実験用小型淡水魚類卵子の凍結保存:耐凍剤チャンネルの人為的発現によるアプローチ

2009 年度 研究成果報告書

• 研究課題/領域番号: 19300147
• 研究種目: 基盤研究(B)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 実験動物学
• 研究機関: 高知大学
• 研究代表者: 葛西 孫三郎 高知大学, 教育研究部自然科学系, 教授 (60152617)
• 研究分担者: 枝重 圭祐 高知大学, 教育研究部自然科学系, 教授 (30175228) ; 越本 知大 宮崎大学, フロンティア科学実験総合センター, 教授 (70295210)
• 研究期間 (年度): 2007 – 2009
• キーワード: 凍結保存 / 魚類卵子 / 水チャンネル / ゼブラフィッシュ / 細胞膜透過性

研究概要

魚類卵子を凍結保存するには、サイズが小さくて細胞膜透過性が高い、未熟な未成熟卵子が適している。まず、ゼブラフィッシュのStage III中期卵子の体外成熟培養系を開発した。次いで、Stage III中期卵子に一時的に水・耐凍剤チャンネル(aquaporin 3)を発現させると、成熟率～孵化率を低下させずに細胞膜透過性を向上できることを明らかにした。しかし、卵子のガラス化凍結保存には成功しなかった。

研究成果

• [雑誌論文] Equilibrium vitrification of mouse embryos (2010)
  • 著者名/発表者名: Bo Jin, Keiji Mochida, Atsuo Ogura, Eri Hotta, Yukiko Kobayashi, Kaori Ito, Go Egawa, Shinsuke Seki, Hiroshi Honda, Keisuke Edashige, Magosaburo Kasai
  • 雑誌名: Biology of Reproduction 82(2) ページ: 444-450
  • 査読あり
• [雑誌論文] Development of a reliable in vitro maturation system for zebrafish oocytes (2008)
  • 著者名/発表者名: Shinsuke Seki, Toshimitsu Kouya, Ryoma Tsuchiya, Delgado M. Valdez Jr., Bo Jin, Takao Hara, Naoya Saida, Magosaburo Kasai, Keisuke Edashige
  • 雑誌名: Reproduction 135(3) ページ: 285-292
  • 査読あり
• [雑誌論文] Formation of extracellular and intracellular ice during warming of vitrified mouse morulae and its effect on embryo survival (2008)
  • 著者名/発表者名: Bo Jin, Kenji Kusanagi, Makiko Ueda, Shinsuke Seki, Delgado M. Valdez Jr., Keisuke Edashige, Magosaburo Kasai
  • 雑誌名: Cryobiology 56(3) ページ: 233-240
  • 査読あり
• [雑誌論文] The mechanism by which mouse spermatozoa are injured during freezing (2008)
  • 著者名/発表者名: Bo Jin, Chihiro Yamasaki, Naoko Yamada, Shinsuke Seki, Delgado M. Valdez Jr., Magosaburo Kasai, Keisuke Edashige
  • 雑誌名: Journal of Reproduction and Development 54(4) ページ: 265-269
  • 査読あり
• [雑誌論文] The permeability to water and cryoprotectants of immature and mature oocytes in the zebrafish (Danio rerio) (2007)
  • 著者名/発表者名: Shinsuke Seki, Toshimitsu Kouya, Delgado M. Valdez Jr., Bo Jin, Takao Hara, Naoya Saida, Magosaburo Kasai, Keisuke Edashige
  • 雑誌名: Cryobiology 54(1) ページ: 121-124
  • 査読あり
• [雑誌論文] Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio) (2007)
  • 著者名/発表者名: Shinsuke Seki, Toshimitsu Kouya, Takao Hara, Delgado M. Valdez Jr., Bo Jin, Magosaburo Kasai, Keisuke Edashige
  • 雑誌名: Journal of Reproduction and Development 53(3) ページ: 597-603
  • 査読あり
• [雑誌論文] The role of aquaporin 3 in the movement of water and cryoprotectants in mouse morulae (2007)
  • 著者名/発表者名: Keisuke Edashige, Satoshi Ohta, Mitsunobu Tanaka, Tatsunaga Kuwano, Delgado M. Valdez Jr., Takao Hara, Bo Jin, Sei-ichi Takahashi, Shinsuke Seki, Chihiro Koshimoto, Magosaburo Kasai
  • 雑誌名: Biology of Reproduction 77(2) ページ: 365-375
  • 査読あり
• [学会発表] Equilibrium vitrification of mouse embryos (2009)
  • 著者名/発表者名: Bo Jin, Eri Hotta, Yukiko Kobayashi, Kaori Ito, Go Egawa, Shinsuke Seki, Hiroshi Honda, Keiji Mochida, Atsuo Ogura, Keisuke Edashige, Magosaburo Kasai
  • 学会等名: The 46th Annual Meeting of the Society for Cryobiology
  • 発表場所: Sapporo
  • 年月日: 20090719-20090723
• [学会発表] Survival of oocytes in antral follicles of vitrified mouse ovaries (2009)
  • 著者名/発表者名: Keisuke Edashige, Delgado M. Valdez Jr., Hiroshi Honda, Yu Nishikado, Bo Jin, Magosaburo Kasai
  • 学会等名: The 46th Annual Meeting of the Society for Cryobiology
  • 発表場所: Sapporo
  • 年月日: 20090719-20090723
• [学会発表] Rapid movement of water and cryoprotectants in pig expanded blastocysts via channel processes-its relevance to their tolerance to cryopreservation- (2009)
  • 著者名/発表者名: Keisuke Edashige, Bo Jin, Ryu-ichi Higashiyama, Jun-ichi Yonezawa, Masashi Miyake, Sei-ichi Takahashi, Ken-ichi Yazawa, Magosaburo Kasai
  • 学会等名: The 46th Annual Meeting of the Society for Cryobiology
  • 発表場所: Sapporo
  • 年月日: 20090719-20090723
• [学会発表] Effect of the expression of aquaporins 1 and 3 in mouse MII oocytes on the nucleation temperature for intracellular ice formation (2009)
  • 著者名/発表者名: Shinsuke Seki, Keisuke Edashige, Peter Mazur
  • 学会等名: The 46th Annual Meeting of the Society for Cryobiology
  • 発表場所: Sapporo
  • 年月日: 20090719-20090723
• [学会発表] Magosaburo Kasai, Keisuke Edashige. Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio) (2008)
  • 著者名/発表者名: Shinsuke Seki, Bo Jin, Tsuchiya Ryoma, Toshimitsu Kouya, Takao Hara, Delgado M. Valdez Jr
  • 学会等名: The 5th Annual Conference of the Asia Reproductive Biotechnology Society
  • 発表場所: Kunming
  • 年月日: 20081127-20081201
• [学会発表] Permeability to water and cryoprotectants of zebrafish (Danio Rerio) oocytes at mid stage III (2008)
  • 著者名/発表者名: Delgado M. Valdez. Jr., Ryoma Tsuchiya, Shinsuke Seki, Naoya Saida, Bo Jin, Masahiro Yoshimura, Chihiro Koshimoto, Magosaburo Kasai, Keisuke Edashige
  • 学会等名: The 45th Annual Meeting of the Society for Cryobiology
  • 発表場所: Charlotte
  • 年月日: 20080720-20080723
• [学会発表] The pathway for the movement of water and cryoprotectants in bovine oocytes and embryos (2008)
  • 著者名/発表者名: Keisuke Edashige, Takao Hara, Yasunori Kawai, Masahiro Yoshimura, Bo Jin, Delgado M. Valdez, Jr., Magosaburo Kasai
  • 学会等名: The 45th Annual Meeting of the Society for Cryobiology
  • 発表場所: Charlotte
  • 年月日: 20080720-20080723
• [学会発表] Effects of raffinose concentration and methyl-beta-cyclodextrin on IVF with cryopreserved C57BL/6 sperm (2008)
  • 著者名/発表者名: Keiji Mochida, Chihiro Koshimoto, Keisuke Edashige, Kentaro Aizawa, Kyuichi Taguma, Akihiko Ohta, Atsuo Ogura
  • 学会等名: The 45th Annual Meeting of the Society for Cryobiology
  • 発表場所: Charlotte
  • 年月日: 20080720-20080723
• [学会発表] Simple technique to improve motility and fertility in frozen C57BL/6J mouse sperm (2007)
  • 著者名/発表者名: Chihiro Koshimoto, Yuuki Schichi, Keisuke Edashige, Magosaburo Kasai, Keiji Mochida, Akio Shinohara
  • 学会等名: The 44th annual Meeting of the Society for Cryobiology
  • 発表場所: Lake Louse
  • 年月日: 20070828-20070801
• [学会発表] ryobiological properties of immature oocytes assessed by the ability to be fertilized and to develop to term (2007)
  • 著者名/発表者名: Shinsuke Seki, Kouya Toshimitsu, Ryoma Tsuchiya, Delgado M Valdez Jr., Bo Jin, Naoya Saida, Magosaburo Kasai, Keisuke Edashige
  • 学会等名: The 44th Annua Meeting of the Society for Cryobiology
  • 発表場所: Lake Louse
  • 年月日: 20070728-20070801
• [学会発表] Assessment of the cryoprotectant permeability and cryoprotectant toxicity of aquaporin 3-expressing immature medaka (Oryzias latipes) oocytes (2007)
  • 著者名/発表者名: Delgado M. Valdez Jr., Seki Shinsuke, Takao Hara, Naoya Saida, Yu Nishikado, Kasai Magosaburo, Edashige Keisuke
  • 学会等名: The 44th Annual Meeting of the Society for Cryobiology
  • 発表場所: Lake Louse
  • 年月日: 20070728-20070801
• [学会発表] The role of urea transporters in the movement of cryoprotectants across the plasma membrane in mouse morulae (2007)
  • 著者名/発表者名: Keisuke Edashige, Satoshi Ohta, Tomomi Kajino, Tatsunaga Kuwano, Bo Jin, Delgado M. Valdez Jr., Shinsuke Seki, Magosaburo Kasai
  • 学会等名: The 44th Annual Meeting of the Society for Cryobiology
  • 発表場所: Lake Louse
  • 年月日: 20070728-20070801
• [学会発表] The role of aquaporin-3 in the movement of water and cryoprotectants in mouse morulae (2007)
  • 著者名/発表者名: Keisuke Edashige, Satoshi Ohta, Mitsunobu Tanaka, Tatsunaga Kuwano, Delgado M. Valdez Jr., Takao Hara, Bo. Jin, Sei-ichi Takahashi, Shinsuke Seki, Chihiro Koshimoto, Magosaburo Kasai
  • 学会等名: The 5th International Conference of Aquaporin
  • 発表場所: Nara
  • 年月日: 20070713-20070716
• [学会発表] Intracellular ice formation in vitrified mouse morulae during warming (2007)
  • 著者名/発表者名: Bo Jin, Kenji Kusanagi, Makiko Ueda, Keisuke Edashige, Magosaburo Kasai
  • 学会等名: The 33rd Annual Conference of the International Embryo Transfer Society
  • 発表場所: Kyoto
  • 年月日: 20070107-20070109
• [図書] Movement of water and cryoprotectants in mouse oocytes and embryos at different stages: relevance to cryopreservation (2010)
  • 著者名/発表者名: Magosaburo Kasai, Keisuke Edashige
  • 総ページ数: 16-23
  • 出版者: Cambridge University Press
• [図書] Vitrification in animal reproduction: vitrification of embryos using conventional straws with an ethylene glycol-based solutions (2007)
  • 著者名/発表者名: Magosaburo Kasai, Keisuke Edashige
  • 総ページ数: 75-85
  • 出版者: Informa Healthcare, London