Building new foundation for biological sciences based on transposon-mediated gene trapping

2017 Fiscal Year Annual Research Report

• Project/Area Number: 15H02370
• Research Institution: National Institute of Genetics
• Principal Investigator: 川上 浩一 国立遺伝学研究所, 個体遺伝研究系, 教授 (70195048)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ゲノム / 遺伝学 / 遺伝子 / 神経科学 / 発生・分化 / ゼブラフィッシュ / 遺伝子トラップ

Outline of Annual Research Achievements

（１）ゼブラフィッシュにおいて、Tol2を用いた遺伝子トラップコンストラクトをもつプラスミドDNAを、転移酵素mRNAとともに受精卵にインジェクションし、F1胚をスクリーニングすることにより、細胞・組織・器官特異的に改変型酵母転写因子Gal4FFを発現するトランスジェニックフィッシュを新規に100系統作製した。サザンブロッティング、インバースPCRによりこれらにおけるTol2挿入部位近傍のゲノムDNAの解析を行い、挿入部位を決定した。それら発現情報、遺伝子情報をとりこんだデータベースの整備を行った。
（２）細胞・組織・器官特異的Gal4FF発現トランスジェニックフィッシュを用い、以下のことを明らかにした。血液脳関門形成に関わる新規細胞群の発見（米国NIH、Weinstein研究との共同研究）。小脳顆粒細胞が恐怖条件付け学習の回復期に機能することを発見（名古屋大日比研との共同研究）。稚魚の逃避行動に関わるグリシンリセプターのサブタイプを同定（豪州サンシャインコースト大Harvey研との共同研究）。脊索障害の修復の際に機能する新規細胞群を発見（英国エジンバラ大Patton研との共同研究）。
（３）Gal4FF発現トランスジェニックゼブラフィッシュのスクリーニングを行い、前視蓋あるいは視床下部下葉の特定の脳神経回路、および終脳の特定の神経細胞で、Gal4FFを発現するトランスジェニックフィッシュを同定した。これらを基にして、カルシウムイメージングおよび機能阻害実験を行い、エサなどの視覚刺激が食欲の中枢である視床下部下葉を活性化する神経回路を発見した（Muto et al. Nat Comm 2017）。また、恐怖条件付け学習に必須な終脳の特定の神経細胞を明らかにした(Lal et al. BMC Biol 2018)。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] National Institute of Health/Salk Institute for Biological Studies(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: National Institute of Health/Salk Institute for Biological Studies
• [Int'l Joint Research] University of the Sunshine Coast/The University of Queensland(オーストラリア)
  • Country Name: AUSTRALIA
  • Counterpart Institution: University of the Sunshine Coast/The University of Queensland
• [Int'l Joint Research] UCL School of Pharmacy/University of Edinburgh(英国)
  • Country Name: UNITED KINGDOM
  • Counterpart Institution: UCL School of Pharmacy/University of Edinburgh
• [Int'l Joint Research] Hubrecht Institute(オランダ)
  • Country Name: NETHERLANDS
  • Counterpart Institution: Hubrecht Institute
• [Int'l Joint Research] Friedrich-Schiller-University(ドイツ)
  • Country Name: GERMANY
  • Counterpart Institution: Friedrich-Schiller-University
• [Int'l Joint Research]
  • # of Other Countries: 1
• [Journal Article] Structure/Function Studies of the α4 Subunit Reveal Evolutionary Loss of a GlyR Subtype Involved in Startle and Escape Responses (2018)
  • Author(s): Leacock, S., Syed, P., James, V.M., Bode, A., Kawakami, K., Keramidas, A., Suster, M., Lynch, J.W., and Harvey, R.J.
  • Journal Title: Frontiers in Molecular Neuroscience Volume: 11 Pages: 23
  • DOI: doi.org/10.3389/fnmol.2018.00023
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Wilms Tumor 1b defines a wound-specific sheath cell subpopulation associated with notochord repair (2018)
  • Author(s): Lopez-Baez, J.C., Simpson, D.J., Forero, L.L., Zeng, Z., Brunsdon, H., Salzano, A., Brombin, A., Wyatt, C., Rybski, W., Huitema, L.FA., Dale, R.M., Kawakami, K., Englert, C., Chandra, T., Schulte-Merker, S., Hastie, N.D., and Patton, E.E.
  • Journal Title: eLife Volume: 7 Pages: e30657
  • DOI: 10.7554/eLife.30657
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of a neuronal population in the telencephalon essential for fear conditioning in zebrafish (2018)
  • Author(s): Lal, P., Tanabe, H., Suster, M.L., Ailani, D., Kotani, Y., Muto, A., Itoh, M., Iwasaki, M., Wada, H., Yaksi, E., and Kawakami, K.
  • Journal Title: BMC Biology Volume: 16 Pages: 45
  • DOI: 10.1186/s12915-018-0502-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Activation of the hypothalamic feeding centre upon visual prey detection (2017)
  • Author(s): Muto, A., Lal, P., Ailani, D., Abe, G., Itoh, M., and Kawakami, K.
  • Journal Title: Nature Communications Volume: 8 Pages: 15029
  • DOI: doi: 10.1038/ncomms15029
  • Peer Reviewed / Open Access
• [Journal Article] A novel perivascular cell population in the zebrafish brain (2017)
  • Author(s): Galanternik, M.V., Castranova, D., Gore, A.V., Blewett, N.H., Jung, H.M., Stratton, A.N., Kirby, M.R., Iben, J., Miller, M.F., Kawakami, K., Maraia, R.J., and Weinstein, B.M.
  • Journal Title: eLife Volume: 6 Pages: e24369
  • DOI: 10.7554/eLife.24369
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Analysis of transcription factors expressed at the anterior mouse limb bud (2017)
  • Author(s): Yokoyama, S., Furukawa, S., Kitada, S., Mori, M., Saito, T., Kawakami, K., Belmonte, J. C. I., Kawakami, Y., Ito, Y., Sato, T., and Asahara, H.
  • Journal Title: PLoS ONE Volume: 12 Pages: e0175673
  • DOI: 10.1371/journal.pone.0175673
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Activin-A enhances mTOR signaling to promote aberrant chondrogenesis in fibrodysplasia ossificans progressiva (2017)
  • Author(s): Hino, K., Horigome, K., Nishio, M., Komura, S., Nagata, S., Zhao, C., Jin, Y., Kawakami, K., Yamada, Y., Ohta, A., Toguchida, J., and Ikeya, M.
  • Journal Title: The Journal of Clinical Investigation Volume: 127 Pages: 3339-3352
  • DOI: 10.1172/JCI93521
  • Peer Reviewed / Open Access
• [Journal Article] Preface to Vertebrate Brains: evolution, structures and functions (2017)
  • Author(s): Kawakami, K., and Murakami, Y.
  • Journal Title: Development, Growth and Differentiation Volume: 59 Pages: 160-162
  • DOI: 10.1111/dgd.12375
  • Open Access
• [Journal Article] Transposons as tools for functional genomics in vertebrate models (2017)
  • Author(s): Kawakami, K., Largaespada, D.A., and Ivics, Z.
  • Journal Title: Trends in Genetics Volume: 33 Pages: 7840801
  • DOI: doi.org/10.1016/j.tig.2017.07.006
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Granule cells control recovery from classical conditioned fear responses in the zebrafish cerebellum (2017)
  • Author(s): Matsuda, K., Yoshida, M., Kawakami, K., Hibi, M., and Shimizu. T.
  • Journal Title: Scientific reports Volume: 7 Pages: 11865
  • DOI: 10.1038/s41598-017-10794-0
  • Peer Reviewed / Open Access
• [Presentation] The amygdalar and hippocampal functions in zebrafish (2018)
  • Author(s): Koichi Kawakami
  • Organizer: The 5th European Zebrafish PI Meeting
  • Int'l Joint Research
• [Presentation] The neuronal circuitry for prey capture in zebrafish: from visual stimuli to the hypothalamic feeding center (2017)
  • Author(s): Kawakami, K.
  • Organizer: The 10th Zebrafish Disease Models Conference
  • Int'l Joint Research / Invited
• [Presentation] Enhancer trapping and calcium imaging reveal functional neuronal circuitries essential for fish behaviors (2017)
  • Author(s): Kawakami, K.
  • Organizer: 2nd Malaysia Zebrafish Disease Model Workshop
  • Int'l Joint Research / Invited
• [Presentation] Enhancer trapping and calcium imaging reveal a functional neuronal circuit essential for the prey capture behavior: from visual stimuli to the hypothalamic feeding center (2017)
  • Author(s): Kawakami, K.
  • Organizer: The 5th Zebrafish Research Conference of China
  • Int'l Joint Research / Invited
• [Presentation] The amygdalar and hippocampal functions in zebrafish The 2017 workshop on Zebrafish (2017)
  • Author(s): Kawakami, K.
  • Organizer: Neural Circuits and Behavior
  • Int'l Joint Research / Invited
• [Remarks] gene trap & enhancer trap transgenic zebrafish
  • URL: http://kawakami.lab.nig.ac.jp/ztrap/