergic nonpyramidal neurons in the neo- and mesocortex do not contain glutaminase.

Animals↗

PubMed1992

Distribution of the mRNA for a metabotropic glutamate receptor (mGluR1) in the central nervous system: an in situ hybridization study in adult and developing rat.

Distribution of the mRNA for a metabotropic glutamate receptor (mGluR1), which is linked to phosphoinositide (PI) hydrolysis, was investigated in adult and developing rat central nervous system (CNS) by in situ hybridization. Transcripts of mGluR1 were specifically localized to neurons and widely distributed throughout the adult rat brain. Most intensely labeled neurons were Purkinje cells of the cerebellum, mitral and tufted cells of the olfactory bulb, and neurons in the hippocampus, lateral septum, thalamus, globus pallidus, entopeduncular nucleus, ventral pallidum, magnocellular preoptic nucleus, substantia nigra, and dorsal cochlear nucleus. Moderately labeled neurons were seen in high density in the dentate gyrus, striatum, islands of Calleja, superficial layers of the retrosplenial, cingulate and entorhinal cortices, mammillary nuclei, red nucleus, and superior colliculus. In the developing rat brain, the level of mGluR1 expression gradually increased during early postnatal days in accordance with the maturation of neuronal elements. These results show prominent expression of mGluR1 in the major targets of putative glutamatergic pathways and unique distribution pattern of mGluR1 distinct from those reported for ionotropic subtypes of glutamate receptors, suggesting specific roles of mGluR1 in the glutamatergic system.

Aging↗

PubMed1992

Non-dopaminergic neurons in the substantia nigra project to the reticular formation around the trigeminal motor nucleus in the rat.

The dorsolateral part of the substantia nigra (SN) of the rat was observed to send projection fibers to the reticular formation (RF) around the trigeminal motor nucleus (Vm), bilaterally with a clear-cut ipsilateral dominance, by the anterograde and retrograde tracing techniques with PHA-L and WGA-HRP. A combination of retrograde tracing and immunohistochemistry for tyrosine hydroxylase (TH) revealed that no SN neurons sending their axons to the RF around the Vm showed TH-like immunoreactivity.

Animals↗

PubMed1992

Molecular characterization of a novel metabotropic glutamate receptor mGluR5 coupled to inositol phosphate/Ca2+ signal transduction.

A cDNA clone for a new metabotropic glutamate receptor, mGluR5, was isolated through polymerase chain reaction-mediated DNA amplification by using primer sequences conserved among the metabotropic glutamate receptor (mGluR) family and by the subsequent screening of a rat brain cDNA library. The cloned receptor consists of 1171 amino acid residues and exhibits a structural architecture common to the mGluR family, possessing a large extracellular domain preceding the seven putative membrane-spanning segments. mGluR5 shows the highest sequence similarity to mGluR1 among the mGluR members and is coupled to the stimulation of phosphatidylinositol hydrolysis/Ca2+ signal transduction in Chinese hamster ovary cells transfected with the cloned cDNA. This receptor also resembles mGluR1 in its agonist selectivity and antagonist responses; the potency rank order of agonists for mGluR5 was determined to be quisqualate greater than L-glutamate greater than or equal to ibotenate greater than trans-1-aminocyclopentane-1,3-dicarboxylate. Blot and in situ hybridization analyses indicated that mGluR5 mRNA is widely distributed in neuronal cells of the central nervous system and is expressed differently from mGluR1 mRNA in many brain regions. This investigation thus demonstrates that there is an additional mGluR subtype which closely resembles mGluR1 in its signal transduction and pharmacological properties and is expressed in specialized neuronal cells in the central nervous system.

Amino Acid Sequence↗

PubMed1992

An electron microscopic study of neurosecretion in the cerebral ganglion of the earthworm.

Non-synaptic, exocytotic release of neurosecretory granules in cerebral ganglion neurons was observed electron microscopically in 3 species of the oligochaete annelids Aporrectodea caliginosa, Octolasion cyaneum and Lumbricus terrestris. In addition to the features indicating exocytotic release of neurosecretory granules into perineuronal space, possible features of neurosecretion into blood vessels were seen within the cerebral ganglion. Axon terminals in synaptic contact with perikaryal profiles of cerebral ganglion neurons were also found.

Animals↗

PubMed1992

Topographic projections from the subiculum to the limbic regions of the medial frontal cortex in the cat.

When WGA-HRP was injected into the subicular cortex of the ventral hippocampal formation (Hv) in the cat, terminal labeling was seen ipsilaterally in the caudoventral parts of the medial frontal cortex: in the infralimbic cortex (area 25), ventral part of the prelimbic cortex (area 32), and caudoventral part of the orbitofrontal cortex (area 12). The terminal labeling was observed in all cortical layers except layer 1. The cells of origin of these projections were then confirmed to be pyramidal neurons in the subiculum of the Hv. The projections were topographically organized: the temporal-septal shift along the long axis of the subiculum of the Hv corresponded to a rostroventral-caudodorsal shift in the anterogradely labeled limbic regions of the medial frontal cortex.

Animals↗

PubMed1992

The phosphorylated and/or sulfated structure of the carbohydrate-protein-linkage region isolated from chondroitin sulfate in the hybrid proteoglycans of Engelbreth-Holm-Swarm mouse tumor.

The structure of the linkage region of chondroitin sulfate chains attached to the hybrid proteoglycans of the Engelbreth-Holm-Swarm mouse tumor was investigated. The peptidoglycan fraction which contains oversulfated chondroitin sulfate rich in the GlcA beta 1-3GalNAc-4,6-diO-sulfate unit and undersulfated heparan sulfate rich in GlcA beta 1-4GlcNAc and GlcA beta 1-4GlcN-2N-sulfate units was isolated after exhaustive protease digestion of the acetone powder of the tumor tissue, (GlcA, glucuronic acid; GalNAc, 2-deoxy-2-N-acetylamino-D-galactose). Glycosaminoglycans were released by beta-elimination using NaB3H4 and digested with chondroitinase ABC. The linkage region fraction was separated from heparan sulfate by gel filtration and fractionated by HPLC on an amine-bound silica column. Six radiolabeled compounds (L1-L6) were obtained and structurally analyzed by cochromatography with authentic hexasaccharide alditols recently isolated by us from the linkage region, and by digestion using chondroitinase ACII, alkaline phosphatase and beta-galactosidase in conjugation with HPLC. These compounds shared the conventional hexasaccharide backbone structure: delta GlcA beta 1-3GalNAc beta 1-4GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl-ol, (delta GlcA, delta 4.5-GlcA or D-gluco-4-enepyranosyluronic acid). L1 was not sulfated or phosphorylated. L2 and L4 were monosulfated at C-6 and C-4 of the GalNAc residue, respectively. Upon alkaline phosphatase digestion, L3, L5 and L6 were converted to L1, L2 and L4, respectively. Analysis of the periodate oxidation products indicated that the phosphate group in L3, L5 and L6 is located at C-2 of Xyl-ol. These results suggest that Xyl-2-O-phosphate is associated with both 4-O-sulfated and 6-O-sulfated GalNAc units and does not directly determine the sulfation pattern of chondroitin sulfate.

Animals↗

PubMed1992

Direct projections from the globus pallidus to the midbrain and pons in the cat.

Employing the anterograde and retrograde axonal tracing techniques with Phaseolus vulgaris leucoagglutinin and cholera toxin B subunit, we demonstrated direct projections from the globus pallidus (GP) to the midbrain and pons in the cat. Cells of origin of these projections were localized in the caudal 2/3 of the GP, and their major target sites included the peripeduncular region, nucleus of the brachium of the inferior colliculus, para-lateral lemniscal zone, nucleus sagulum, external and pericentral nuclei of the inferior colliculus, and cuneiform nucleus. A combination of retrograde axonal tracing and immunohistochemistry for choline acetyltransferase revealed that GP neurons giving rise to such descending projections were primarily non-cholinergic.

Animals↗