Understanding of molecular mechanisms of chemically-induced toxicities and risk evaluation using zebrafish

• Project/Area Number: 15H05334
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: 放射線・化学物質影響科学
• Research Institution: Obihiro University of Agriculture and Veterinary Medicine
• Principal Investigator: Kubota Akira 帯広畜産大学, 畜産学部, 准教授 (60432811)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥21,450,000 (Direct Cost: ¥16,500,000、Indirect Cost: ¥4,950,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥8,970,000 (Direct Cost: ¥6,900,000、Indirect Cost: ¥2,070,000); Fiscal Year 2015: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
• Keywords: トキシコロジー / ゼブラフィッシュ / 細胞内受容体 / 発生毒性 / 内分泌撹乱 / 環境汚染物質 / エストロゲン受容体 / シトクロムP450 / ビスフェノール類 / in silico解析 / リン酸エステル系難燃剤 / 化学物質 / CYP

Outline of Final Research Achievements

The present study aimed at understanding biological effects of a variety of chemicals and their molecular mechanisms, with particular emphasis on signaling pathways involving cellular receptors, using zebrafish as a model organism. Some of the biological effects identified include cytochrome P450 induction, cardiovascular toxicity, and endocrine disruption, through the cellular receptors such as pregnane X receptor (PXR), aryl hydrocarbon receptor type 2 (AHR2) and estrogen receptors (ERs). The current study also suggest that in silico simulations of interaction between chemicals and ER subtypes can predict in vivo potency of the ER-mediated response by untested chemical substances.

Academic Significance and Societal Importance of the Research Achievements

動物愛護の観点から、化学物質の有害性評価において動物実験代替法の利用を促進する活動が拡大する中で、ゼブラフィッシュを用いた本研究の成果は、動物愛護に寄与し多数の化学物質をハイスループットに評価したものとして学術的・社会的意義は大きい。さらに本研究の成果は、細胞内受容体を介した作用が疑われる医薬品等の安全性評価や環境汚染物質の生態毒性試験法として発展的に活用することも期待され、医療・環境分野における化学物質リスク評価の指針となるだけでなく、わが国の化学物質管理政策にも資すると考えられる。

Research Products

• [Int'l Joint Research] Woods Hole Oceanographic Institution(米国)
• [Int'l Joint Research] University of Antwerp(ベルギー)
• [Int'l Joint Research] Woods Hole Oceanographic Institution(米国)
• [Int'l Joint Research] Chinese Academy of Fishery Sciences(中国)
• [Int'l Joint Research] ウッズホール海洋研究所(米国)
• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Journal Article] Transcriptional profiling of cytochrome P450 genes in the liver of adult zebrafish, <i>Danio rerio</i> (2019)
  • Author(s): Akira Kubota, Yusuke K. Kawai, Natsumi Yamashita, Jae Seung Lee, Daisuke Kondoh, Shuangyi Zhang, Yasunobu Nishi, Kazuyuki Suzuki, Takio Kitazawa, Hiroki Teraoka
  • Journal Title: J. Toxicol. Sci. Volume: 44 Issue: 5 Pages: 347-356
  • DOI: 10.2131/jts.44.347
  • NAID: 130007642915
  • ISSN: 0388-1350
  • Peer Reviewed / Open Access
• [Journal Article] Ecotoxicology of Dioxins in Wild Avian Species (2018)
  • Author(s): 久保田 彰
  • Journal Title: Japanese Journal of Zoo and Wildlife Medicine Volume: 22 Issue: 4 Pages: 63-67
  • DOI: 10.5686/jjzwm.22.63
  • NAID: 130006731441
  • ISSN: 1342-6133
  • Year and Date: 2018-05-04
  • Peer Reviewed
• [Journal Article] The evolution of UDP-glycosyl/glucuronosyltransferase 1E (UGT1E) genes in bird lineages is linked to feeding habits but UGT2 genes is not (2018)
  • Author(s): Yusuke K. Kawai, Yoshinori Ikenaka, Mayumi Ishizuka, Akira Kubota
  • Journal Title: PLOS ONE Volume: 13 Pages: 1-13
  • DOI: 10.1371/journal.pone.0205266
  • NAID: 120006540774
  • Peer Reviewed / Open Access
• [Journal Article] 十勝の台風被害による飼料用トウモロコシサイレージおよび牧草サイレージのかび毒汚染の実態 (2018)
  • Author(s): 川島千帆、久保田彰
  • Journal Title: 北海道畜産草地学会報 Volume: 6 Pages: 29-34
  • NAID: 40021519058
• [Journal Article] Dexamethasone-induced hepatomegaly and steatosis in larval zebrafish (2017)
  • Author(s): Guojun Yin, Liping Cao, Jinliang Du, Rui Jia, Takio Kitazawa, Akira Kubota, Hiroki Teraoka
  • Journal Title: J. Toxicol. Sci. Volume: 42 Issue: 4 Pages: 455-459
  • DOI: 10.2131/jts.42.455
  • NAID: 130005765960
  • ISSN: 0388-1350
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biological effects of 6-formylindolo[3,2-b]carbazole (FICZ) in vivo are enhanced by loss of CYP1A function in an Ahr2-dependent manner (2016)
  • Author(s): Wincent, E., Kubota, A., Timme-Laragy, A. R., J&ouml;nsson, M. E., Hahn, M. E. and Stegeman, J. J.
  • Journal Title: Biochemical Pharmacology Volume: 110-111 Pages: 117-129
  • DOI: 10.1016/j.bcp.2016.04.012
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Protective effect of prostacyclin against pre-cardiac edema caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin and a thromboxane receptor agonist in developing zebrafish. (2016)
  • Author(s): Nijoukubo, D., Tanaka, Y., Okuno, Y., Yin, G., Kitazawa, T., Peterson, R. E., Kubota, A. and Teraoka, H.
  • Journal Title: Chemosphere Volume: 156 Pages: 111-117
  • DOI: 10.1016/j.chemosphere.2016.04.107
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Cytochrome P450 20A1 in zebrafish: Cloning, regulation and potential involvement in hyperactivity disorders (2016)
  • Author(s): Benjamin Lemaire, Akira Kubota, Conor M. O'Meara, David C. Lamb, Robert L. Tanguay, Jared V. Goldstone, John J. Stegeman
  • Journal Title: Toxicology and Applied Pharmacology Volume: 296 Pages: 73-84
  • DOI: 10.1016/j.taap.2016.02.001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Functional characterization of zebrafish cytochrome P450 1 family proteins expressed in yeast (2015)
  • Author(s): Stegeman, J. J., Behrendt, L., Woodin, B. R., Kubota, A., Lemaire, B., Pompon, E., Goldstone, J. V. and Urban, P.
  • Journal Title: Biochimica et Biophysica Acta Volume: 1850(11) Pages: 2340-2352
  • DOI: 10.1016/j.bbagen.2015.07.010
  • Peer Reviewed / Int'l Joint Research
• [Presentation] In vivo measurement and in silico prediction of estrogen-like potency of bisphenol A and its analogues in zebrafish (2019)
  • Author(s): Akira Kubota, Jae Seung Lee, Yumi Wakayama, Michiko Nakamura, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata, Yusuke Kawai
  • Organizer: SOT 58th Annual Meeting and ToxExpo
  • Int'l Joint Research
• [Presentation] In vivo measurement and in silico prediction of estrogenic and anti-estrogenic potency of bisphenol A and its analogues in zebrafish (2019)
  • Author(s): Akira Kubota, Jae Seung Lee, Yumi Wakayama, Michiko Nakamura, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Yusuke Kawai
  • Organizer: International Chemical Hazard Symposium in Hokkaido
  • Int'l Joint Research / Invited
• [Presentation] Assessment of estrogenic and anti-estrogenic potency of bisphenol A replacement in zebrafish (2018)
  • Author(s): Jae Seung Lee, Yumi Wakayama, Michiko Nakamura, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata, Yusuke Kawai, Akira Kubota
  • Organizer: 第27回環境化学討論会
• [Presentation] ゼブラフィッシュを用いた生態毒性試験について (2018)
  • Author(s): 久保田彰
  • Organizer: 第12回生態影響試験実習セミナー
  • Invited
• [Presentation] ゼブラフィッシュを用いたビスフェノールA代替物質の内分泌攪乱作用の評価 (2018)
  • Author(s): 久保田彰、Jae Seung Lee、若山裕己、中村倫子、川合佑典、芳之内結加、岩田久人、平野将司、中田晴彦
  • Organizer: 第45回日本毒性学会学術年会
• [Presentation] 十勝の台風被害による飼料用トウモロコシサイレージのかび毒汚染の実態 (2018)
  • Author(s): 久保田 彰、Jae Seung Lee、川島千帆
  • Organizer: 第45回日本毒性学会学術年会
• [Presentation] Evaluating estrogenic and anti-estrogenic potency of bisphenol A analogues in vivo and in silico using zebrafish (2018)
  • Author(s): Akira Kubota, Yumi Wakayama, Jae Seung Lee, Michiko Nakamura, Yusuke Kawai, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata
  • Organizer: 38th International Symposium on Halogenated Persistent Organic Pollutants
  • Int'l Joint Research
• [Presentation] タンチョウにおける異物代謝関連遺伝子の同定と定量 (2018)
  • Author(s): 川合佑典、伊藤究、坂本梨果子、吉野智生、飯間裕子、松本文雄、久保田彰
  • Organizer: 第161回日本獣医学会学術集会
• [Presentation] In silico prediction and in vivo measurement of estrogenic activity of bisphenol analogues in zebrafish (2018)
  • Author(s): Akira Kubota, Yumi Wakayama, Michiko Nakamura, Yusuke Kawai, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata
  • Organizer: 57th Society of Toxicology Annual Meeting
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュ胚を用いたin silicoおよびin vivo解析によるビスフェノール類のエストロゲン様作用の評価 (2017)
  • Author(s): 川合佑典、中村倫子、若山裕己、森田友理、芳之内結加、平野将司、中田晴彦、岩田久人、久保田 彰
  • Organizer: 第26回環境化学討論会
• [Presentation] In vivo and in silico assessment of estrogen-like effects of bisphenol analogues in zebrafish (2017)
  • Author(s): Yumi Wakayama, Michiko Nakamura, Yuri Morita, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata, Yusuke Kawai, Akira Kubota
  • Organizer: 19th International Symposium on Pollutant Responses in Marine Organisms
  • Int'l Joint Research
• [Presentation] Evolution of avian UGT family genes in relation to feeding habit (2017)
  • Author(s): Yusuke Kawai, Yoshinori Ikenaka, Mayumi Ishizuka, Akira Kubota
  • Organizer: 19th International Symposium on Pollutant Responses in Marine Organisms
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュをモデルとしたin vivoおよびin silico解析によるビスフェノール類のエストロゲン様作用の評価 (2017)
  • Author(s): 中村倫子、若山裕己、芳之内結加、森田友理、岩田久人、川合佑典、久保田 彰
  • Organizer: 第44回日本毒性学会学術年会
• [Presentation] 有機リン系難燃剤およびその代謝物による発達期ゼブラフィッシュに対する影響 (2017)
  • Author(s): 森田友理、若山裕己、芳之内結加、岩田久人、川合佑典、久保田 彰
  • Organizer: 第44回日本毒性学会学術年会
• [Presentation] In silico prediction and in vivo measurement of estrogenic activity of bisphenol analogues in zebrafish (2017)
  • Author(s): Akira Kubota, Yumi Wakayama, Michiko Nakamura, Yusuke Kawai, Yuka Yoshinouchi, Hisato Iwata, Masashi Hirano, Haruhiko Nakata
  • Organizer: 20th International Conference on Cytochrome P450
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュをモデルとしたin silicoおよびin vivo解析によるビスフェノール類のエストロゲン様作用の評価 (2017)
  • Author(s): 久保田 彰、若山裕己、中村倫子、川合佑典、芳之内結加、岩田久人、平野将司、中田晴彦
  • Organizer: 環境ホルモン学会 第20回研究発表会
• [Presentation] Biological effects of a dysfunctional CYP1A/AHR2 feedback loop in developing zebrafish (2016)
  • Author(s): Akira Kubota, Alicia Timme-Laragy, Maria Jonsson, Mark E. Hahn, John J. Stegeman, Emma Wincent
  • Organizer: 環境ホルモン学会 第19回研究発表会
  • Place of Presentation: 文部科学省 研究交流センター（茨城県つくば市）
  • Year and Date: 2016-12-08
• [Presentation] Reciprocal cross-talk between Ahr2 and Pxr signaling in response to PCB126 and pregnenolone in developing zebrafish (2016)
  • Author(s): Akira Kubota, Jared Goldstone, John Stegeman
  • Organizer: The 9th International PCB Workshop
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
  • Year and Date: 2016-10-09
  • Int'l Joint Research
• [Presentation] 発達期ゼブラフィッシュにおけるCYP1A/AHRフィードバック機構攪乱の影響 (2016)
  • Author(s): 久保田彰，Emma Wincent，John Stegeman
  • Organizer: 第22回日本環境毒性学会研究発表会
  • Place of Presentation: 愛媛大学（愛媛県・松山市）
  • Year and Date: 2016-09-06
• [Presentation] Characterization of a Pxr-null zebrafish (2016)
  • Author(s): Matthew C. Salanga, Akira Kubota, Benjamin Lemaire, Rachel Harbeitner, Daniel Gusenleitner, Stefano Monti, John J. Stegeman, Jared V. Goldstone
  • Organizer: 55th Annual Meeting of Society of Toxicology
  • Place of Presentation: ニューオーリンズ（米国）
  • Year and Date: 2016-03-13
  • Int'l Joint Research
• [Presentation] 発達期ゼブラフィッシュをモデルとした化学物質の毒性発現機構の解析 (2016)
  • Author(s): 久保田彰
  • Organizer: 第7回LaMer特別講演会
  • Place of Presentation: 愛媛大学（愛媛県・松山市）
  • Invited
• [Presentation] ゼブラフィッシュにおける薬物代謝酵素の網羅的解析：魚類とヒト健康影響のモデル動物の観点から (2016)
  • Author(s): 久保田彰
  • Organizer: 環境リスク・健康研究センター特別講演会
  • Place of Presentation: 国立環境研究所（茨城県つくば市）
  • Invited
• [Presentation] 魚類におけるPXR・AHRを介したシトクロムP450発現制御機構の特徴 (2015)
  • Author(s): 久保田 彰, Jared V. Goldstone, John J. Stegeman
  • Organizer: 第21回日本環境毒性学会研究発表会
  • Place of Presentation: 東洋大学白山キャンパス（東京都・文京区）
  • Year and Date: 2015-09-02
• [Presentation] 魚類におけるシトクロムP450の発現制御機構と代謝特性 (2015)
  • Author(s): 久保田 彰, John J. Stegeman
  • Organizer: 第42回日本毒性学会学術年会
  • Place of Presentation: 石川県立音楽堂、金沢市アートホテル、ホテル日航金沢（石川県・金沢市）
  • Year and Date: 2015-06-29
• [Presentation] ゼブラフィッシュのリガンド依存性転写因子とシトクロムP450 (2015)
  • Author(s): 久保田 彰, Jared V. Goldstone, Emma Wincent, John J. Stegeman
  • Organizer: 第42回日本毒性学会学術年会
  • Place of Presentation: 石川県立音楽堂、金沢市アートホテル、ホテル日航金沢（石川県・金沢市）
  • Year and Date: 2015-06-29
• [Presentation] Role of pregnane X receptor and aryl hydrocarbon receptor in transcriptional regulation of PXR, CYP2, and CYP3 genes in developing zebrafish (2015)
  • Author(s): Akira Kubota, Jared V. Goldstone, Benjamin Lemaire, John J. Stegeman
  • Organizer: 19th International Conference on Cytochrome P450
  • Place of Presentation: 国立オリンピック記念青少年総合センター（東京都・渋谷区）
  • Year and Date: 2015-06-12
  • Int'l Joint Research
• [Presentation] Differences in responses to putative PXR agonists associated with PXR alleles in zebrafish embryos (2015)
  • Author(s): Akira Kubota, Jared V. Goldstone, Benjamin Lemaire, Rachel Harbeitner, Lauren Salay, John J. Stegeman
  • Organizer: 18th International symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: トロンハイム（ノルウェー）
  • Year and Date: 2015-05-24
  • Int'l Joint Research
• [Presentation] Reciprocal cross-talk between PXR and AHR2 signaling in response to their putative agonists in developing zebrafish (2015)
  • Author(s): Akira Kubota, Jared V. Goldstone, Benjamin Lemaire, Matthew Takata, Bruce R. Woodin, John J. Stegeman
  • Organizer: 18th International symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: トロンハイム（ノルウェー）
  • Year and Date: 2015-05-24
  • Int'l Joint Research
• [Presentation] The PXR “gene battery” in zebrafish (2015)
  • Author(s): Jared V. Goldstone, Rachel Harbeitner, Akira Kubota, Benjamin Lemaire, John J. Stegeman
  • Organizer: 18th International symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: トロンハイム（ノルウェー）
  • Year and Date: 2015-05-24
  • Int'l Joint Research
• [Remarks] 帯広畜産大学獣医学研究部門基礎獣医学分野 毒性学研究室ホームページ
  • URL: http://www.obihiro.ac.jp/~toxicology/index.html
• [Remarks] 帯広畜産大学 獣医学研究部門基礎獣医学分野 毒性学研究室 ホームページ
  • URL: http://www.obihiro.ac.jp/~toxicology/index.html