| Project/Area Number |
09460127
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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 | KYOTO UNIVERSITY |
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Principal Investigator |
MANANBE Noboru Kyoto Univ., Dpt. Agriculture, Associate Professor, 農学研究科, 助教授 (80243070)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUMOTO Manabu Tohoku Univ., Dpt. Medicine, Professor, 加齢医学研究所, 教授 (60156809)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥13,600,000 (Direct Cost: ¥13,600,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1997: ¥7,400,000 (Direct Cost: ¥7,400,000)
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| Keywords | Ovary / Follicle / Granulosa cell / Apoptosis / Cell death receptor / Porcine / Monoclonal antibody / Programmed cell death / 卵胞顆粒層細胞 / 細胞内シグナル伝達系 / mRNA / ブタ卵巣 / カスパーゼ / 分子制御機構 |
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| Research Abstract |
In mammalian ovaries, more than 99% of the follicles undergo the degenerative change known as "atresia" at varying stages of follicle development, and a number of studies of follicular atresia have revealed the morphological and biochemical characteristics of atretic follicles. Recent findings have suggested that granulosa cell apoptosis is the mechanism underlying ovarian follicular atresia. However, regulation mechanisms of ovarian cell apoptosis are not well understood. l produced unique monoclonal antibodies to reveal the molecular mechanisms regulating follicular granulosa cell apoptosis in porcine ovaries. lgM monoclonal antibodies (PFG-1 and -3) capable of inducing granulosa cell apoptosis and an lgG monoclonal antibody (PFG-4) not capable of inducing apoptosis were produced against granulosa cells prepared from antral follicles of porcine ovaries. Two-dimensional Western blotting analysis revealed that PFG 1 specifically recognized a cell-membrane protein (pl 5.9; named PFG-1 a
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ntigen) of granulosa cells of healthy follicles, and that PFG-3 recognized two cell-membrane proteins (42kD, pl 6.2 and 55 kD, pl 5.9; named PFG-3 and PFG-1 antigens, respectively) of granulosa cells of healthy follicles and PFG-4 recognized the same two membrane proteins. In granulosa cells of atretic follicles, PFG-1 antigen disappeared and only PFG-3 antigen was immunohistochemically detected. Immunochemical reactions against these antibodies were only detected in follicular granulosa cells but not any other ovarian tissues or other organs. PFG-1 immunohistochemically reacted with granulosa cells of healthy follicles, and PFG-3 and PFG-4 reacted with granulosa cells of both healthy and atretic follicles. When the isolated granulosa cells prepared from healthy follicles were cultured in medium containing PFG-1 or -3 (more than 0.001 μg/ml); the cells underwent apoptosis, and co-incubation with PFG-4 inhibited the antibody-inducible apoptosis. These present observations suggested that PFG-1 antigen is a novel cell death receptor which is different from the apoptosis-mediating receptors(Fas or TNF receptor), and that PFG-3 antigen may act as a decoy receptor and inhibit apoptotic signal induced by lgM antibodies. Less
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