| Project/Area Number |
10460129
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Nagoya University |
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Principal Investigator |
FUKUTA Katsuhiro Nagoya University, Grad Sch. of Bioagricultural Sciences, Professor, 大学院・生命農学研究科, 教授 (10012022)
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| Co-Investigator(Kenkyū-buntansha) |
HIRUNAGI Kanjun Nagoya University, Museum, Associate Professor, 博物館, 助教授 (00126898)
MATSUDA Yoichi Hokkaido Univ., Center for Adv. Sci. and Technol., Professor, 先端科学技術共同研究センター, 教授 (70165835)
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| Project Period (FY) |
1998 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Tetraploidy / Mouse / Early stage embryo / Suppresion of mitosis / Cytochalasin B / Cell fusion / マウス初期胚 / 細胞分乳抑制 |
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| Research Abstract |
Mitosis prohibition or cell fusion at early developmental stage produces the tetraploid embryos in mammals. However, these embryos can not develop for full term of gestation and never deliver living young. The tetoploid cells survive and are maintained in vitro, but the cells can not form living fetus. The failure of their develop ment are considered to depend on increment of the cell size which results in insufficiency of metabolism and delay of cell cycles. In the present study of the tetraploid mouse embryo induced by cytochalsin B, the delay of tetra ploid cell cycles were confirmed directly in spite of the morula and blastcyst formatidh is performed on time as normal deploid embryos. And continuation of mitosis prohibition produced enormously large nucleus and multi nuclei from 2 to 4 in a cell. For the analysis of the polyploidy, we used the direct R-banding FISH method for the interphase of cell cycle, unlike karyotype analysis at the mitotic phase. By this method the signal of
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BAC clone from MHC class H area was found as 2 single dots in deploid and 4 single dots in tetraploid. It shows that the direct R-banding FISH method is a useful techniques for distinction of chromosomes duplication in interp base. For the trace of the deploid and tetraploid cell during embryogenesis, the chimeric mouse were made using green fluorescent transgenic mouse. Both cell lineages were readily identified under the uv lihgt, as green fluorescent in the GFP-Tg mouse derived cell. As a result of the analysis of chimra between deploid and tetra ploid embryos, tetraploid derived cells composed mainly the extraembryonic tissue, such as placenta and embryonic membrane, in contarast to the deploid derived cells contributed to the embryos. Above mentioned results from the tetraploidy by mitosis inhibition were differ from the tetraploidy by the electric cell fusion. 'The further study should be performed on the difference of tetraploidy between by mitosis inhibition and cell fusion in detail. Less
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