Analysis of molecular mechanisms for environmental sex determination

• Project/Area Number: 24370029
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Morphology/Structure
• Research Institution: Okazaki Research Facilities, National Institutes of Natural Sciences
• Principal Investigator: IGUCHI Taisen 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 教授 (90128588)
• Co-Investigator(Kenkyū-buntansha): MIYAGAWA Shinichi 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡﨑統合バイオサイエンスセンター, 助教 (30404354) ; OGINO Yukiko 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡﨑統合バイオサイエンスセンター, 助教 (00404343)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2014: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2012: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: 性決定 / 環境 / 爬虫類 / 甲殻類 / 温度 / 環境応答 / ワニ / ミジンコ

Outline of Final Research Achievements

Sex determination mechanisms can broadly be categorized by either a genotypic or environmentally driven mechanism. We investigated environmental sex determination (ESD) mechanism using American alligator and watre flea (daphnia) as a models. We analyzed the thermosensitive cation channel, transient receptor potential (TRP), and its role during alligator temperature-dependent sex determination. Our electrophysiological and pharmacological approaches demonstrate that a thermal TRP is involved in the TSD in alligators and it directly activates male differentiation cascade. We also identified several juvenile hormone and ecdysone system-related genes, which are involved in sex determination pathways in Daphnia. The current results provide useful knowledge for understanding the mechanisms of sex determination system both in vertebrates and invertebrates.

Research Products

• [Journal Article] Estrogen receptor 1 (ESR1; ERα), not ESR2 (ERβ), modulates estrogen-induced sex reversal in the American alligator, a species with temperature-dependent sex determination (2015)
  • Author(s): Kohno, S., Bernhard, M.C., Katsu, Y., Zhu, J., Byan, T.A., Doheny, B.M., Iguchi, T. and Guillette, L.J.Jr.
  • Journal Title: Endocrinology Volume: 印刷中 Issue: 5 Pages: 1887-1899
  • DOI: 10.1210/en.2014-1852
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Methyl farnesoate synthesis is necessary for the environmental sex determination in water flea Daphnia pulex (2015)
  • Author(s): Kenji Toyota, Hitoshi Miyakawa, Chizue Hiruta, Kenjiro Furuta, Yukiko Ogino, Tetsuro Shinoda, Norihisa Tatarazako, Shinichi Miyagawa, Joseph R. Shaw, Taisen Iguchi
  • Journal Title: Journal of Insect Physiology Volume: - Pages: 22-30
  • DOI: 10.1016/j.jinsphys.2015.02.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Ionotropic glutamate receptors mediate inducible defense in the water flea Daphnia pulex (2015)
  • Author(s): Miyakawa, H., Sato, T., Colbourne, J.K. and Iguchi, T.
  • Journal Title: PLoS ONE Volume: 10 Issue: 3 Pages: e0121324-e0121324
  • DOI: 10.1371/journal.pone.0121324
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] NMDA receptor activation upstream of methyl farnesoate signaling for short day-induced male offspring production in the water flea, Daphnia pulex (2015)
  • Author(s): Kenji Toyota, Hitoshi Miyakawa, Katsushi Yamaguchi, Shuji Shigenobu, Yukiko Ogino, Norihisa Tatarazako, Shinichi Miyagawa, Taisen Iguchi
  • Journal Title: BMC Genomics Volume: 16 Issue: 1 Pages: 186-186
  • DOI: 10.1186/s12864-015-1392-9
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Diofenolan induces male offspring production through binding to juvenile hormone receptor in Daphnia magna (2015)
  • Author(s): Ryoko Abe, Kenji Toyota, Hitoshi Miyakawa, Haruna Watanabe, Tomohiro Oka, Shinichi Miyagawa, Hiroyo Nishide, Ikuo Uchiyama, Knut Erik Tollefsen, Taisen Iguchi, Norihisa Tatarazako
  • Journal Title: Aquatic Toxicology Volume: 159 Pages: 44-51
  • DOI: 10.1016/j.aquatox.2014.11.015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Establishment of a short-term in vivo screening method for detecting chemicals having juvenile hormone activity using adult Daphnia magna (2014)
  • Author(s): R. Abe, H. Watanabe, M. Yamamuro, T. Iguchi and N. Tatarazako
  • Journal Title: J. Appl. Toxicol Volume: 印刷中 Issue: 1 Pages: 75-82
  • DOI: 10.1002/jat.2989
  • Peer Reviewed / Open Access
• [Journal Article] Targeted gene disruption by use of transcription activator-like effector nuclease (TALEN) in the water flea Daphnia pulex (2014)
  • Author(s): Chizue Hiruta, Yukiko Ogino, Tetsushi Sakuma, Kenji Toyota, Shinichi Miyagawa, Takashi Yamamoto, Taisen Iguchi
  • Journal Title: BMC Biotechnology Volume: 14 Issue: 1 Pages: 95-95
  • DOI: 10.1186/s12896-014-0095-7
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Roles of ecdysteroids for progression of reproductive cycle in the fresh water crustacean Daphnia magna (2014)
  • Author(s): Eri Sumiya, Yukiko Ogino, Hitoshi Miyakawa, Chizue Hiruta, Kenji Toyota, Shinichi Miyagawa, Taisen Iguchi
  • Journal Title: Frontiers in Zoology Volume: 11 Issue: 1 Pages: 60-60
  • DOI: 10.1186/s12983-014-0060-2
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The reproductive hormone cycle of adult female American alligators from a Barrier island population. Reproduction (2014)
  • Author(s): Hamlin, H.J., Lowers, R.H., Kohno, S., Mitsui-Watanabe, N., Amano, H., Hara, A., Ohta, Y., Miyagawa, S., Iguchi, T. and Guillette, L.J.Jr.
  • Journal Title: Reproduction Volume: 147 Issue: 6 Pages: 855-863
  • DOI: 10.1530/rep-14-0031
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Molecular impact of juvenile hormone agonists on neonatal Daphnia magna. (2014)
  • Author(s): Toyota, K., Kato, Y., Miyakawa, H., Yatsu, R., Mizutani, T., Ogino, Y., Miyagawa, S., Watanabe, H., Nishide, H., Uchiyama, I., Tatarazako, N. and Iguchi, T.
  • Journal Title: Journal of Applied Toxicology Volume: 34 Issue: 5 Pages: 537-544
  • DOI: 10.1002/jat.2922
  • Peer Reviewed
• [Journal Article] Gonadal differentiation in reptiles exhibiting environmental sex determination. (2014)
  • Author(s): Kohno, S., Parrott, B. B., Yatsu, R., Miyagawa, S., Moore, B. C., I guchi, T., and Guillette Jr, L. J.
  • Journal Title: Sexual Development Volume: Vol.0 Issue: 5 Pages: 208-226
  • DOI: 10.1159/000358892
  • Peer Reviewed
• [Journal Article] Development of a microinjection system for RNA interference in the water flea Daphnia pulex (2013)
  • Author(s): Chizue Hiruta, Kenji Toyota, Hitoshi Miyakawa, Yukiko Ogino, Shinichi Miyagawa, Norihisa Tatarazako, Joseph R Shaw, Taisen Iguchi
  • Journal Title: BMC Biotechnology Volume: 13(96) Issue: 1 Pages: 96-96
  • DOI: 10.1186/1472-6750-13-96
  • Peer Reviewed
• [Journal Article] The mutation in methoprene-tolerant alters juvenile hormone use by insects and crustaceans (2013)
  • Author(s): Hitoshi Miyakawa, Kenji Toyota, Ikumi Hirakawa, Yukiko Ogino, Shinichi Miyagawa, Shigeto Oda, Norihisa Tatarazako, Toru Miura, John K. Colbourne, Taisen Iguchi
  • Journal Title: Nature Communications Volume: ncomms2868 Issue: 1 Pages: 1856-1856
  • DOI: 10.1038/ncomms2868
  • Peer Reviewed
• [Journal Article] Molecular cloning of doublesex genes of four cladocera (water flea) species (2013)
  • Author(s): Kenji Toyota, Yasuhiko Kato, Masaru Sato, Naomi Sugiura, Shinichi Miyagawa, Hitoshi Miyakawa, Hajime Watanabe, Shigeto Oda, Yukiko Ogino, Chizue Hiruta, Takeshi Mizutani, Norihisa Tatarazako, Susanne Paland, Craig Jackson, John K. Colbourne, Taisen Iguchi
  • Journal Title: BMC Genomics Volume: 14:239 Issue: 1 Pages: 239-239
  • DOI: 10.1186/1471-2164-14-239
  • Peer Reviewed
• [Journal Article] Molecular cloning and characterization of the corticoid receptors from the American alligator. (2013)
  • Author(s): Oka K, Kohno S, Urushitani H, Guillette LJ, Ohta Y, Iguchi T, Katsu Y
  • Journal Title: Mol. Cell. Endocrinol. Volume: 365 Issue: 2 Pages: 153-161
  • DOI: 10.1016/j.mce.2012.10.014
  • NAID: 120005228364
  • Peer Reviewed
• [Journal Article] Sequencing three crocodilian genomes to illuminate the evolution of archosaurs and amniotes (2012)
  • Author(s): St John JA, et al.
  • Journal Title: Genome Biol. Volume: 13 Pages: 415-415
  • Peer Reviewed
• [Journal Article] Technique for the collection of clear urine from the Nile crocodile (Crocodylus niloticus) (2012)
  • Author(s): Myburgh JG, et al.
  • Journal Title: J S Afr Vet Assoc Volume: 83
  • Peer Reviewed
• [Presentation] Analusis of juvenile hormone as a sex determining factor in water flea (2014)
  • Author(s): Miyagawa, S., Toyota, K. and Iguchi, T.
  • Organizer: 第17回環境ホルモン学会
  • Place of Presentation: 東京大学（東京都文京区）
  • Year and Date: 2014-12-09 – 2014-12-10
• [Presentation] 若ホルモン短期検出法を用いた農薬の昆虫ホルモン作用の検出 (2014)
  • Author(s): 阿部良子、高信瞳、渡部春奈、井口泰泉、鑪迫典久
  • Organizer: 第17回環境ホルモン学会
  • Place of Presentation: 東京大学（東京都文京区）
  • Year and Date: 2014-12-09 – 2014-12-10
• [Presentation] Analyses of ecdysteroids during reproductive cycle and embryogenesis in Daphnia magna. (2014)
  • Author(s): Sumiya, E., Ogino, Y., Miyakawa, H., Hiruta, C., Toyota, K., Miyakawa, S. and Iguchi, T.
  • Organizer: 第39回日本比較内分泌学会第8回ISAREN合同大会
  • Place of Presentation: 岡崎カンファレンスセンター（愛知県岡崎市）
  • Year and Date: 2014-11-07 – 2014-11-09
• [Presentation] Temperature-dependent sex determination and characterization of androgen system in American alligator. (2014)
  • Author(s): Miyagawa, S., Yatsu, R., Kohno, S., Guillette, LJr., Iguchi, T.
  • Organizer: 第39回日本比較内分泌学会第8回ISAREN合同大会
  • Place of Presentation: 岡崎カンファレンスセンター（愛知県岡崎市）
  • Year and Date: 2014-11-07 – 2014-11-09
• [Presentation] Environmental sex determination in the water flea (2014)
  • Author(s): Iguchi, T.
  • Organizer: SEB Annual Meeting
  • Place of Presentation: マンチェスター大学（マンチェスター、イギリス）
  • Year and Date: 2014-07-01 – 2014-07-04
  • Invited
• [Presentation] Establishment of a short-term, in vivo screening method for detecting chemicals with juvenile hormone activity using adult Daphnia magna. (2014)
  • Author(s): Abe, R., Watanabe, H., Iguchi, T., Tatarazako, N.
  • Organizer: 10th International Conference on Juvenile Hormones (JHX)
  • Place of Presentation: 研究交流センター（茨城県つくば市）
  • Year and Date: 2014-06-09 – 2014-06-13
• [Presentation] Analysis of the reception and molecular signaling downstream of a crustacean juvenile hormone, methyl farnesoate, in Daphnia pulex (2014)
  • Author(s): Miyakawa, H., Sato, M., Ogino, Y.,Miyagawa, S., Colbourne, J.K., Iguchi, T.
  • Organizer: 10th International Conference on Juvenile Hormones (JHX)
  • Place of Presentation: 研究交流センター（茨城県つくば市）
  • Year and Date: 2014-06-09 – 2014-06-13
• [Presentation] Approach to establishing a tool for targeted gene disruption by using TALEN in the water flea Daphnia pulex (2014)
  • Author(s): Hiruta, C., Ogino,Y., Toyota, K., Iguchi, T.
  • Organizer: 第47回環境ホルモン学会
  • Place of Presentation: ウインクあいち（愛知県名古屋市）
  • Year and Date: 2014-05-27 – 2014-05-30
• [Presentation] A Single amino acid substitution in the Methoprene-tolerant (Met) alters juvenile hormone use by insects and crustaceans (2014)
  • Author(s): H. Miyakawa, K. Toyota, I. Hirakawa, Y. Ogino, S. Miyagawa, S. Oda, N. Tatarazako, T. Miura, J.K. Colbourne, T. Iguchi
  • Organizer: EMBO Conference; Daphnia Genomics Consortium 2014
  • Place of Presentation: バーミンガム大学（バーミンガム、イギリス）
• [Presentation] Analysis of juvenile hormone as a sex-determining hormone in Daphnia pulex (2014)
  • Author(s): K. Toyota, H. Miyakawa, C. Hiruta, E. Sumiya, Y. Ogino, S. Miyagawa, T. Iguchi
  • Organizer: EMBO Conference; Daphnia Genomics Consortium 2014
  • Place of Presentation: バーミンガム大学（バーミンガム、イギリス）
• [Presentation] Cloning and their expression pattern of doublesex genes of cladocera (2014)
  • Author(s): S. Miyagawa, K. Toyota, H. Miyakawa, C. Hiruta, Y. Kato, H. Watanabe, N. Tatarazako, S. Paland, C. Jackson, J.K. Colbourne, T. Iguchi
  • Organizer: EMBO Conference; Daphnia Genomics Consortium 2014
  • Place of Presentation: バーミンガム大学（バーミンガム、イギリス）
• [Presentation] pproach to establishing tools for gene functional analysis and cryopreservation in Daphnia pulex (2014)
  • Author(s): C. Hiruta, K. Toyota, H. Miyakawa, S. Miyagawa, Y. Ogino, N. Tatarazako, J.R. Shaw, D. Tanaka, T. Iguchi
  • Organizer: EMBO Conference; Daphnia Genomics Consortium 2014
  • Place of Presentation: バーミンガム大学（バーミンガム、イギリス）
• [Presentation] cdysteroids are involved in regulatory mechanismof a reproductive cycle in D. Magna (2014)
  • Author(s): E. Sumiya, Y. Ogino, H. Miyakawa, C. Hiruta, K. Toyota, S. Miyagawa, T. Iguchi
  • Organizer: EMBO Conference; Daphnia Genomics Consortium 2014
  • Place of Presentation: バーミンガム大学（バーミンガム、イギリス）
• [Presentation] ミジンコにおける内分泌かく乱作用メカニズムの解析 (2013)
  • Author(s): 井口泰泉
  • Organizer: 環境ホルモン学会第16回研究発表会
  • Place of Presentation: 東京大学（東京都、文京区）
  • Invited
• [Presentation] cdysteroid in higher titer inhibits molting and reproduction cycle in Daphnia magna. (2013)
  • Author(s): E. Sumiya, Y. Ogino, H. Miyakawa, C. Hiruta, K. Toyota, S. Miyagawa, T. Iguchi
  • Organizer: 環境ホルモン学会第16回研究発表会
  • Place of Presentation: 東京大学（東京都、文京区）
• [Presentation] ミジンコDaphnia pulexにおける6種類の幼若ホルモン様物質の暴露による幼若ホルモン応答遺伝子の網羅的探索 (2013)
  • Author(s): 宮川一志、井口泰泉
  • Organizer: 環境ホルモン学会第16回研究発表会
  • Place of Presentation: 東京大学（東京都、文京区）
• [Presentation] Cloning of alligator androgen receptor and its response to hormones and chemicals (2013)
  • Author(s): S. Miyagawa, S. Kohno, R. Yatsu, Y. Ogino, Y. Ohta, LJ Guillette .Jr., T. Iguchi
  • Organizer: 環境ホルモン学会第16回研究発表会
  • Place of Presentation: 東京大学（東京都、文京区）
• [Presentation] ミジンコの性決定ホルモンとしての幼若ホルモンの新規機能の解明 (2013)
  • Author(s): 豊田賢治, 宮川一志, 蛭田千鶴江, 角谷絵里, 荻野由紀子, 宮川信一, 井口泰泉
  • Organizer: 第38回日本比較内分泌学会大会
  • Place of Presentation: 宮崎市民プラザ（宮崎県、宮崎市）
• [Presentation] ワニのアンドロゲン受容体の機能解析 (2013)
  • Author(s): 宮川信一、河野郷通、谷津遼平、勝義直、Louis Guillette Jr.、井口泰泉
  • Organizer: 第38回日本比較内分泌学会大会
  • Place of Presentation: 宮崎市民プラザ（宮崎県、宮崎市）
• [Presentation] 有鱗目におけるエストロゲン受容体の機能解析、および進化的傾向 (2013)
  • Author(s): 谷津遼平、宮川信一、勝義直、井口泰泉
  • Organizer: 第84回日本動物学会
  • Place of Presentation: 岡山大学（岡山県、岡山市）
• [Presentation] ワニの抗ミュラー管ホルモン遺伝子の糖質コルチコイド受容体による発現制御 (2013)
  • Author(s): 岡香織、佐々木麻衣、大田泰彦、井口泰泉、勝義直
  • Organizer: 第84回日本動物学会
  • Place of Presentation: 岡山大学（岡山県、岡山市）
• [Presentation] Development of an RNA interference method using microinjection in the water flea Daphnia pulex, (2013)
  • Author(s): C. Hiruta, K. Toyota, H. Miyakawa, T. Iguchi
  • Organizer: 日本発生生物学会第46回大会
  • Place of Presentation: くにびきメッセ（島根県、松江市）
• [Presentation] ミジンコ類のMetおよびSRCを用いたin vitroにおける幼若ホルモン作用評価系の開発 (2012)
  • Author(s): 宮川一志, ら
  • Organizer: 環境ホルモン学会第15回研究発表会
  • Place of Presentation: 東京大学山上会館(東京都文京区)
  • Year and Date: 2012-12-18
• [Presentation] Analysis of sex determining genes involved in male induction in Cladocerans treated by juvenile hormone agonist (2012)
  • Author(s): Toyota, K., et al.
  • Organizer: 環境ホルモン学会第15回研究発表会
  • Place of Presentation: 東京大学山上会館(東京都文京区)
  • Year and Date: 2012-12-18
• [Presentation] オオミジンコ脱皮ホルモン合成経路遺伝子の脱皮周期における時空間的発現解析 (2012)
  • Author(s): 角谷絵里, ら
  • Organizer: 環境ホルモン学会第15回研究発表会
  • Place of Presentation: 東京大学山上会館(東京都文京区)
  • Year and Date: 2012-12-18
• [Presentation] 節足動物の内分泌かく乱物質に対する短期検出法の開発 (2012)
  • Author(s): 阿部良子, ら
  • Organizer: 環境ホルモン学会第15回研究発表会
  • Place of Presentation: 東京大学山上会館(東京都文京区)
  • Year and Date: 2012-12-18
• [Presentation] オオミジンコの正中線におけるDmagSIM発現細胞の神経細胞タイプの同定 (2012)
  • Author(s): 森田慎一, ら
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場(福岡県福岡市)
  • Year and Date: 2012-12-11
• [Presentation] オオミジンコ付属肢形成におけるNotch Signaling Pathway遺伝子群の機能解析 (2012)
  • Author(s): 上原政樹, ら
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場(福岡県福岡市)
  • Year and Date: 2012-12-11
• [Presentation] オオミジンコDaphnia magnaにおけるHox遺伝子Sex Combs ReducedのRNAi法による機能解析 (2012)
  • Author(s): 大見川沙織, ら
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場(福岡県福岡市)
  • Year and Date: 2012-12-11
• [Presentation] ミジンコ類におけるdouble sex遺伝子の単離及び発現解析 (2012)
  • Author(s): 豊田賢治, ら
  • Organizer: 平成24年度日本動物学会中部支部大会
  • Place of Presentation: 松本市浅間温泉みやま荘(長野県松本市)
  • Year and Date: 2012-11-17
• [Presentation] オオミジンコ脱皮ホルモン合成経路遺伝子の分子系統解析と脱皮周期中の発現変動 (2012)
  • Author(s): 角谷絵里, ら
  • Organizer: 平成24年度日本動物学会中部支部大会
  • Place of Presentation: 松本市浅間温泉みやま荘(長野県松本市)
  • Year and Date: 2012-11-17
• [Presentation] ミジンコ(Daphnia pulex)初期胚へのマイクロインジェクション法の確立に向けて (2012)
  • Author(s): 蛭田千鶴江, ら
  • Organizer: 平成24年度日本動物学会中部支部大会
  • Place of Presentation: 松本市浅間温泉みやま荘(長野県松本市)
  • Year and Date: 2012-11-17
• [Presentation] 節足動物類における幼若ホルモンの受容機構とその進化の解明 (2012)
  • Author(s): 宮川一志, ら
  • Organizer: 平成24年度日本動物学会中部支部大会
  • Place of Presentation: 松本市浅間温泉みやま荘(長野県松本市)
  • Year and Date: 2012-11-17
• [Presentation] Establishment of transactivation qssays using hormone receptors from various animal species for screening of environmental chemicals. (2012)
  • Author(s): Iguchi, T., et al.
  • Organizer: SETAC Asia Pacific
  • Place of Presentation: ホテル日航(熊本県熊本市)(招待講演)
  • Year and Date: 2012-09-24
• [Presentation] オオミジンコ脱皮ホルモン合成経路遺伝子の発現変動 (2012)
  • Author(s): 角谷絵里, ら
  • Organizer: 第83回日本動物学会
  • Place of Presentation: 大阪大学(大阪府豊中市)
  • Year and Date: 2012-09-13
• [Remarks] Iguchi lab.
  • URL: http://www.nibb.ac.jp/bioenv1/index-j.html
• [Remarks] Iguchi Lab.
  • URL: http://www.nibb.ac.jp/bioenv1/index-j.html
• [Remarks] Iguchi Lab on Web