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魚類の核内受容体PXRシグナル経路の撹乱に関わる新規分子機構とその毒性学的意義

Research Project

Project/Area Number 26881001
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Risk sciences of radiation and chemicals
Research InstitutionObihiro University of Agriculture and Veterinary Medicine

Principal Investigator

久保田 彰  帯広畜産大学, 畜産学部, 准教授 (60432811)

Project Period (FY) 2014-08-29 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsゼブラフィッシュ / PXR / AHR / シトクロムP450 (CYP) / 細胞内受容体 / クロストーク / シグナル伝達 / PCB
Outline of Annual Research Achievements

本年度の成果は、以下のように要約できる。
(1)発達期ゼブラフィッシュにおいてpregnane X receptor(PXR)の標的遺伝子の発現量を変動させる化学物質を探索した。受精後48時間の胚にビスフェノール類(BPA、BPF、BPAF;各0.001μM~10μM)を曝露したところ、いずれもPXR発現量に影響を及ぼさなかったものの、シトクロムP450(CYP)3A65発現量を低下させる傾向を示した。
(2)複数の雌雄成魚のランダム交配から得た受精卵(20個/ペトリ皿)を用いてクロトリマゾール(CLO;3μM)の曝露試験を繰り返し実施したところ、CYP3A65の誘導性に実験間で差異が認められた。CLO応答性およびCLO低応答性を示した実験について、それぞれPXRの塩基配列を解析したところ、一塩基変異(SNPs)の出現パターンに差異が認められた。一例として、184番目のアミノ酸はCLO応答性ではセリン(pxr*1型)、CLO低応答性ではイソロイシン(pxr*2型)が顕在化していた。これらPXRアレルとCLO応答性/低応答性の関連性については今後の課題となった。
(3)ゼブラフィッシュ胚でPXR・aryl hydrocarbon receptor type 2(AHR2)・CYP1/2/3分子種の発現調節に対するPXR・AHR2の役割を、モルフォリノアンチセンスを用いた遺伝子ノックダウン法で検討した。得られた結果より、PXRはセルフアップレギュレーションされること、ならびにPXRはAHR2・CYP1/2/3分子種の転写調節に、AHR2はPXR・CYP1/2/3分子種の転写調節にそれぞれ関与することが明らかとなった。こうした結果は、PXR・AHR2シグナル伝達系の間に双方向性のクロストークが存在することを示唆している。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Report

(1 results)
  • 2014 Annual Research Report
  • Research Products

    (12 results)

All 2015 2014

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Acknowledgement Compliant: 1 results) Presentation (9 results) (of which Invited: 2 results)

  • [Journal Article] Role of pregnane X receptor and aryl hydrocarbon receptor in transcriptional regulation of pxr, CYP2, and CYP3 genes in developing zebrafish.2015

    • Author(s)
      Kubota, A., Goldstone, J.V., Lemaire, B., Takata, M., Woodin, B.R., Stegeman, J.J.
    • Journal Title

      Toxicological Sciences

      Volume: 143 Issue: 2 Pages: 398-407

    • DOI

      10.1093/toxsci/kfu240

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enzymatic characterization of in vitro-expressed Baikal seal cytochrome P450 (CYP) 1A1, 1A2, and 1B1: Implication of low metabolic potential of CYP1A2 uniquely evolved in aquatic mammals2015

    • Author(s)
      Iwata H, Yamaguchi K, Takeshita Y, Kubota A, Hirakawa S, Isobe T, Hirano M, Kim EY.
    • Journal Title

      Aquat Toxicol

      Volume: 162 Pages: 138-151

    • DOI

      10.1016/j.aquatox.2015.03.010

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Involvement of COX2-thromboxane pathway in TCDD-induced precardiac edema in developing zebrafish2014

    • Author(s)
      Teraoka H, Okuno Y, Nijoukubo D, Yamakoshi A, Peterson R, Stegeman JJ, Kitazawa T, Hiraga T. and Kubota A.
    • Journal Title

      Auat Toxicol.

      Volume: 154 Pages: 19-26

    • DOI

      10.1016/j.aquatox.2014.04.025

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] The PXR "gene battery" in zebrafish2015

    • Author(s)
      Goldstone, J.V., Harbeitner, R., Kubota, A., Lemaire, B., Stegeman, J.J.
    • Organizer
      18th International symposium on Pollutant Responses in Marine Organisms (PRIMO18)
    • Place of Presentation
      ノルウェー・トロンハイム
    • Year and Date
      2015-05-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Reciprocal cross-talk between PXR and AHR2 signaling in response to their putative agonists in developing zebrafish2015

    • Author(s)
      Kubota, A., Goldstone, J.V., Lemaire, B., Takata, M., Woodin, B.R., Stegeman, J.J.
    • Organizer
      18th International symposium on Pollutant Responses in Marine Organisms (PRIMO18)
    • Place of Presentation
      ノルウェー・トロンハイム
    • Year and Date
      2015-05-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Differences in responses to putative PXR agonists associated with PXR alleles in zebrafish embryos2015

    • Author(s)
      Kubota, A., Goldstone, J.V., Lemaire, B., Harbeitner, R., Salay, L., Stegeman, J.J.
    • Organizer
      18th International symposium on Pollutant Responses in Marine Organisms (PRIMO18)
    • Place of Presentation
      ノルウェー・トロンハイム
    • Year and Date
      2015-05-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Transcriptomic effects of ortho-PCBs on developing zebrafish2015

    • Author(s)
      Goldstone, J.V., Lemaire, B., Kubota, A., Stegeman, J.J.
    • Organizer
      54th Annual Meeting of Society of Toxicology
    • Place of Presentation
      米国・サンディエゴ
    • Year and Date
      2015-03-26
    • Related Report
      2014 Annual Research Report
  • [Presentation] Zebrafish PXR inhibition in vivo by human PXR agonists2015

    • Author(s)
      Stegeman, J.J., Lemaire, B., Kubota, A., Harbeitner, R., Goldstone, J.V.
    • Organizer
      54th Annual Meeting of Society of Toxicology
    • Place of Presentation
      米国・サンディエゴ
    • Year and Date
      2015-03-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Cytochrome P450 20A1: Diversity and potential functions of a major orphan P450s2015

    • Author(s)
      Lemaire, B., Goldstone, J.V., Lamb, D., Kubota, A., Nelson, D., Stegeman, J.J.
    • Organizer
      54th Annual Meeting of Society of Toxicology
    • Place of Presentation
      米国・サンディエゴ
    • Year and Date
      2015-03-23
    • Related Report
      2014 Annual Research Report
  • [Presentation] Regulation of CYP genes by AHR and PXR in zebrafish2015

    • Author(s)
      Kubota, A.
    • Organizer
      Leading symposium on molecular targets of environmental chemicals
    • Place of Presentation
      北海道大学
    • Year and Date
      2015-02-10
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] PXR and AHR both are involved in transcriptional regulation of pxr, CYP2 and CYP3 genes in developing zebrafish2014

    • Author(s)
      Kubota, A., Goldstone, J.V., Lemaire, B., Stegeman, J.J.
    • Organizer
      環境ホルモン学会第17回研究発表会
    • Place of Presentation
      東京大学
    • Year and Date
      2014-12-10
    • Related Report
      2014 Annual Research Report
  • [Presentation] Studies on environmental toxicology with zebrafish: Toward understanding characteristics and molecular mechanisms of developmental toxicity of chemicals2014

    • Author(s)
      Kubota, A., Teraoka, H., Stegeman, J.J.
    • Organizer
      7th International Conference on Environmental Health Science
    • Place of Presentation
      韓国・ソウル
    • Year and Date
      2014-10-29
    • Related Report
      2014 Annual Research Report
    • Invited

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Published: 2014-09-09   Modified: 2016-06-01  

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