2000 Fiscal Year Annual Research Report

生殖細胞の選択的死滅制御機構の解明とその人為的支配

Research Project

Project/Area Number	11460127
Research Institution	Kyoto University
Principal Investigator	宮本元京都大学, 農学研究科, 教授 (00026618)
Co-Investigator(Kenkyū-buntansha)	杉本実紀京都大学, 農学研究科, 助手 (20243074) 眞鍋昇京都大学, 農学研究科, 助教授 (80243070)
Keywords	アポトーシス / 卵胞 / アポトーシス受容体 / デコイ受容体 / HVJ-リポソーム / 遺伝子導入 / モノクローナル抗体 / ブタ
Research Abstract	サケのように一生にただ一度全卵子を成熟・排卵する動物が存在する一方、哺乳類のように胎児期に減数分裂前期後半の卵核胞期まで進行して休止している卵母細胞が性成熟後順次長い眠りから目覚めて性周期毎に一定数が発育・成熟して排卵されるものが存在する。この過程で卵母細胞の選抜が行われて99%以上が消滅してしまう。卵母細胞と卵胞の分化・発育・成熟は相互に緊密に関連しており、卵母細胞選抜と卵胞選抜は同義の現象と考えられているが、このようにより優秀で強靭な子孫を残す戦略として生殖子の選択的死滅が繰り返されている。本研究の目的は未だ未解明なこの選抜の分子機構を明らかにし、遺伝子レベルでの人為的制御技術を創出することである。そのために卵巣における生殖子の選択的死滅を制御している新規アポトーシス受容体遺伝子の全塩基配列の決定を行い、併せてこの新規アポトーシス受容体遺伝子のHVJ-リポソーム法によるin vivo導入のための実験動物試験を進めて安定導入法を確立している。(1)新規アポトーシス受容体の同定:モノクロナール(PFG-5)抗体を用いて新規アポトーシス受容体を同定した。(2)新規アポトーシス受容体mRNAの同定:健常卵胞の顆粒層細胞の細胞質内でアポトーシス受容体蛋白とリボゾーム複合体を形成しているmRNAをPFG-5抗体を用いて釣り上げ、顆粒層細胞アポトーシス過程におけるアポトーシス受容体遺伝子の発現と局在の推移を明らかとしている。(3)アポトーシスシグナルの細胞内伝達系の分子機構解明:アポトーシス受容体下流にはICE様システインプロテアーゼ(CP)が存在する。CPファミリーはICE系、Apopain系、MHC-1系の3系があるが、申請者の研究から顆粒層細胞には阻害剤プロフィールが何れとも異なる新規CPが存在することが推定されている。このアポトーシスシグナル伝達系の鍵となる新規CPを同定した。

Research Products

(14 results)

All Other

All Publications (14 results)

[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "Anovel in vivo 31P-nuclear magnetic resonance technique for assessment of teratogenicity induced by environmental endocrine disrupters in mice."Japanese Journal of Clinical Medicine. 58・2. 108-111 (2000)
[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "In vivo noninvasive nuclear magnetic resonance technique for assessment of teratogenicity induced by environmental pollutants."Environment Measurement and Chemical Analysis. 27・1. 60-66 (2000)
[Publications] Noboru Manabe,Yasuyoshi Azuma,Hajime Miyamoto et al.: "Effects of the mycelial extract of cultured Cordyceps sinensis on in vivo hepatic energy metabolism and blood flow in dietary hypoferric anemic mice."British Journal of Nutrition. 83・2. 197-204 (2000)
[Publications] Noboru Manabe,Akira Myoumoto,Chiemi Tajima,Hajime Miyamoto et al.: "Immunochemical characteristics of a novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles."Cytotechnology. 33・2. 189-201 (2000)
[Publications] Mizuho Nakayama,Noboru Manabe,Hajime Miyamoto et al.: "Species specific differences in apoptotic cell localization in granulosa and theca interna cells during follicular atresia in porcine and bovine ovaries."Journal of Reproduction and Development . 46・2. 147-156 (2000)
[Publications] Susumu Nishihara,Noboru Manabe,Hajime Miyamoto et al.: "Changes in cell adhesion molecules during follicular atresia in porcine ovaries."Journal of Reproduction and Development. 46・3. 325-334 (2000)
[Publications] Mizuho Nakayama,Noboru Manabe,Hajime Miyamoto et al.: "Changes in localization of type I, III and IV collagens and their mRNA expression during follicular atresia in bovine and porcine ovaries."Journal of Reproduction and Devleopment. 47・1. 27-36 (2001)
[Publications] Kozue Uchio,Noboru Manabe,Kotaro Tamura,Makoto Miyamoto et al.: "Decreased matrix metalloproteinase activity in the kidneys of hereditary nephrotic mice (ICGN strain)."Nephron. 86・1. 145-141 (2000)
[Publications] Kotaro Tamura,Noboru Manabe,Kozue Uchio,Makoto Miyamoto et al.: "Characteristic changes in carbohydrate profile in the kidneys of hereditary nephrotic mice (ICGN strain)."The Journal of Veterinary Medical Science. 52・2. 379-390 (2000)
[Publications] Noboru Manabe,Akiko Kinoshita,,Kozue Yamada-Uchio et al.: "Changes in quantitative profile of extracellular matrix components in the kidneys of rats with adriamycin-induced nephropathy."The Journal of Veterinary Medical Science. 53・1. 125-133 (2001)
[Publications] Miki Sugimoto,Noboru Manabe,Hajime Miyamoto.: "Future functional-morphological studies in embryology."Journal of Reproduction and Development. 46・1. 71-77 (2000)
[Publications] Noboru Manabe,Hajime Miyamoto et al.: "Immunohistochemical quantification of fast-myosin in upper hindlimb muscles of the pig."Animal Science Journal. 72・2. 154-163 (2000)
[Publications] Miki Sugimoto,Noboru Manabe,Hajime Miyamoto.: "Development of infantile rat ovaries autotransplanted after cryopreservation by vitrification."Theriogenology. 53・11. 1093-1103 (2000)
[Publications] Noboru Manabe,Akira Myoumoto,Chiemi Tajima,Hajime Miyamoto et al.: "A novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles."Tohoku University Press, Sendai, Japan.. 357 (2000)

2000 Fiscal Year Annual Research Report

生殖細胞の選択的死滅制御機構の解明とその人為的支配

Principal Investigator

宮本 元 京都大学, 農学研究科, 教授 (00026618)

Research Products

[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "Anovel in vivo 31P-nuclear magnetic resonance technique for assessment of teratogenicity induced by environmental endocrine disrupters in mice."Japanese Journal of Clinical Medicine. 58・2. 108-111 (2000)

[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "In vivo noninvasive nuclear magnetic resonance technique for assessment of teratogenicity induced by environmental pollutants."Environment Measurement and Chemical Analysis. 27・1. 60-66 (2000)

[Publications] Noboru Manabe,Yasuyoshi Azuma,Hajime Miyamoto et al.: "Effects of the mycelial extract of cultured Cordyceps sinensis on in vivo hepatic energy metabolism and blood flow in dietary hypoferric anemic mice."British Journal of Nutrition. 83・2. 197-204 (2000)

[Publications] Noboru Manabe,Akira Myoumoto,Chiemi Tajima,Hajime Miyamoto et al.: "Immunochemical characteristics of a novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles."Cytotechnology. 33・2. 189-201 (2000)

[Publications] Mizuho Nakayama,Noboru Manabe,Hajime Miyamoto et al.: "Species specific differences in apoptotic cell localization in granulosa and theca interna cells during follicular atresia in porcine and bovine ovaries."Journal of Reproduction and Development . 46・2. 147-156 (2000)

[Publications] Susumu Nishihara,Noboru Manabe,Hajime Miyamoto et al.: "Changes in cell adhesion molecules during follicular atresia in porcine ovaries."Journal of Reproduction and Development. 46・3. 325-334 (2000)

[Publications] Mizuho Nakayama,Noboru Manabe,Hajime Miyamoto et al.: "Changes in localization of type I, III and IV collagens and their mRNA expression during follicular atresia in bovine and porcine ovaries."Journal of Reproduction and Devleopment. 47・1. 27-36 (2001)

[Publications] Kozue Uchio,Noboru Manabe,Kotaro Tamura,Makoto Miyamoto et al.: "Decreased matrix metalloproteinase activity in the kidneys of hereditary nephrotic mice (ICGN strain)."Nephron. 86・1. 145-141 (2000)

[Publications] Kotaro Tamura,Noboru Manabe,Kozue Uchio,Makoto Miyamoto et al.: "Characteristic changes in carbohydrate profile in the kidneys of hereditary nephrotic mice (ICGN strain)."The Journal of Veterinary Medical Science. 52・2. 379-390 (2000)

[Publications] Noboru Manabe,Akiko Kinoshita,,Kozue Yamada-Uchio et al.: "Changes in quantitative profile of extracellular matrix components in the kidneys of rats with adriamycin-induced nephropathy."The Journal of Veterinary Medical Science. 53・1. 125-133 (2001)

[Publications] Miki Sugimoto,Noboru Manabe,Hajime Miyamoto.: "Future functional-morphological studies in embryology."Journal of Reproduction and Development. 46・1. 71-77 (2000)

[Publications] Noboru Manabe,Hajime Miyamoto et al.: "Immunohistochemical quantification of fast-myosin in upper hindlimb muscles of the pig."Animal Science Journal. 72・2. 154-163 (2000)

[Publications] Miki Sugimoto,Noboru Manabe,Hajime Miyamoto.: "Development of infantile rat ovaries autotransplanted after cryopreservation by vitrification."Theriogenology. 53・11. 1093-1103 (2000)

[Publications] Noboru Manabe,Akira Myoumoto,Chiemi Tajima,Hajime Miyamoto et al.: "A novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles."Tohoku University Press, Sendai, Japan.. 357 (2000)

宮本元京都大学, 農学研究科, 教授 (00026618)