The role of Ca^<2+>-dependent protein kinases in central nervous system in health and diseases.

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To elucidate the functions of Ca^<2+>-dependent protein kinases in central nervous system in health and diseases, biochemical and pharmacological approaches were performed in this study. First, we have developed the subtype specific enzyme immunoassay (EIA) using three anti-protein kinase C (PKC) isozymes monoclonal antibodies. The EIA showed the quantitative analysis and subtype-specific expression of PKC in CNS tumor cells including glioblastoma and neuroblastoma cells. The results suggested that PKC isozymes have a cell-type specificity in CNS tumors, and that detection of PKC isozymes will aid in understanding the diverse effects of PKC and also to diagnose tumors derived. from glia or neuronalcells. In addition, we produced antibodies against the novel PKCS, and examined tissue and cell-type specific expression of the novel PKCS.Second, we have newly synthesized specific inhibitors of various protein ldnases including Ca^<2+>/calmodulin protein kinase II (CaM Idnase 11), cAMp-dependent protein kinase (A-kinase) and casein kidase I. KN-62 is specific CaM kinase 11 inhibitor and dose not inhibit other kinase activities. Furtennore, KN-62 inhibit the releasing of GABA and long term potentiation in central nervous system suggesting that the compound is a useful tool for elucidation of the functions of CaM kinase H in many tissues. H-89 (A-kinase inhibitor) also inhibited neurite growth n PC- 12 cells induced by folskolin, and induction of immediately early gene.We could determined the cell-type specific expression of PKC isozymes and developed specific inhibitors of various protein kinases. We should try on next steps to elucidate the functions of these protein Idnases using specific inhibitors.