| Project/Area Number |
60480098
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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 |
General anatomy (including Histology/Embryology)
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| Research Institution | Chiba University |
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Principal Investigator |
SHIMADA Yutaka Chiba University School of Medicine,Professor, 医学部, 教授 (70009116)
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| Co-Investigator(Kenkyū-buntansha) |
石出 猛史 千葉大学, 医学部, 助手 (60184545)
TOYOTA Naoji Chiba University Schoolof Medicine,ResearchAssociate, 医学部, 講師 (00188822)
ISOBE Yuji Chiba University School of Medicine,Research Associate, 医学部, 助手 (10159816)
門田 朋子 千葉大学, 医学部, 助教授 (00089864)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1985: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | Myofibril / Cytoskeleton / Cell cycle / Immunofiuorescence / Freeze-Fracture deep-etch Scanning electron microscopy / Focal contact / 遺伝子 / 筋細胞 / ミトコンドリア / 小胞体 / T管 / アクチンフィラメント / 中間径フィラメント / 細胞分裂周期 |
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| Research Abstract |
Our project was to study the differentiation of cytoskeletons in embryonic muscle cells by using morphological,biochical,immunological and gene-technological methods.
(1)Differentiation of cytoskeletons:The freeze-fracture, deep-etch replica method revealed that embryonic skeletal muscle cells in vitro contained two cytoskeletal domains: subsarcolemmal and endoplasmic cytoskeletal networks.The former consisted of mainly actin filaments;the latter intermediate filaments. 2-4 nm filaments interconnected those cytoskeletons with the inner surface of the cell memberane and with myofibrils. These cytoskeletal structures appear to play some roles in maintaing the cellular shape as well as positioning myofibrils within the cells.
(2) Cytoskeleton and myofibrillogenesis:Embryonic cardiac myocytes in vitro were stained with phalloidin and antibodies against troponin C, myoshin heavy chain, alpha-actinin, vinculin and fibronectin, and examined with the fluorescence and interference reflection micr
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oscopes. Thrminals of early myofibrils were associated with alpha-actinin, vinculin, fibronectin, and cell-substrate focal contacts. It appears that the formation of focal contacts and the accumulation of the adhesion plaque proteins at these sites must involve a polarization of molecules within the cell; myofibrillogenesis occurs along the inferred polarization lines generated between the focal cell contact sites.
(3) The cDNA of troponin C and I and gene expresion:We have isolated cDNA clones coding for chicken cardiac troponin C and I(CTnC and CTnI,respectively).Nucleotide sequence analysis revealed that CTnC had a full length cDNA;CTnI contained approximately 78% of amino acid sequence of TnI.Total RNAs isolated_<32>from caridec and skeletal muscles at different developmental stages were hybridized to P-labeled CTnC and CTnI.Sifnals of TnC mRNA were seen in RNA from embryonic and adult cardiac muscles as well as embryonic skeletal muscle.CTnI hybridized with RNA from embryonic and adult cardiac muscle. These results are consistent with the data by immunohistochemistry. Less
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