| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Sei Kyushu University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (80225515)
HIRANO Katsuya Kyushu University, Faculty of Medicine, Lecturer, 医学部, 講師 (80291516)
NISHIMURA Junnji Kyushu University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (90237727)
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| Research Abstract |
For the prevention of coronary vasoconstriction and intimal thickening, following studies were performed : In smooth muscle and endothelial cells, (1)the characterization of the intracellular signal transduction networks, (2) the elucidation of the relationships between cell cycle progression and the cellular functions, and (3) the elucidation of the relationships between signaling factors and cellular functions/structures. (4) The clinical applicatopn of the results of these experimental studies.
Following results/conclusions were obtained : (1) We have developed a method to monitor [Ca]i changes of the vascular strips, which was reported as the chapter "Measurement of [Ca]i in smooth muscle strips using front-suface fluorimetry" (P269-P277) by H.Kanaide in the book "Calcium signaling protocols" edited by David G.Lambert, which is the Vol.114, of series of "Methods in Molecular Biology" (Humana Press Inc., Totowa, NJ, USA, 1999). Using this technique and skinning of cell membrane, we have characterized the signal transduction system of smooth muscle and endothelial cells. In particular, we have characterized the Ca-sensitivity of the contractile apparatus in smooth muscle cells and relationships between [Ca]i and NO production in endothelial cells. (2) We found that types of Ca-channels(T,L) varied during the progression of cell cycles in smooth muscle cells. There appeared submembranous localization of myosin phosphatase at the phase of contact inhibition of the growth in endothelial cells. (3) Using DNA mutants, we have characterized the relationships between protein structure and funtion of calmodulin and 130 and 20 KDa subunits of myosin phosphatase. (4) There is a mutation of Trp64Arg in DNA of beta 3 adrenoceptors among the patients with ischemic heart disease. However, the clinical application of the results of the present study has remained to be the next problem which confronts us.
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