Function analysis of TBT-binding protein

• Project/Area Number: 18H02280
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 40040:Aquatic life science-related
• Research Institution: Kyushu University
• Principal Investigator: Oshima Yuji 九州大学, 農学研究院, 教授 (70176874)
• Co-Investigator(Kenkyū-buntansha): 荻野 由紀子 九州大学, 農学研究院, 准教授 (00404343) ; 鵜木 陽子 (加藤陽子) 九州大学, 農学研究院, 技術専門職員 (10380560) ; 太田 耕平 九州大学, 農学研究院, 准教授 (10585764) ; 北野 健 熊本大学, 大学院先端科学研究部(理), 教授 (40336219) ; 島崎 洋平 九州大学, 農学研究院, 准教授 (40363329) ; 杣本 智軌 九州大学, 農学研究院, 准教授 (40403993)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2020: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2019: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2018: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: 魚 / リポカリン / 性決定 / 解毒 / 生体防御 / 魚リポカリン / TBT-bp / 遺伝子編集 / 高病性 / 白点虫感染 / 魚類リポカリン / ストレス応答 / 異物結合能 / 感染防御 / ストレス / AGP / アルファ1酸性グリコプロテイン / リポカリンタンパク質 / オス化 / ノックアウト / TBT-binding protein

Outline of Final Research Achievements

This study was performed to analyze the function of TBT-bps using knock out (KO) medaka, Oryzias latipes. First, the growth and sex differentiation were compared in TBT-bp2 KO medaka fish. As results, the no difference was observed in growth of TBT-bp2 medaka. However, this strain female medaka change to functional male by exposure of cortisol. TBT-bp2 might be indirectly related to sex determination. In second, we successfully knock in GFP gene to TBT-bp2, and generate TBT-bp2KI homo medaka. Using this medaka , expression of TBT-bp2 in liver and its circulation in blood stream were detected by GFP luminesce. TBT-bp2KO medaka were exposed to toxic chemical or parasite and their tolerance was observed. The decrease of survival of chemical or parasite were observed. Thus, TBT-bps might be involved in detoxification process and biological defenses. In addition, expression levels of TBT-bp1 and 2 of aquacultured fish were affected by their maintain conditions.

Academic Significance and Societal Importance of the Research Achievements

本研究により魚類血中で高濃度に存在するがその役割が未解明であった魚リポカリン=TBT-bpsの機能が解明され，生体防御，解毒，性決定に関与していることがわかった．TBT-bpsは魚類の性分化，代謝，恒常性の維持に極めて重要であり，その変動を調べることで健康管理のバイオマーカーとしての活用が期待される．さらに本プロジェクトで作製したTBT-bp2KI, KOメダカを用いることでよりさらなる機能解析が進むと期待される．

Research Products

• [Journal Article] Production of a tributyltin-binding protein 2 knockout mutant strain of Japanese medaka, Oryzias latipes (2020)
  • Author(s): Kato-Unoki Yoko、Takai Yuki、Nagano Yosuke、Matsunaga Satoshi、Enoki Shintaro、Takamura Takumi、Kim Sangwan、Kinoshita Masato、Kitano Takeshi、Shimasaki Yohei、Oshima Yuji
  • Journal Title: Marine Pollution Bulletin Volume: 160 Pages: 111601-111601
  • DOI: 10.1016/j.marpolbul.2020.111601
  • Peer Reviewed
• [Journal Article] Detoxification roles of tributyltin-binding protein type 2 in Japanese medaka (Oryzias latipes) exposed to tributyltin. (2020)
  • Author(s): Qiu, X., Takamura, T., Enoki, S., Kato-Unoki, Y., Takai, Y., Nagano, Y., Kinoshita, M., Kitano, T., Shimasaki, Y., Oshima, Y.,
  • Journal Title: Mar. Pollut. Bull. 159, 111445 Volume: 159 Pages: 111445-111445
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Establishment of a Japanese medaka (Oryzias latipes) transgenic line expressing Takifugu rubripes pufferfish saxitoxin and tetrodotoxin binding protein 1, and evaluation of tributyltin toxicity via in ovo nanoinjection. (2020)
  • Author(s): Takai, Y., Mizoguchi, N., Kinoshita, M., Qiu, X., Shimasaki, Y., Oshima, Y
  • Journal Title: Comp. Biochem. Physiol. C Pharmacol. Toxicol. Endocrinol. Volume: 234 Pages: 108785-108785
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transcriptome analysis of medaka (Oryzias latipes) exposed to tributyltin. (2020)
  • Author(s): Takai, Y., Takamura, T., Enoki, S., Sato, M., Kato-Unoki, Y., Qiu, X., Shimasaki, Y., Oshima, Y.,
  • Journal Title: Jap. J. Toxicol. Environ. Health Volume: 23 Pages: 10-21
  • NAID: 130007879233
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 魚類リポカリンタンパク質 TBT-bp2破壊メダカにおける白点虫感染機構の解析 (2021)
  • Author(s): 高井優生, 助田将樹, 高村匠, 長野陽介, 島崎洋平, 杣本智軌, 大嶋雄治
  • Organizer: 令和２年度日本水産学会春季大会
• [Presentation] 魚類リポカリンTBT-bp2-GFPノックインメダカの作成. (2021)
  • Author(s): 榎眞太郎, 鵜木（加藤）洋子, 荻野由紀子, 島崎洋平,大嶋雄治
  • Organizer: 令和２年度日本水産学会春季大会
• [Presentation] Toxicity of tributyltin and triclosan in tributyltin-binding protein 2 knocked-out medaka, Oryzias latipes. (2019)
  • Author(s): Y. Oshima, T. Takamura, S. Enoki, Y. Kato-Unoki, Y. Nagano, M. Kinoshit, T. Kitano, I. J. Kang, X. Qiu, Y. Shimasaki
  • Organizer: SETAC North America 40th Annual Meeting
  • Int'l Joint Research
• [Presentation] Toxicity of tributyltin and triclosan in tributyltin-binding protein 2 knocked-out medaka, Oryzias latipe (2019)
  • Author(s): Y. Oshima, T. Takamura, S. Enoki, Y. Kato-Unoki, Y. Nagano, M. Kinoshita, T. Kitano, IJ. Kang, X. Qiu, Y. Shimasaki.
  • Organizer: 9th International Conference on Marine Pollution and Ecotoxicology
  • Int'l Joint Research
• [Presentation] ノックアウトメダカを用いたトリブチルスズ 結合タンパク質のトリクロサン毒性低減機能解析 (2019)
  • Author(s): 高村 匠・榎 眞太郎・上野 雄・鵜木(加藤)陽子(九大院農)・邱 旭春・島崎洋平・大嶋雄治
  • Organizer: Ｈ３１年度日本水産学会春季大会
• [Presentation] TOXICITY OF TRIBUTYLTIN AND TRICLOSAN IN TRIBUTYLTIN-BINDING PROTEIN 2 KNOCKED-OUT MUTANT STRAIN OF JAPANESE MEDAKA, ORYZIAS LATIPES (2019)
  • Author(s): Yuji Oshima, Takumi Takamura, Shintaro Enoki, Yoko Kato-Unoki, Yosuke Nagano, Masato Kinoshita, Takeshi Kitano, Ik Joon Kang, Xuchun Qiu, Yohei Shimasaki
  • Organizer: 9th International Conference on Marine Pollution and Ecotoxicology
  • Int'l Joint Research