1992 Fiscal Year Final Research Report Summary
Analysis of molecular mechanism in Ca^<2+>-independent vasocontraction
| Project/Area Number |
03454252
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Mie University |
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Principal Investigator |
NAKANO Takeshi Mie Univ., Faculty of Medicine, Professor, 医学部, 教授 (60111879)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Masaaki Univ., Faculty of Medicine, Assistant Professor, 医学部, 助手 (00223181)
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| Project Period (FY) |
1991 – 1992
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| Keywords | vascular smooth muscle / calcium / myosin light chain / phosphatase / phorbol ester / endothelin |
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| Research Abstract |
In vascular smooth muscle, some agonists induce sustained contraction even in the absence of extracellular Ca^<2+> without increases in intracellular free Ca^<2+> level ([Ca^<2+>]i). To assess the Ca2+-independent contractile mechanism(s), we examined the effects of various agents on intact vascular smooth muscle (rat thoracic aorta strips) in Ca^<2+>-free buffer, focusing on [Ca^<2+>]i, myosin light chain (MLC) and MLC-phosphatase activity. Endothelin-1 (ET-1), phorbol 12-myristate 13-acetate (PMA), phorbol 12,13-dibutyrate (PDB) and caliculin-A produced sustained contractions of Ca^<2+>-depleted rat thoracic aorta strips, prepared by repeated applications of norepinephrine or caffeine in Ca^<2+>-free buffer. After 10 min incubation in Ca^<2+>-free buffer, norepinephrine induced a typical phasic contraction and a transient increase in [Ca^<2+>]i, which measured with fura 2. On the other hand, PMA and PDB induced sustained contraction in Ca^<2+>-free buffer in [Ca^<2+>]i. The PDB-induced contraction was associated with the phosphorylation of 20-kDa MLC. Two-dimentional phosphopeptide mapping of 20-kDa MLC revealed that about nine-tenths of the phophopeptides was derived from MLC kinase-catalyzed reaction and about one-tenth was due to phosphorylation by protein kinase C. MLC-phosphatase was purified from chicken gizzard. The enzyme consisted of 58-kDa regulatory subunits and 38- kDa catalytic subunit. Although ET-1 and PDB did not show any significant effects on the purified phosphatase activity, MLC-phosphatase activity of intact aorta strip was inhibited by the treatment with PDB. These results suggest that the regulation of MLC-phosphatase in vascular smooth muscle may play important roles in the Ca^<2+>-independent contraction.
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Research Products
(16 results)
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[Publications] H. Itoh, G.R. Bolt, K.L.: "MacCannel and T. Nakano, Interference of a selective mesenteric vasodilator peptide, urotenin I, with adrenergic pressor responses." alpha-adrenoceptors (edited by M. Fujiwara, T. Sugimoto, K. Kyuya, Excerpta Medica, Tokyo, Japan). 230-233 (1992)
Description
「研究成果報告書概要(欧文)」より
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[Publications] S. Okubo, F. Erdori, M. Ito, K. Ichikawa, T. Konishi, T. Nakano, T. Kawamura: "D.L. Brautigan and D.J. Hartshorne, Characterization of a myosin-bound phosphatase from smooth muscle." Adv. Prot. Phosphatase 7 (edited by W. Merlevede, Leuven Univ. Press, Leuven, Belgium).
Description
「研究成果報告書概要(欧文)」より
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