A spatiotemporal SRY action in mammalian gonadal sex determination

• Project/Area Number: 16086203
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: KANAI Yoshiakira The University of Tokyo, 大学院・農学生命科学研究科, 准教授 (30260326)
• Co-Investigator(Kenkyū-buntansha): TSUNEKAWA NAOKI 東京大学, 大学院・農学生命科学研究科, 助教 (50431838)
• Project Period (FY): 2004 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥45,500,000 (Direct Cost: ¥45,500,000); Fiscal Year 2008: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2007: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2006: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2005: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2004: ¥9,100,000 (Direct Cost: ¥9,100,000)
• Keywords: SRY / トランスジェニック / 性決定 / マウス / Sry / FGF9 / WNT4 / sFRP2 / マイクロアレイ / 性転換 / トランスジェニックマウス / 性分化 / 分子カスケード / 生殖腺

Research Abstract

本研究課題により、新たにSry機能解析用のモデル動物の樹立を目的とし、マウスHsp70プロモーターにSryを連結した発現ベクターを用いて、Sry発現様式を改変したトランスジェニック(Tg)マウスの樹立を行った。その結果、ステージ・領域非特異的かつ恒常的にSry発現するXX精巣の性転換ライン(#40、#46)2系統を樹立した。さらに、通常飼育下で繁殖能があり(XX卵巣)、人為的に熱ショック(HS)を与えることにより、卵巣からXX精巣へと性転換させることができるSry誘導モデルマウス(#44)の樹立に初めて成功した。本モデルマウスを用いて、1)性分化初期(11.0~11.25dpc)でのSry発現が精巣誘導に必須であること(精巣誘導の臨界期の決定)、2)初期のSry発現がWnt4の発現の抑制に必須であること、3)SRYのみならずFGF9因子が時空間的なSox9発現制御に重要であること、4)Sry直下のカスケードに雄特異的に性腺にエネルギー供給する分子メカニズムの存在、5)Sry単独では完全な精子形成能を持つセルトリ細胞に分化できないこと(XX/Sryセルトリ細胞の機能不全)、6)Sry誘導により6時間以内に発現誘導されるSRY標的遺伝子を新たに78個の遺伝子を同定-などを見い出した。

Research Products

• [Journal Article] A critical time windows of Sry action in gonadal sex differentiation in mice (2009)
  • Author(s): Hiramatsu, R., Matoba, S., Kanai-Azuma, M., Tsunekawa, N., Katoh-Fukui, Y., Kurohmaru, M., Morohashi, K., Wilhelm, D., Koopman, P., Kanai, Y
  • Journal Title: Development 36(1) Pages: 129-138
• [Journal Article] The Cerebellin 4 Precursor Gene Is a Direct Target of SRY and SOX9 in Mice (2009)
  • Author(s): Bradford, S.T., Hiramatsu, R., Maddugoda, M.P., Bernard, P., Chaboissier, M.C., Sinclair, A., Schedl, A., Harley, V., Kanai, Y., Koopman, P., Wilhelm, D
  • Journal Title: Biol. Reprod 80(6) Pages: 1178-1188
• [Journal Article] Evidence for crucial role of hindgut expansion in directing proper migration of primordial germ cells in mouse early embryogenesis (2009)
  • Author(s): Hara K, Kanai-Azuma M, Uemura M, Shitara H, Taya C, Yonekawa H, Kawakami H, Tsunekawa N, Kurohmaru M, Kanai Y
  • Journal Title: Dev Biol 330(2) Pages: 427-439
  • Peer Reviewed
• [Journal Article] A critical time windows of Sry Action in gonadal sex differentiation in mice. (2009)
  • Author(s): Hiramatsu, et al.
  • Journal Title: Development 136 Pages: 129-138
  • Peer Reviewed
• [Journal Article] The Cerebellin 4 Precursor Gene Is a Direct Target of SRY and SOX9 in Mice. (2009)
  • Author(s): Bradford, et al.
  • Journal Title: Biol Reprod 80 Pages: 1178-1188
  • Peer Reviewed
• [Journal Article] Five azacytidine, a DNA methyltranseferase inhibitor, specifically inhibits the testicualar cord formation and Sertoli cell differentiation, in vitro (2008)
  • Author(s): Mizukami, T., Kanai, Y., Fujisawa, M., Kanai-Azuma, M., Kurohmaru, M., Hayashi, Y
  • Journal Title: Mol. Reprod. Dev 75(6) Pages: 1002-1010
• [Journal Article] Akt signaling promotes derivation of embryonic germ cells from primordial germ cells (2008)
  • Author(s): Kimura, T., Tomooka, M., Yamano, N., Murayama, K., Matoba, S., Umehara, H., Kanai, Y., Nakano, T
  • Journal Title: Development 135(5) Pages: 869-879
  • NAID: 120004837399
• [Journal Article] Establishment of testis-specific SOX9 activation requires high-glucose metabolism in mouse sex differentiation (2008)
  • Author(s): Matoba, S., Hiramatsu, R., Kanai-Azuma, M., Tsunekawa, N., Harikae, K., Kawakami, H., Kurohmaru, M., Kanai, Y
  • Journal Title: Dev Biol 324(1) Pages: 76-87
• [Journal Article] Establishment of testis-specific SOX9 activation requires high-glucose metabolism in mouse sex differentiation. (2008)
  • Author(s): Matoba, et al.
  • Journal Title: Dev Biol 324 Pages: 76-87
  • Peer Reviewed
• [Journal Article] Five azacytidine, a DNA methyltranseferase inhibitor, specifically inhibits the testicualar cord formation and Sertoli cell differentiation, in vitro. (2008)
  • Author(s): Mizukami, T., et. al.
  • Journal Title: Mol. Reprod. Dev. 75 Pages: 1002-1010
  • Peer Reviewed
• [Journal Article] Potency of testicular somatic environment to support spermatogenesis in XX/Sry transgenic male mice (2007)
  • Author(s): Ishii, M., Tachiwana, T., Hoshino, A., Tsunekawa, N., Hiramatsu, R., Matoba, S., Kanai-Azuma, M., Kawakami H., Kurohmaru, M., Kanai, Y
  • Journal Title: Development 134 Pages: 449-454
• [Journal Article] SOX9 regulates prostaglandin D synthase gene transcription in vivo to ensure testis development (2007)
  • Author(s): Wilhelm, D., Hiramatsu, R., Mizusaki, H., Widjaja, L., Combes, A.N., Kanai, Y., Koopman, P
  • Journal Title: J. Biol. Chem 282 Pages: 10553-10560
• [Journal Article] Redundant roles of Sox17 and Sox18 in early cardiovascular development of mouse embryos. Biochem. Biophys (2007)
  • Author(s): Sakamoto, Y., Hara, K., Kanai-Azuma, M., Matsui, T., Miura, Y., Tsunekawa, N., Kurohmaru, M., Saijoh, Y., Koopman, P., Kanai, Y
  • Journal Title: Res. Commun 360(3) Pages: 539-544
• [Journal Article] Isolation, characterization, and in vitro and in vivo differentiation of putative thecal stem cells. Proc. Natl (2007)
  • Author(s): Honda, A., Hirose, M., Hara, K., Matoba, S., Inoue, K., Hiura, H., Miki, H., Kanai, Y., Kono, T., Shinohara, T., Ogura A
  • Journal Title: Acad. Sci. USA 104(30) Pages: 12389-12394
• [Journal Article] Sox17 plays a substantial role in late-stage differentiation of the extraembryonic endoderm in vitro (2007)
  • Author(s): Shimoda, M, Kanai-Azuma, M., Hara, K., Miyazaki, S., Kanai, Y., Monden, M., Miyazaki, J
  • Journal Title: J. Cell Sci 120(21) Pages: 3859-3869
• [Journal Article] Potency of testicular somatic environment to support spermatogenesis in XX/Sry transgenic male mice. (2007)
  • Author(s): Ishii, M., et. al.
  • Journal Title: Development 134 Pages: 449-454
  • Peer Reviewed
• [Journal Article] SOX9 regulates prostaglandin D synthase gene transcription in vivo toensure testis development. (2007)
  • Author(s): Wilhelm, D., et. al.
  • Journal Title: J. Biol. Chem. 282 Pages: 10553-10560
  • Peer Reviewed
• [Journal Article] SOX9 regulates prostaglandin D synthase gene transcription in vivo to ensure testis development. (2007)
  • Author(s): Wilhelm, D。 et al.
  • Journal Title: J. Biol. Chem. (印刷中)
• [Journal Article] Potency of testicular somatic environment to support spermatogenesis in XX/Sry transgenic male mice. (2007)
  • Author(s): Ishii, M. et al.
  • Journal Title: Development 134 Pages: 449-454
• [Journal Article] Crucial transcription factors in endoderm and embryonic gut development are expressed in gut-like structures from mouse ES cells (2006)
  • Author(s): Matsuura, R., Kogo, H., Ogaeri, T., Miwa, T., Kuwahara, M., Kanai, Y., Nakagawa, T., Kuroiwa, A., Fujimoto, T., Torihashi, S
  • Journal Title: Stem Cells 24(3) Pages: 624-630
  • NAID: 120000974103
• [Journal Article] Redundant roles of Sox17 and Sox18 in postnatal angiogenesis in mice (2006)
  • Author(s): Matsui, M., Kanai-Azuma, M., Hara K., Matoba, S., Hiramatsu, R., Kawakami, H., Kurohmaru, M., Koopman, P., Kanai Y
  • Journal Title: J. Cell Sci 119(17) Pages: 3513-3526
• [Journal Article] Crucial role of the small GTPase ARF6 in hepatic cord formation during liver development (2006)
  • Author(s): Suzuki, T., Kanai Y., Hara, T., Sasaki, J., Sasaki, T., Kohara, M., Maehama, T., Taya, C., Shitara, H., Yonekawa, H., Frohman, M.A.., Yokozeki, T., Kanaho, Y
  • Journal Title: Mol. Cell. Biol 26(16) Pages: 6149-6156
• [Journal Article] Identification of Sox17 as a transcription factor that regulates oligodendrocyte development (2006)
  • Author(s): Sohn, J., Natale, J., Chew, L-J., Belachew, S., Cheng, Y., Aguirre, A., Lytle, J., Kanai-Azuma, M., Kanai, Y., Gallo, V
  • Journal Title: J. Neurosci 26(38) Pages: 9722-9735
• [Journal Article] Expression Pattern of alphavbeta3 and alphavbeta5 Integrin mRNA in Mouse Fetal Gonads. (2006)
  • Author(s): Ishii, M. et al.
  • Journal Title: J Reprod Dev. 52 Pages: 461-468
• [Journal Article] 哺乳類の性決定遺伝子Sry.「特集 : 雄と雌の生物学」 (2006)
  • Author(s): 的場, 平松, 金井
  • Journal Title: 細胞工学 Pages: 369-373
• [Journal Article] 哺乳類の性決定遺伝子Sry (2006)
  • Author(s): 的場, 平松, 金井
  • Journal Title: 細胞工学 25・4(印刷中)
• [Journal Article] Influence on spatiotemporal patterns of a male-specific Sox9 activation by ectopic Sry Expression during early phases of testis differentiation in mice (2005)
  • Author(s): Kidokoro, T., Matoba S., Hiramatsu, R., Fujisawa, M., Kanai-Azuma, M., Taya, C., Kurohmaru, M., Kawakami, H., Hayashi, Y., Kanai, Y., Yonekawa, H
  • Journal Title: Dev. Biol 278(2) Pages: 511-525
• [Journal Article] A novel Sry-downstream cellular event which preserves the readily available energy source of glycogen in mouse sex differentiation (2005)
  • Author(s): Matoba, S., Kanai, Y., Kidokoro, T., Kanai-Azuma, M., Kawakami, H., Hayashi, Y., Kurohmaru, M
  • Journal Title: J. Cell Sci 118(7) Pages: 1449-1459
• [Journal Article] From SRY to SOX9: mammalian testis differentiation (2005)
  • Author(s): Kanai, Y., Hiramatsu, R., Matoba, S., Kidokoro, T
  • Journal Title: J. Biochem 138 Pages: 13-19
  • NAID: 10017345917
• [Journal Article] Adhesion activity of fetal gonadal cells to EGF and discoidin domains of MFG-E8, a secreted integrin-binding protein which is transiently expressed in mouse early gonadogenesis (2005)
  • Author(s): Ishii, M., Kanai, Y., Kanai-Azuma, M., Tajima, Y., Tay, T.W., Kidokoro, T., Sanai, Y., Kurohmaru, M., Hayashi, Y
  • Journal Title: Anat. Embryol 209(6) Pages: 485-494
• [Journal Article] A novel Sry-downstream cellular event which preserves the readily available energy source of glycogen in mouse sex differentiation. (2005)
  • Author(s): Matoba, S. et al.
  • Journal Title: J.Cell Sci. 118・7 Pages: 1449-1459
• [Journal Article] Influence on spatiotemporal patterns of a male-specific Sox9 activation by ectopic Sry expression during early phases of testis differentiation in mice (2005)
  • Author(s): Kidokoro, T. et al.
  • Journal Title: Dev.Biol. 278・2 Pages: 511-525
• [Journal Article] From SRY to SOX9 : mammalian testis differentiation (2005)
  • Author(s): Kanai, Y. et al.
  • Journal Title: J.Biochem. 138・1 Pages: 13-19
  • NAID: 10017345917
• [Journal Article] Adhesion activity of fetal gonadal cells to EGF and discoidin domains of MFG-E8, a secreted integrin-binding protein which is transiently expressed in mouse early gonadogenesis. (2005)
  • Author(s): Ishii, M. et al.
  • Journal Title: Anat Embryol 209・6 Pages: 485-494
• [Journal Article] Influence on spatiotemporal patterns of a male-specific Sox9 activation by ectopic Sry expression during early phases of testis differentiation in mice. (2005)
  • Author(s): Kidokoro, T. et al.
  • Journal Title: Dev.Biol. 278 Pages: 511-525
• [Presentation] A critical time window of Sry action in gonadal sex determiantion (2008)
  • Author(s): Kanai, Y
  • Organizer: The 41st Annual Meeting for the Japanese Society of Developmental Biologists (Jointly sponsored by the International Society of Developmental Biologists)
  • Place of Presentation: Tokushima, Japan
• [Presentation] 哺乳類の性決定および性分化機構について. シンポジウム「脊椎動物の性決定・性分化とステロイドホルモン」 (2008)
  • Author(s): 金井
  • Organizer: 第79回日本動物学会
  • Place of Presentation: 福岡大学(福岡)
• [Presentation] A critical time window of Sry in gonadal sex determination in mice (2008)
  • Author(s): Kanai Y
  • Organizer: The 2nd SOX Meeting (organized by Hisato Kondoh and Michael Wegner)
  • Place of Presentation: Awaji, Japan
• [Presentation] Haploinsufficiency of Sox17 Results in Defective Maturation of Fetal Hepatocytes in C57BL6 Mice (2008)
  • Author(s): Kanai-Azuma M, Kanai Y
  • Organizer: The 2nd SOX Meeting (organized by Hisato Kondoh and Michael Wegner)
  • Place of Presentation: Awaji, Japan
• [Presentation] 性決定遺伝子SRYとその時間/空間的な制御 (2007)
  • Author(s): 金井
  • Organizer: 東海実験動物研究会
  • Place of Presentation: エーザイ川島工園(岐阜)
• [Presentation] Sryによる性の人為的な制御と精子形成能.シンポジウム「精巣を用いた発生工学の最前線」 (2007)
  • Author(s): 金井克晃
  • Organizer: 第143回日本獣医学会.
  • Place of Presentation: つくば国際会議場.
• [Presentation] 哺乳類の性決定遺伝子SRY. シンポジウム「性分化と生殖細胞の増殖, 分化」 (2006)
  • Author(s): 金井
  • Organizer: 第111回日本解剖学会総会
  • Place of Presentation: 北里大学(神奈川)
• [Presentation] Potency of XX/ Sry Sertoli cells to support spermatogenesis in sex-reversed XX male mice (2006)
  • Author(s): Kanai Y
  • Organizer: The 4th International Symposium on Vertebrate Sex Determination (organized by Val Lance, Mark Bogart, Blanche Capel and Richard Behringer)
  • Place of Presentation: Hawaii, USA
• [Presentation] XX/Sry Sertoli cells are not capable of supporting spermatogenesis in mice (2005)
  • Author(s): Kanai Y
  • Organizer: The 1st SOX Meeting (organized by Hisato Kondoh and Peter Koopman)
  • Place of Presentation: Cairns, Australia
• [Presentation] Redundant roles of Sox17 and Sox18 in postnatal angiogenesis in mice (2005)
  • Author(s): Kanai-Azuma M, Kanai Y
  • Organizer: The 1st SOX Meeting (organized by Hisato Kondoh and Peter Koopman)
  • Place of Presentation: Cairns, Australia
• [Presentation] 哺乳類の性決定遺伝子SRYの機能. シンポジウム「受精成立研究の最前線～性決定からエピジェネティクスまで～」 (2005)
  • Author(s): 金井
  • Organizer: 第76回日本動物学会
  • Place of Presentation: つくば国際会議場(茨城県筑波市)
• [Presentation] Sryによる性の人為的な制御と精子形成能. シンポジウム「精巣を用いた発生工学の最前線」
  • Author(s): 金井
  • Organizer: 第143回日本獣医学会
  • Place of Presentation: つくば国際会議場(茨城)
• [Presentation] 講演会哺乳類の遺伝的な一次性決定とその後の可塑性
  • Author(s): 金井
  • Place of Presentation: 早稲田先端医科学センター(東京)
• [Presentation] Manipulation of the Timing of SRY Activity in Transgenic Mice Defines a Critical Window for Male Sex Determination
  • Author(s): Kanai, Y
  • Organizer: In International symposium for Gonad and Brain Sex Differentiation
  • Place of Presentation: Fukuoka, Japan
• [Book] ホールマウントin situハイブリダイゼーション.「組織細胞化学2008」 (2008)
  • Author(s): 金井, 川上, 原, 金井
  • Publisher: 中西印刷
• [Book] 哺乳類の性決定遺伝子 sry. 「特集:雄と雌の生物学」 (2006)
  • Author(s): 的場, 平松, 金井
  • Publisher: 細胞工学
• [Book] ホールマウント in situハイブリダイゼーション -up to date.分担「組織細胞化学2006」 (2006)
  • Author(s): 金井, 川上, 平松, 金井
  • Publisher: 中西印刷
• [Book] ホールマウントin situハイブリダイゼーション-up to date. 分担「組織細胞化学2006」 (2006)
  • Author(s): 金井, 川上, 平松, 金井
  • Publisher: 中西印刷
• [Book] モデル動物各論.性決定.「モデル動物の作製と維持」 (2004)
  • Author(s): 金井克晃 分担
  • Publisher: エル・アイ・シー
• [Remarks]
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