| Project/Area Number |
15380191
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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 animal science
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| Research Institution | Shizuoka University |
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Principal Investigator |
MORI Makoto Shizuoka University, Faculty of Agriculture, Professor, 農学部, 教授 (90143411)
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| Co-Investigator(Kenkyū-buntansha) |
SASANAMI Tomohiro Shizuoka University, Faculty of Agriculture, Instructor, 農学部, 助手 (80322139)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥15,700,000 (Direct Cost: ¥15,700,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2004: ¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 2003: ¥8,200,000 (Direct Cost: ¥8,200,000)
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| Keywords | Vitelline membrane / Oocyte / Quail / Domestic fowl / Egg coat / Gene / Extracellular matrix / Aves / ZPC / ZP1 / 卵巣 / タンパク分泌 / 繊維形成 / 透明帯 / 卵胞 / 肝臓 / CHO細胞 |
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| Research Abstract |
1.The extracellular matrix surrounding the oocyte before ovulation is called perivitelline membrane in avian species. The perivitelline membrane is constructed with two major glycoproteins, ZPC and ZP1 that are synthesized in the ovarian granulosa cells and the liver, respectively. Although the properties of the major components of the perivitelline membrane had been examined, the knowledge of the nature of fibril formation and secretion pattern from the cell is lacking. In this study, we focused on the binding of quail ZPC and quail ZP1, and comparison of gene structure of various avian species. Moreover, the hypothesis that ZPC binding protein shows an important role for the fibril formation in immature follicle was examined.
2.The microsomal fraction of the granulosa cell was immunoblotted with anti-ZPC after SDS-polyacrylamido gel electrophoresis, and a novel protein migrated at 200 kDa was detected. This band was not reacted with anti-ZPC antiserum or anti-ZP1 antiserum, but strongly bind to ZPC. These results indicated the existence of ZPC binding protein for the fibril formation of perivitelline membrane.
3.In order to compare the binding affinity of quail ZPC to ZP1 of the various avian species, the gel electrophoresis analysis was performed and found that quail ZPC can bind to other partidge ZP1. Turkey and duck ZP1 was cloned their nucleic acid sequences were determined. The results showed that the protein has variance at C terminal amino acids. Moreover, mutant ZP1 was expressed in CHO-K1 cells, and it is bind to ZPC strongly. These results indicated that the N-terminal proteolytic processing of ZP1 might take place after the arrival of ZP1 at the ovary, and the resulting product is incorporated into the perivitelline membrane in species-specific manner.
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