| Project/Area Number |
61571066
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Showa University |
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Principal Investigator |
MOMOSE KAZUTAKA School of Pharmaceutical Sciences, Showa University, Professor, 薬学部, 教授 (80004597)
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| Co-Investigator(Kenkyū-buntansha) |
TOMOKO KAMISHIMA School of Pharmaceutical Sciences, Showa University, Research Asistant, 薬学部, 助手 (70177575)
CHIEKO YOSHIDA School of Pharmaceutical Sciences, Showa University, Research Assistant, 薬学部, 助手 (50166956)
IKUO MARUYAMA School of Pharmaceutical Scineces, Showa University, Research Assistant, 薬学部, 助手 (20053994)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | guinea pig / taenia coli / single cells / smooth muscle / Ca-contraction / cch収縮 |
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| Research Abstract |
We established isolation procedures of single smooth muscle cells and investigated the physiological, pharmacological and morphological properties of the isolated cells. Results are as follows:
1. Isolation procedures of single smooth muscle cells. For isolation of smooth muscle cells, commercially available collagenase preparations have been used, but, the specific activities were dependent on the preparations. Therefore we tried combination of highly purified collagenase and papain and it resulted in high yield and good viability of the cells. Cells isolated under the conditions possessed the contractility.
2. Morphology of single smooth muscle cels. Single cells were examined with use of electron microscopes in order to check the damages of membranes during the isolation [rocedures. The sizes of isolated cells were smaller than those in the tissue, and the changes in morphological aspects of organelles and contractile systems in the cells during contraction were quite different from those in the tissues. But, significant damages were not found in the membranes.
3. Physiological and pharmacological properties and application of single cells to smooth muscle researches. The isolated cells were well responsive to agonists and this responses were inhibited by the specific antagonists. So, the pharmacological and physiological properties must be well preserved after the isolation. We also tried to apply the single cells to investigate calcium movement in the cytoplasm of the smooth muscle cells during the contraction, with use of jura 2, a calcium sensitive fluorescence, in the presence of DFP, a powerful cholinesterase inhibitor. By this procedure, changes in calcium concentrations in the cells were easily determined.
On finishing the research program, we concluded that the single smooth muscle cells must be able to be applied for smooth muscle research as a simple model of smooth muscle.
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