ANALYSIS OF GENE FUNCTIONS USING AMPHIBIAN
| Project/Area Number |
12680701
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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 |
Cell biology
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| Research Institution | SAITAMA MEDICAL SCHOOL |
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Principal Investigator |
KOMAZAKI Shinji SAITAMA MEDICAL SCHOOL, ANATOMY, ASSOCIATE PROFESSOR, 医学部, 講師 (80129155)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | AMPHIBIA / EMBRYONIC CELL / MAMMALIAN GENE / FORCED EXPRESSION / FUNCTIONAL ANALYSIS / TRIAD / SKELETAL MUSCLE / KNOCKOUT OMUSE / 胚細胞 / 筋小胞体 / T管 / Triad |
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| Research Abstract |
We have exploitated a convenient and quick method for analyzing functions of mammalian genes. In a word, it is a method for presuming gene functions from ultrastructural changes caused in embryonic cells after the gene was forced expressed in the amphibian embryonic cells. We tried the method on two genes (MG29 and junctophilin), which have been isolated from the mammalian skeletal muscle. The results suggested that the MG29 may involve in the formation of networks of sarcoplasmic reticulum, and that junctophilin (if consists of three subtypes, JP-1, JP-2 and JP-3) may involve in the formation of coupling between sarcoplasmic reticulum and transverse tubule (or plasma membrane).
After these experiments using amphibian embryonic cells, knockout mice lacking these genes have been made, and functions of the genes playing for the triad formation in the skeletal muscle cells have been analyzed. The results have shown the following results.
1. Formation of the networks of sarcoplasmic reticulum was abnormal in structure in skeletal muscle of the mutant mice lacking MG29.
2. Formation of coupling between the sarcoplasmic reticulum and cell membrane was impossible in developing cardiac-muscle cells of the mutant mice lacking JP-2.
3. formation of the triads was impossible in skeletal muscle cells of the mutant mice lacking JP-1
In addition, it was also clarified that JP-1 and JP-2 play important roles in the formation of diads and triads respectively in developing skeletal muscle cells. As mentioned above, the method using amphibian embryonic cells was shown to be and easy and effective one for analyzing the functions of mammalian genes.
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Report
(3 results)
Research Products
(18 results)
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[Publications] Ito, K., Komazaki, S. Sasamoto, K., Yoshida, M., Nishi, M., Kitamura, K., and Takeshima, H.: "Deficiency of triad junction and contraction in mutant skeletal muscle lacking junctophilin type 1"J. Cell Biol.. 154. 1059-1067 (2001)
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[Publications] Nishi, Miyuki, Hashimoto, K., Kuriyama, K., komazaki< S., Kano, m., Shibata, S. and Takeshima, H.: "Motor discoordination in mutatnt mice lacking junctophilin type 3"Biochem. Biophys. Res. Comm.. (in press). 2002
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