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H Higashida

Publications and source records attributed to H Higashida.

139 records · Page 8Linked to original sources
PubMed1981

Regulation of neuron-specific enolase in NG108-15 hybrid cells and C6BU-1 glioma cells.

Distribution of three isoenzymes of brain enolase (2-phospho-D-glycerate hydro-lyase, EC 4.2.1.11) (alpha alpha, alpha gamma and gamma gamma forms) in clonal cell lines of neuroblastoma (NS20Y and N18TG-2), glioma (C6BU-1), and hybrid cells NG108-15, NCB20, Nbr10A, Nbr20A, N4G-B-a and N4G-C-a) was examined with a sensitive enzyme immunoassay system, that uses a rabbit antibody to rat brain enolase alpha alpha or gamma gamma. All cell lines tested were found to possess the enolase which contains gamma subunit (a neuron-specific protein), although the alpha alpha enolase (non-neuronal enolase) was the dominant from in these cells. A clonal rat glioma (C6BU-1) cell contained about 40, 1 and 0.07 microgram/mg protein of alpha alpha, alpha gamma and gamma gamma enolases, respectively, at the confluent stage. Inclusion of 1 mM dibutyryl cyclic AMP or 10 micrometers prostaglandin E1 plus 1 mM theophylline in the culture medium of a hybrid cell (NG108-15, mouse neuroblastoma x rat glioma) resulted in a more than 2-fold increase in the concentrations of alpha gamma and gamma gamma in the cell within a few days, with little change in the alpha alpha enolase concentration. A similar increase in the concentration of gamma subunit by the nucleotide (but not by prostaglandin E1 plus theophylline) was also observed in the glioma cell (C6BU-1) line. The results suggest that the gamma subunit or the neuron-specific protein can be regulated in NG108-15 and C6BU-1 cells in a cyclic AMP-dependent fashion.

Animals↗
PubMed1981

Proliferation and synapse formation of neuroblastoma glioma hybrid cells: effects of glia maturation factor.

Glia maturation factor (GMF), extracted from bovine brain, stimulated DNA synthesis and proliferation of glioma cells and hybrid cells derived from glioma and neuroblastoma cells (NG108-15), but had no effect on neuroblastoma cells. The synapse formation of NG108-15 cells with rat striated myotubes was lower in the presence of GMF than the control and also lower after treatment with prostaglandin E1 (PGE1) plus theophylline, indicating that GMF did not induce functional differentiation of NG108-15 cells. The results show that expression of mitogenic action for GMF in the hybrid cells is a property derived from the glioma parent, and that NG108-15 is therefore an excellent model for studying glial-neuronal interactions.

Animals↗
PubMed1981

Characterization of chick gizzard extract that promotes neurite outgrowth in cultured ciliary neurons.

Chicken gizzard extract contains a macromolecule(s) that promotes the neurite outgrowth of dissociated neurons from the ciliary ganglia (CG) of chick embryos. The factor in gizzard extract was partially purified and estimated to be about 12S (M.W. 200,000-300,000) on sucrose density gradient centrifugation. The neurite outgrowth of CG neurons by the factor strictly depends on the embryonal age. The maximal neurite outgrowth was observed when CG neurons were dissociated from the embryos younger than 10 days. After that time the response of CG neurons to the factor rapidly declined and was almost lost at day 14. The amount of factor in the gizzard began to increase rapidly from 12-day-old embryo and reached the maximal level at day 16, and thereafter a fairly steady level was maintained. When CG neurons were cocultured with rat myotubes, the ratio of muscle cells with synaptic responses (miniature end-plate potentials) was significantly higher in the presence of gizzard factor than its absence. The results suggest that this factor acts as an external signal on CG neurons to form synaptic connections in vivo.

Animals↗
PubMed1981

[Neuropharmacology of cultured cells: studies on receptors, synapses and neuronal function of clonal cells (author's transl)].

This review concerns neuropharmacological properties of clonal cells from tumors in the nervous system and muscle, and/or somatic hybrid cells derived from these clonal preparations. These cells grow well under conditions of culture and show neuronal characteristics identical to those seen in normal cells. Observations of these clonal cells contribute to studies on development, differentiation, synaptogenesis and cellular recognition in the nervous system. Analysis of eukaryotic genes also enables investigations on genetic control mechanisms by which highly differentiated neuronal functions are expressed.

Adenylyl Cyclases↗
PubMed1979

Adenylate cyclase and acetylcholine release regulated by separate serotonin receptors of somatic cell hybrids.

Serotonin activates adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] of NCB-20 neuroblastoma--brain hybrid cells with an activation constant of 530 nM, but has little or no effect on cellular cyclic AMP or cyclic GMP content of NIE-115 neuroblastoma or NG108-15 hybrid cells. In homogenates of NCB-20 hybrid cells, lysergic acid diethylamide stimulates adenylate cyclase activity (Kact = 12 nM) and partially inhibits (Ki = 10 nM) the stimulation of adenylate cyclase activity by serotonin. No desensitization was detected of serotonin receptors coupled to adenylate cyclase. Serotonin also depolarizes NCB-20, NG108-15, and NIE-115 cells and increases acetylcholine release. Serotonin receptors mediating depolarizing responses desensitize rapidly and reversibly, and the depolarizing effects of serotonin are neither mimicked nor inhibited by lysergic acid diethylamide. These results indicate that (i) NCB-20 cells possess at least two species of serotonin receptors, which independently regulate cellular functions, (ii) activation of adenylate cyclase does not directly affect membrane potential or acetylcholine release, and (iii) serotonin-dependent cell depolarization does not affect cyclic AMP or cyclic GMP synthesis in the cell lines tested.

Acetylcholine↗
PubMed1977

Spreading depression in isolated carp retina.

(1) Spreading depression (SD) could be elicited in isolated carp retina by KCI application, the concomitants of which were similar to those described in other vertebrates. (2) The threshold for generating SD was greatly reduced by brief immersion of the retina in low ci-ringer's solutions. The properties of SD waves were almost the same with treated and untreated retinas, except for intervals. (3) Extracellular negative potential shifts during SD, averaging 4.6 mV in amplitude and 27 sec in duration, were recorded in whole retinal layers with the maximum amplitude about at the inner plexiform layer. (4) The PIII potential of the local electroretinogram was virtually unaffected by SD. (5) Both L-and C-type S-potentials could be evoked with increase of 20-40% in amplitude around the peak of slow membrane depolarizations (mean value of maximal amplitude 5.8 mV) during SD in horizontal cells. (6) Increase in spike number was observed in both on-and off-center ganglion cells before and after the spike cessation during SD in the untreated retina. However, the off-discharges, which were a unique response to light in the immersed retina, only decreased during SD.

Animals↗
PubMed1975

Mitochondrial malate dehydrogenase of bovine cerebrum. Characterization and mechanisms of inhibition by silver ions.

Attempts were made to characterize mitochondrial malate dehydrogenase [L-malate: NAD+ oxidoreductase, EC 1.1.1.37] (M-MDH) purified from bovine cerebrum and to elucidate the mechanisms responsible for inhibition of the enzymic activity by Ag+. The molecular weights of the native enzyme and its subunits were 54,000-55,000 and 30,000-32,000, respectively. In general, the physiochemical and catalytic properties of bovine cerebral M-MDH was not very different from those of other corresponding mammalian enzymes. Incubation of the enzyme with Ag+ caused the loss of equivalent amounts of sulfhydryls with a parallel decrease of the enzymic activity. When the enzyme was exposed to 2-, 3.5-, and 5-fold molar excesses of Ag+, the enzymic activity showed an initial rapid fall and a subsequent slow restoration to a partially inactivated level (60-70, 45-50, and 15-20% of an untreated control, respectively), while the alpha-helical content of the enzyme fell exponentially with time. A 7-fold molar excess of Ag+ reduced both the enzymic activity and the alpha-helical content to a much greater degree and no restoration of the enzymic activity was observed. The Km values of Ag+-inactivated enzyme for NADH and oxaloacetate were the same as those of the native enzyme. The data suggest that Ag+ could inhibit enzymic activity both by reducing the structural regularity of the enzyme molecule and by attacking sulfhydryl groups necessary for the catalytic activity of bovine cerebral M-MDH.

Amino Acids↗
PubMedDate not supplied

Cyclic ADP-ribose as a second messenger revisited from a new aspect of signal transduction from receptors to ADP-ribosyl cyclase.

Cyclic ADP-ribose (cADPR), an endogenous modulator of ryanodine receptor Ca(2+)-releasing channels, is found in various tissues. Cytosolic injection of cADPR induces an elevation of intracellular Ca(2+) concentrations or potentiates Ca(2+) increases. cADPR facilitates neurotransmitter or insulin release and modifies ionic currents. cADPR is synthesized by ADP-ribosyl cyclase and is metabolized by cADPR hydrolase. ADP-ribosyl cyclase activity is up-regulated by nitric oxide/cyclic GMP-dependent phosphorylation or receptor stimulation via G-proteins within membranes. These findings suggest that cADPR is a second messenger in cellular Ca(2+) signaling. However, many intriguing issues remain to be addressed before this identity is confirmed.

ADP-ribosyl Cyclase↗
PubMedDate not supplied

Two polyphosphatidylinositide metabolites control two K+ currents in a neuronal cell.

Hydrolysis of the membrane phospholipid phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) produces two prospective intracellular messengers: inositol 1,4,5-trisphosphate (InsP3), which releases Ca2+ from intracellular stores; and diacylglycerol (DG), which activates protein kinase C. Here we show how the formation of these two substances triggered by one external messenger, bradykinin, leads to the appearance of two different sequential membrane conductance changes in the neurone-like NG108-15 neuroblastoma-glioma hybrid cell line. In these cells bradykinin rapidly hydrolyses PtdIns(4,5)P2 to InsP3 and DG, raises intracellular Ca2+ and hyperpolarizes then depolarizes the cell membrane. By voltage-clamp recording we show that the hyperpolarization results from the activation pharmacologically-identifiable species of Ca2+-dependent K+ current. This is also activated by intracellular injections of Ca2+ or InsP3 so may be attributed to the formation and action of InsP3. The subsequent depolarization results primarily from the inhibition of a different, voltage-dependent K+ current, the M-current that is also inhibited by DG activators. Hence we describe for the first time a dual, time-dependent role for these two intracellular messengers in the control of neuronal signalling by a peptide.

Animals↗