| Project/Area Number |
06044037
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Chiba University |
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Principal Investigator |
SHIMADA Yutaka Chiba University, Professor, 医学部, 教授 (70009116)
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| Co-Investigator(Kenkyū-buntansha) |
PEXIEDER Tomas University of Lausanne, 準教授
PERRIARD Jea スイス工科大学, 教授
KOMIYAMA Masatoshi Chiba University, 医学部, 助手 (70175339)
TOYOTA Naoji Chiba University, 医学部, 講師 (00188822)
PEXIEDER Jean-Claude Swiss Federal Institute of Technology
PEXIEDER To ローザンヌ大学, 医学部, 準教授
PERRIARD Je スイス工科大学, 教授
立木 幸敏 国際武道大学, 体育学部, 助手 (20255178)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 1996: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1995: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1994: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | isoform / myosin alkali light chain / troponin I / myofibril / heart / retinoic acid / morphogenesis / 分子種 / エピトープタギング / 心筋細胞 / 筋原線維形成 / アクチン / ミオシン / コネクチン / ネブリン / 細胞内導入 |
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| Research Abstract |
1. Mechanisms of myofibrillar protein isoform changes during development
Various combinations of two different myosin alkali light chain (LC) isoform cDNAs tagged with either VSV or mT antigen were co-exapressed in cultured cardiomyocytes from rat and chicken ventricles. Expressed isoproteins were detected using antibodies against the tags and their sorting patterns were analyzed by conforcal microscopy. The sorting specificity of LC isoforms to sarcomeres was shown to increase in the order of following developmental expression sequence : nonmuscle, slow skeletal muscle, ventricular, and fast skeletal muscle type. This suggests that during myofibrillogenesis isoforms with progressively higher binding affinities to myofibrils efficiently replace previous proteins, ensuring rapid exchange of isoforms without an extraordinary increase in protein synthesis. Expression of chimeric cDNAs revealed that the second EF-hand in the N-terminal lobe of each isoprotein is responsible for the isoform-
… More
specific sorting pattern. Although this part is not included in the key contact (s) with the myosin heavy chain, amino acid differences in this domain might affect the relative disposition of neighboring domains, thereby influencing the precise association of the key contact (s).
2. Behavior of cardiac and skeletal muscle type isoproteins during myofibrillogenesis
Cardiac and skeletal (breast) muscle troponin Is (CTnI and FTnI,respectively) and their mutants (chimeric and truncated TnIs) were force expressed in cultured muscle cells to examine incorporation mechanism of TnI isoforms into myofibrils (Mfs). Since CTnI was not incorporated into breast Mfs, sorting mechanism of CTnI appeared to exist in breast MFs. Analysis of chimeric and truncated TnIs showed that TnIs were composed of two functionally distinct domains : C-terminal domain which binds to Mfs and N-terminal domain regulating the strength of binding affinity and specificity to cardiac and breast MFs.
3. Morphogenesis and teratogenesis of the heart
The cardiac wall in embryos was investigated with the scanning electron microscope. The different surface characteristics of the four heterotypic cells that constitute the embryonic heart were demonstrated. The epicardial cells migrate from the mesothelium of the sinus venousus, encircled the ventricular wall, then extended cranically/caudally to unsheathe the entire heart usrface. Incidence of cardiac malformation was very high if retionic acid was administered into pregnant mice on 8ed day at the dose of 20 mg/kg body weight. Abnormalities observed are : TGA,DORV,VSD,A-loop and L-loop. Less
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