| Project/Area Number |
17380200
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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 |
Applied molecular and cellular biology
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| Research Institution | Nagoya University |
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Principal Investigator |
MATSUDA Tsukasa Nagoya University, Graduate School of Bioagricultural Sciences, Professor (20144131)
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| Co-Investigator(Kenkyū-buntansha) |
NADANO Daita Nagoya University, Graduate Schcol of Bioagricultural Sciences, Asscoiate Professor (00228074)
SATO Chihiro Nagoya University, Bioscience and Biattchnology Center, Professor (10343211)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,450,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥1,650,000)
Fiscal Year 2007: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
Fiscal Year 2006: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2005: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Fertilization / protein / sugar chains / development / differentiation / applied animal science / 透明体 / Zona Pellucida / 受精 / ZPドメイン |
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| Research Abstract |
From a viewpoint of cell-cell interaction, Structural and functional analyses have been done on two kinds of glycoproteins (ZP glycoprotein of zona pellucid and phospholipid-binding protein, MFG-ES, of sperm surface),which have been suggested to be involved in specific recognition and binding for gamete interaction. As for the ZP glycoprotein, ligand blotting and immuniprecipitation analyses demonstrated that ZPB1 specifically bound to ZPC and thereby disulfide-bridged ZPB1 dimmerization was induced. Such ZPC-induced ZPB1 conformational change was suggested to be responsible for acquisition of sperm activation ability. Furthermore, ZPB1 interacted with ZPC secreted from transfected COS cells and formed insoluble matrixes with ZPC on the cell surface. In vitro incubation of ZPB1 with ZPC spontaneously produced fibrous aggregates of ZPB1-ZPC hetero complexes, which were visible under optical microscopy and morphologically resembled the aggregates obtained from mechanically decomposed chi
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cken egg-envelope. Formation of such fibrous aggregates depended on ZPC/ZPB1 ratio, and involved ZPB1 dimerization through disulfide cross-linking, which had been found in authentic egg-envelope developed in hen's ovary. Furthermore, addition of excessive amounts of ZPC to ZPB1 produced soluble but high molecular weight heterocomplexes with increased adherence property against polystyrene ELISA plates. Thus, the specific association between ZPB1 and ZPC could play pivotal roles to initiate complex formation of hetero-polymers of ZP proteins in egg-envelope matrix construction. As for MFG-E8, this phospholipid-binding protein was shown to be expressed in various types of cells other than sperm by mRNA analyses of various animal cells and tissues, and some important roles in the differentiation and function of adipocytes and the apoptosis of mammary epithelial cells were suggested as a membrane protein on outer surface of microvesicles, termed exosome. Presence of MFG-ES on exosomes suggested contribution of membrane microvesicles to sperm-egg interaction and importance of complex supermolecular interaction among ZP-glycoproteins and the membrane-vesicle constituent molecules, including MFG-E8. Less
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