| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1986: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
The evidences were obtained that protein kinase C (C-kinase) was involved in the exocytosis of neurotransmitter release from the central and peripheral nervous tissues. Monoclonal antibodies against C-kinase were obtained by immunizing C-kinase which was purified from the soluble fraction of the brain. Using these monoclonal antibodies, C-kinase-positive immunoreactin was present in the hippocampus, olfactory bulb and cerebellar cortex. The autoradiograms of [^3H]4<beta>-phorbol 12,13-dibutyrate ([^3H]PDBu), a ligand of potent phorbol ester, in brain was uneven with the highest levels in etlencephalon, which resembles C-kinase immunoreactivities. TPA, a phorbol ester which activates C-kinase potentiated the Ca^<2+>-dependent release of acetlycholine (ACh), noradrenaline (NA) and <gamma>-aminobutyric acid (GABA) from central and peripheral nervous systems. The potentiation of NA release by the activation of C-kinase was found to be inhibited by by the forskolin-induced activation of cyc
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lic AMP-A-kinase system. This indicates that the Ca^<2+>-dependent release of neurotransmitters is regulated by the diacylglycerol-C-kinase system and the cyclic AMP-A-kinase system. Recent analysis of the complementary DNA clones has revealed that at least three different genes appear to be expresses in the brain and at least four subspecies (alpha), <beta>I, <beta>II and <gamma> of the enzyme have been identified. In order to investigate which subspecies of C-kinase are involved in the release of neurotransmitters, we examined the localization of subspecies of C-kinase in the brain. <gamma>-Type of C-kinase was found to be present in the nerve terminals of purkinje cells which contain GABA and project to the deep cerebellar nucleus. The activation of C-kinase by TPA potentiacted the stimulation-induced release of GABA from the deep cerebellar nucleus. Thus, the -type of C-kinase may be involved in the release of ygaba from the nerve terminals. The physiological role of C-kinase may be elucidated by the electron microscopic examination of the localization of each subtype of C-kinase. Less
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