| Project/Area Number |
14370306
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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 |
Hematology
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| Research Institution | Keio University |
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Principal Investigator |
OHBO Kazuyuki Keio University, Department of Medicine, Assistant professor, 医学部, 講師 (70250751)
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| Co-Investigator(Kenkyū-buntansha) |
SUDA Toshio Keio University, School of Medicine, Professor, 医学部, 教授 (60118453)
YOSHIDA Shosei Kyoto University, School of Medicine, Assistant professor, 大学院・医学研究科, 助手 (60294138)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥8,700,000 (Direct Cost: ¥8,700,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2002: ¥4,300,000 (Direct Cost: ¥4,300,000)
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| Keywords | Cell and Tissue / Regenerative medicine / Development and Differentiation / Anatomy / Signal transduction / 生殖幹細胸 / 造血幹細胞 / 細胞表面蛋白 / ニッチ / 生殖幹細胞 / ニッシェ |
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| Research Abstract |
The common properties such as self-renewing and necessity of supporting cells called niche are maintained by either identical or similar molecular mechanisms. Among those common properties, we are interested in interacting molecules between stem cells and niches. Therefore, we decided to focus on cell surface molecules and secreting molecules that are crucial for interaction between stem cells and supporting cells.
We generated cDNA library from purified male germ stem cells of Oct-4/GFP mice that carry a transgene expressing EGFP protein under the control of Oct-4 promoter and enhancer. We cloned genes encoding cell surface molecules and secreting molecules by a signal sequence trap method. Then, we chose the molecules that were not expressed in somatic cells, Serotoli cells and NIH3T3 fibroblasts. As consequence, we decided to focus on two molecules, mHE-4 and JAM-4. mHE-4 is one of WAP type serine proteinase inhibitor proteins and JAM-4 is a immunoglobulin type cell-adhesion molecule
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. Then, we observe the expression of those molecules by RT-PCR, northen blotting analysis and immunohistochemical analysis. Both of them were expressed in CD34-negative hematopoietic stem cells. CD34-positive hematopoietic stem cells expressed both molecules but weakly. The progenitor cells express neither mHE-4 nor JAM-4. In addition, immunohistochemical analysis showed JAM-4 was expressed in liver and kidney, and HE-4 was expressed in lung and kidney. Proteinases and their inhibitors play roles at many biological reactions. Because serine proteinase functions in digestion of bacreia, mHE may function at the process of bacterial infection. In testis, mHE-4 expressed on the cell surface of spermatogonia. Histochemical analysis of JAM-4 showed the co-localization with tight junction protein, ZO-1, suggesting JAM-4 may function as a tight junction protein. To analyze the function of those molecules in vivo, we are currently generating knock-out mice of those molecules and have already obtained the chimera mice. Less
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