| Project/Area Number |
15510185
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Kwansei Gakuin University |
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Principal Investigator |
SUZUKI Shintaro Kwansei Gakuin University, School of Science and Technology, Professor, 理工学部, 教授 (20128432)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | cardiomyocyte / cell adhesion / intercalated disk / adherens junction / desmosome / cadherin / integrin / actin / アドヒレンスジャンクション |
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| Research Abstract |
Formation of intercalated discs of cardiac muscle was investigated by using a cell culture system. First, a cell culture system of cardiomyocytes was developed. Rat hearts from 19-day embryos were dissected and the cardiomyocytes were dissociated with collagenase. Then, the cardiomyocytes were purified by Percoll density-gradient centrifugation. The resultant cells were cultured on cover glasses coated with collagen in DMEM medium containing horse serum and b-FGF. After 10-14 days, cardiomyocytes became to align and intercalated disc-like cell adhesion structure was gradually formed at the ends of the elongated cells. Inhibition of fibroblast growth with AraC helped long time culture of cardiomyocytes and facilitated the formation of intercalated discs-like structure. Rat serum was as effective as horse serum, but bovine serum was not for the formation of intercalated disc-like structure. TGF-β enhanced the beating of cultured cardiomyocytes, b-FGF facilitated the alignment of the cells and IGF stimulated the formation of actin fibers. The formation of intercalated disc-like structure was not dependent of cell-shape or cell alignment, but required differentiation of cardiomyocytes. Furthermore, the intercalated discs-like structure was formed at the attachment sites of actin fibers to plasma membrane, suggesting a close relationship between them. Microtubules seemed to stimulate the alignment of actin fibers and the localization of intercalated disc-like structure. The next important question is the regulation of intercalated disc formation.
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