2000 Fiscal Year Annual Research Report

選択的卵胞閉鎖の分子制御機構の解明とその人為支配

Research Project

Project/Area Number	12460125
Research Institution	Kyoto University
Principal Investigator	眞鍋昇京都大学, 農学研究科, 助教授 (80243070)
Co-Investigator(Kenkyū-buntansha)	杉本実紀京都大学, 農学研究科, 助手 (20243074)
Keywords	アポトーシス / 卵胞 / 細胞死受容体 / 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	哺乳類の片側卵巣には胎児期に減数分裂前期後半で休止した卵母細胞が5〜10万個含まれている。性周期毎に繰り返される排卵の過程で99%以上が消滅してしまい、1%以下が排卵に至る。この選抜の結果、優秀で強靭な子孫が残ると考えられるが、その制御機構は未解明である。本研究の目的は、選抜の分子機構を明らかにし、ex vovo遺伝子導入によって人為支配技術を創出することにある。そのために(1)選択的卵胞死滅の制御機構の解明:既知細胞死受容体と異なる卵胞顆粒層細胞に特異的な新規アポトーシス受容体を見出しており、これを介した卵胞の選択的死滅の分子制御機構を明らかとする。次いで(2)ex vovo遺伝子導入による細胞死の人為支配技術の創出:細胞死受容体およびこのシグナル伝達系に関わるカスパーゼをコードする遺伝子を、HVJ-リポソーム法にて卵巣にex vovo導入し、卵胞の選択的死滅の人為支配技術を創出する。今年度は(1)新規アポトーシス受容体の同定とシグナル伝達系の解析:健常卵胞から調製した初代培養顆粒層細胞にアポトーシスを特異的に誘導できるモノクローナル抗体が認識する細胞膜結合性糖蛋白は、既知アポトーシス受容体とは異なる新規受容体である。この抗体を用いてアフィニティーカラムを作製し、アポトーシス受容体を精製してアミノ酸配列を決定している。併せて、特異的mRNAをアフィニティーカラムにて調製し、RT-DNAシーケンス法にて全塩基配列を決定している。予備研究から、新規アポトーシス受容体には既知受容体と同様に「death-domain」が存在するので、RT-PCR検出系を作成するとともに、局在解析のためにin situ hybridizationプローブを作成して、顆粒層細胞のアポトーシス過程および黄体細胞化への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た。(2)in vivo遺伝子導入法の確立:卵胞閉鎖を人為支配するため、SABレクチン/HVJ-リポソーム遺伝子導入法の基礎検討を行った。

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "A novel 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 Development. 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 hind limb 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 granulos a cells of porcine ovarian follicles."Tohoku University Press,Sendai,Japan.. 357 (2000)

2000 Fiscal Year Annual Research Report

選択的卵胞閉鎖の分子制御機構の解明とその人為支配

Principal Investigator

眞鍋 昇 京都大学, 農学研究科, 助教授 (80243070)

Research Products

[Publications] Noboru Manabe,Miki Sugimoto,Maki Morita,Hajime Miyamoto et al.: "A novel 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 Development. 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 hind limb 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 granulos a cells of porcine ovarian follicles."Tohoku University Press,Sendai,Japan.. 357 (2000)

眞鍋昇京都大学, 農学研究科, 助教授 (80243070)