Changes in Ca^<2+> Signaling and Contractile Protein Isoforms in Smooth Muscle Cells from Guinea Pig Ileum during Culture
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概要
- 論文の詳細を見る
Single smooth muscle cells(SMCs)isolated from guinea pig ileum using collagenase and papain were cultured on laminin-coated dishes in MEM containing fetal calf serum. Temporal changes in intracellular calcium ion concentration in response to carbachol and to ATP were investigated using fluo-3/AM and fluorescence microscopy. It was observed that carbachol caused an increased intracellular calcium ion in freshly isolated single SMCs but a reduced or negative response of cultured SMCs before confluence. On the other hand, ATP was observed to cause an increase in the calcium ion content of SMCs throughout the culture. SDS-PAGE and Western blot analyses revealed changes in the expression of contractile proteins as follows. l-Caldesmon and non-muscle type myosin heavy chain(NMHC)(considered to be marker molecules for dedifferentiation in smooth muscle cells)and non-muscle type tropomyosin were not observed in freshly isolated single SMCs. l-Caldesmon and NMHC appeared in the cultured SMCs within 2 days and the tropomyosin isoform was observed 6 days following seeding. Simultaneously, smooth muscle type myosin heavy chain(SMHC)decreased strikingly and the 41 kDa tropomyosin monomer was lost. The content of α-action decreased gradually to a minimum on day 6 when non-muscle type tropomysin appeared, and the cells began to proliferate rapidly. These results suggest that the loss of contractility in cultured smooth muscle cells is more closely related to changes in contractile protein profiles than to receptor-mediated signal transduction and that in addition to NMHC and l-caldesmon, non-muscle type tropomyosin may be useful as a marker molecule for de-differentiation of smooth mescle cells.
- 日本平滑筋学会の論文
著者
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OHATA Hisayuki
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
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MOMOSE Kazutaka
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
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IIJIMA Michiko
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
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YAMAMOTO Junko
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
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Ohata H
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
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Ohata Hisayuki
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
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Iijima Michiko
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
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Momose K
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
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Momose Kazutaka
Department Of Phamacology School Of Pharmaceutical Sciences Showa University
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TAKADA Noriko
Department of Pharmacology, Schoolof Pharmaceutical Sciences, Showa University
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Takada Noriko
Department Of Pharmacology Schoolof Pharmaceutical Sciences Showa University
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Yamamoto Junko
Department Of Obstetrics And Gynecology Okayama University Medical School
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Yamamoto Junko
Department Of Pharmacology Schoolof Pharmaceutical Sciences Showa University
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Yamamoto Junko
Department Of Chemistry Faculty Of Science Shizuoka University
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