2002 Fiscal Year Annual Research Report

内分泌攪乱物質による生殖毒性評価システムの構築:核磁気共鳴イメージング法による構造生物学的リアルタイム評価法の確立

Research Project

Project/Area Number	13027241
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)	宮川恒京都大学, 農学研究科, 教授 (10219735) 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) 奥田潔岡山大学, 自然科学研究科, 教授 (40177168) 木曾康郎山口大学, 農学部, 教授 (10142374)
Keywords	外因性内分泌攪乱物質 / 核磁気共鳴イメージング / 三次元形態解析 / 哺乳類 / 胎仔 / オーファン受容体 / 受容体ノックアウト / リアルタイム解析
Research Abstract	前世紀末、産業革命期以後200年にわたって人類が創り出してきた農薬、医薬、食品添加物等の多くが、生体の内分泌系のシグナル伝達を攪乱する内分泌攪乱物質として人類の生存を脅かすことが分かってきた。本研究は、生殖生理学の基盤に関わる新技術開発や生殖子の選抜機構研究を進め、新規な原理に基づいた非侵襲的かつリアルタイムに生殖毒性を評価できる技術システムを構築し、従来法とは比較できない精度で速やかに生殖毒性を評価できる先端技術を確立することで、様々な化合物の生殖毒性を的確に評価できる未来を開拓する基盤技術を開発するものである。本年度は、構造生物学、細胞生物学的視点から内分泌攪乱物質とホルモン受容体の結合様式、受容体分子の立体構造変化等を解析し、環境系に存在する様々な化合物の生殖毒性をリアルタイムに評価するシステムの確立を下記のように進めた。麻酔下の動物に生体NMR(NMRシグナルを定量検出できる特殊プローブの開発を進めた)に保定し、胎児におけるNMRシグナルを部位特異的にリアルタイムに検出し、3次元立体画像データとして解析できる測定アプリケーションの作成と最適測定条件の決定を行った。毒性発現機構の生殖生理学的解明のため、遺伝子工学的にオーファン受容体に様々な構造変異を誘導し、化合物と受容体の相互的結合特性をNMRにて観測して分子構造学的に毒性を予測する技術を開発している。卵母細胞は遺伝子に制御された細胞死によって選択されているが、これを制御する新規細胞死受容体(申請者らが発見した卵胞顆粒層細胞特異的アポトーシス受容体)を介した細胞死シグナル伝達機構を解明し、化合物の影響を分子レベルで評価する系の開発を進めた。雄では、精巣セルトリ細胞のプロラクチン-インヒビンB受容体複合体が精子形成過程におけるアポトーシスを支配的に制御していることを見出したので、この分子機構の解明を進めている。

Research Products
(24 results)

All Other

All Publications (24 results)

[Publications] Mizuho Nakayama, Noboru Manabe, Naoko Inoue, Toshikatsu Matsui, Hajime Miyamoto: "Changes in the expression of tumor necrosis factor (TNF) a, TNFa Receptor (TNFR) 2 and TNFR-associated factor 2 in granulosa cells during atresia in pig ovaries"Biology of Reproduction. 68・2. 530-535 (2003)
[Publications] Uchio K, Tuchwdber B, Manabe Noboru, Gabbiani G, Rosenbaum J, Desmouliere A: "Cellular retinol-binding protein-1 expression and modulation during in vivo and in vitro myofibroblastic differentiation of rat hepatic stellate cells and portal fibroblasts"Laboratory Investigation. 82・4. 619-628 (2002)
[Publications] Chiemi Tajima, Noboru Manabe, Mizuho Nakayama, Naoko Inoue, Toshikatsu Matsui, Eiichi Hondo, Hajime Miyamoto Hajime: "Monoclonal antibody recognizes follicular granulosa cell antigens in porcine ovaries"Journal of Reproduction and Devevelopment. 48・4. 567-572 (2002)
[Publications] Satoshi Imanishi, Noboru Manabe, Takako Tanaka, Reika Okamoto, Maki Morita, Miki Sugimoto, Hajime Miyamoto: "mRNA of GPCR5B/Raig2, a novel orphan receptor, expresses in murine placentae"Journal of Reproduction and Devevelopment. 48・4. 627-632 (2002)
[Publications] Noboru Manabe: "Molecular mechanism of follicle selection in mammalian ovaries"Journal of Laboratory Animals (Kansai). 23・1. 71-78 (2002)
[Publications] Noboru Manabe, Miki Sugimoto, Hajime Miyamoto: "In vivo noninvasive nuclear magnetic resonance (NMR) technique for assessment of teratogenicity induced by chemicals"Chemical Engineering. 47・2. 110-114 (2002)
[Publications] Satoko Wada, Noboru Manabe, Naoko Inoue, Mizuho Nakayama, Toshikatsu Matsui, Hajime Miyamoto: "TRAIL-decoy receptor-1 disappears in granulosa cells of atretic follicles in porcine ovaries"Journal of Reproduction and Devevelopment. 48・1. 167-173 (2002)
[Publications] Satoko Wada, Noboru Manabe, Naoko Inoue, Mizuho Nakayama, Toshikatsu Matsui, Hajime Miyamoto: "TRADD is involved in apoptosis induction in granulosa cells during atresia in pig ovaries"Journal of Reproduction and Devevelopment. 48・1. 175-181 (2002)
[Publications] Hideki Tonomura H, Miyake T, Taniguchi Y, Manabe Noboru, Kose H, Matsumoto K, Yamada T, Sasaki Y: "Detection of quantitative trait locus for rat model useful for genetic analysis of intramascular fat accumulation using the OLETF rat"The Journal of Veterinary Medical Science. 64・1. 45-50 (2002)
[Publications] Satoko Wada, Noboru Manabe, Mizuho Nakayama, Naoko Inoue, Toshikatsu Matsui, Hajime Miyamoto: "TRAIL-decoy receptor-1 plays inhibitory role in apoptosis of granulosa cells from pig ovarian follicules"The Journal of Veterinary Medical Science. 64・5. 435-439 (2002)
[Publications] Miki Sugimoto, Noboru Manabe, Maki Morita, Takako Tanaka, Reika Okamoto, Satoshi Imanishi, Hajime Miyamoto: "NMR microscopic observation of mouse embryo malformations induced by maternal administration of retinoic acid"The Journal of Veterinary Medical Science. 64・5. 427-433 (2002)
[Publications] Miki Sugimoto, Noboru Manabe, Hajime Miyamoto: "Cryopreservation techniques for ovarian tissues"Hormone Frontier in Gynecology. 9・1. 135-140 (2002)
[Publications] Mizuho Nakayama, Noboru Manabe, Naoko Inoue, Toshikatsu Matsui, Shinsuke Nakagawa, Akihisa Maeda, Hajime Miyamoto Sakamaki, Miki Sugimoto, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Tumor necrosis factor-α acts as a survival factor in granulosa cells during follicular selection in porcine ovaries. PP.143-149"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Naoko Inoue, Noboru Manabe, Toshikatsu Matsui, Shinsuke Nakagawa, Akihisa Maeda, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Roles of Fas-ligand and TRAIL systems in granulosa cell apoptosis during follicular atresia in porcine ovaries. PP.151-156"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Toshikatu Matsui, Noboru Manabe, Yasufumi Goto, Naoko Inoue, Shinsuke Nakagawa, Akihisa Maeda, Susumu Nishihara, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Role of the mitochondria-dependent pathway in granulosa cell apoptosis during follicular atresia in pig ovaries. PP.157-163"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Yamaguchi-Yamada M, Noboru Manabe, Uchio-Yamada K, Akashi N, Gotoh Y, Nagao M, Yamamoto Y, Ogura A, Miyamoto H.: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Anemia associated with renal failure in the ICR-derived glomerulonephritis (ICGN) mice. PP.281-285"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Minako Koso, Noboru Manabe, Kohji Komatsu, Munetake Shimabe, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Abnormal structural luteolysis in ovaries of the senescence accelerated mouse (SAM) : expression_of Fas mediated apoptosis signal in luteal bodies. PP.287-294"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Noboru Manabe, Kozue Yamada-Uchio, Misuzu Yamada-Yamaguchi, Yasufumi Gotoh, Toshiharu Mori, Chinatsu Sakata, Yoshihiro Furuya, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Histochemical technique quantitatively estimates the physiological and pathological properties. PP.303-309"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Noboru Manabe, Naoko Inoue, Toshikatsu Matsui, Shinsuke Nakagawa, Akihisa Maeda, Takashi Miyano, Kazuhiro Sakamaki, Miki Sugimoto Hajime: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Regulatory mechanisms of follicle selection in mammalian ovaries. PP.117-132"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Noboru Manabe, Naoko Inoue, Toshikatsu Matsui, Shinsuke Nakagawa, Akihisa Maeda, Takashi Miyano, Kazuhiro Sakamaki, Miki Sugimoto, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Tumor necrosis factor-related apoptosis-inducing ligand and its receptors play crucial roles in apoptosis of granulosa cells during follicular atresia in pig ovaries. PP.133-142"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Maki Morita, Miki Sugimoto, Noboru Manabe, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Morphological alterations in mammalian reproductive organ induced by exposure to diethyistilbestrol (DES). PP.319-323"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Miki SUGIMOTO, Noboru MANABE, Maki MORITA, Hajime MIYAMOTO: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Utilization of magnetic resonance microscope for studies in reproductive toxicology. PP.325-330"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Noboru Manabe, Yasuyoshi Azuma, Miki Sugimoto, Kei-Ichi Sezaki, Mineko Iwahori, Satoshi Imanishi, Hanako Nishizawa, Takako Tanaka, Maki Morita, Hajime Miyamoto: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), In vivo and real time estimation of energy metabolism by ^<31>p-nuclear magnetic resonance. PP.339-347"Hokuto Publication Company, Kyoto, Japan. 363 (2003)
[Publications] Morikawa Masahiro, Wakoto Matsuyama, Yuko Kawamoto, Miki Sugimoto, Maki Morita, Aki Nakayama, Noboru Mabane, Minoru Yoneda, Shinsuke Morisawa: "Environmental Toxicity Evaluation and Risk Management Vol.1 (Morisawa S eds.), Dynamic performance of bisphenol-A (BPA) in pregnant mouse and physiologically-based pharmacokinetic model development. PP.60-74"The Environment Risk Management Group of The JSPS-VCC Program on Environmental Science, Engineering and Ethics 2000-2009. Kyoto, Japan. 159 (2002)

2002 Fiscal Year Annual Research Report

内分泌攪乱物質による生殖毒性評価システムの構築:核磁気共鳴イメージング法による構造生物学的リアルタイム評価法の確立

Principal Investigator

宮本 元 京都大学, 農学研究科, 教授 (00026618)

Research Products

[Publications] Chiemi Tajima, Noboru Manabe, Mizuho Nakayama, Naoko Inoue, Toshikatsu Matsui, Eiichi Hondo, Hajime Miyamoto Hajime: "Monoclonal antibody recognizes follicular granulosa cell antigens in porcine ovaries"Journal of Reproduction and Devevelopment. 48・4. 567-572 (2002)

[Publications] Satoshi Imanishi, Noboru Manabe, Takako Tanaka, Reika Okamoto, Maki Morita, Miki Sugimoto, Hajime Miyamoto: "mRNA of GPCR5B/Raig2, a novel orphan receptor, expresses in murine placentae"Journal of Reproduction and Devevelopment. 48・4. 627-632 (2002)

[Publications] Noboru Manabe: "Molecular mechanism of follicle selection in mammalian ovaries"Journal of Laboratory Animals (Kansai). 23・1. 71-78 (2002)

[Publications] Noboru Manabe, Miki Sugimoto, Hajime Miyamoto: "In vivo noninvasive nuclear magnetic resonance (NMR) technique for assessment of teratogenicity induced by chemicals"Chemical Engineering. 47・2. 110-114 (2002)

[Publications] Satoko Wada, Noboru Manabe, Naoko Inoue, Mizuho Nakayama, Toshikatsu Matsui, Hajime Miyamoto: "TRAIL-decoy receptor-1 disappears in granulosa cells of atretic follicles in porcine ovaries"Journal of Reproduction and Devevelopment. 48・1. 167-173 (2002)

[Publications] Satoko Wada, Noboru Manabe, Naoko Inoue, Mizuho Nakayama, Toshikatsu Matsui, Hajime Miyamoto: "TRADD is involved in apoptosis induction in granulosa cells during atresia in pig ovaries"Journal of Reproduction and Devevelopment. 48・1. 175-181 (2002)

[Publications] Satoko Wada, Noboru Manabe, Mizuho Nakayama, Naoko Inoue, Toshikatsu Matsui, Hajime Miyamoto: "TRAIL-decoy receptor-1 plays inhibitory role in apoptosis of granulosa cells from pig ovarian follicules"The Journal of Veterinary Medical Science. 64・5. 435-439 (2002)

[Publications] Miki Sugimoto, Noboru Manabe, Maki Morita, Takako Tanaka, Reika Okamoto, Satoshi Imanishi, Hajime Miyamoto: "NMR microscopic observation of mouse embryo malformations induced by maternal administration of retinoic acid"The Journal of Veterinary Medical Science. 64・5. 427-433 (2002)

[Publications] Miki Sugimoto, Noboru Manabe, Hajime Miyamoto: "Cryopreservation techniques for ovarian tissues"Hormone Frontier in Gynecology. 9・1. 135-140 (2002)

[Publications] Miki SUGIMOTO, Noboru MANABE, Maki MORITA, Hajime MIYAMOTO: "Animal Frontier Science (Sato E, Miyamoto H, Manabe Noboru eds.), Utilization of magnetic resonance microscope for studies in reproductive toxicology. PP.325-330"Hokuto Publication Company, Kyoto, Japan. 363 (2003)

宮本元京都大学, 農学研究科, 教授 (00026618)