トランスポゾンを用いた方法論に基づく脊椎動物フェノーム解析の基盤形成

2013 Fiscal Year Annual Research Report

• Project/Area Number: 23241063
• 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): 2011-04-01 – 2015-03-31
• Keywords: ゲノム / 遺伝学 / 遺伝子 / 神経科学 / 発生・分化

Research Abstract

（１）モデル脊椎動物ゼブラフィッシュにおいて、Gal4を組み込んだ遺伝子トラップトランスポゾンコンストラクトをゲノムにランダムに組み込ませたトランスジェニックフィッシュから、細胞・組織・器官特異的にGal4を発現するトランスジェニックフィッシュを選別する大規模スクリーニングを行い、平成25年度、新規100系統を作製することに成功した。これらをかけあわせ単一挿入をもつ系統を樹立し、インバースPCR等分子生物学的解析を行うことにより、トランスポゾン挿入部位を決定した。
（２）本研究により作製してきたトランスジェニックフィッシュを用いて、国内外の研究者と初期発生、器官形成、神経科学に関する共同研究を行った。ゼブラフィッシュの感覚器官である感丘の分裂・増殖における神経細胞からのwntシグナルの重要性、WntとそのインヒビターDkkのフィードバックによる器官サイズの制御機構（和田博士との共同研究）、心臓弁形成におけるマイクロRNAによるターゲット遺伝子の発現制御（小椋博士との共同研究）、神経細胞を染め分けるトランスジェニックゼブラフィッシュ系統の開発（東島博士との共同研究）、運動神経の軸索伸長における血管からの成長因子の役割（望月博士との共同研究）、嗅覚からのインプットによる脳左右半球非対称性の形成（澤本博士との共同研究）、発生過程の脳形成における転写因子Gbx2の役割（弥益博士との共同研究）、神経細胞移動におけるVangl2の役割（Chandrasekhar博士との共同研究）を明らかにした。
（３）脳の様々な領域でGal4を特異的に発現するトランスジェニックフィッシュ系統をUAS-神経毒素遺伝子トランスジェニックフィッシュ系統とかけあわせ、行動解析実験を行うことにより、恐怖条件付け学習に必要な脳領域の同定に成功した。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

申請時目標とした新規トランスジェニックフィッシュの年度作製数50系統を大幅に越えて新規系統の開発に成功した。またそれらに基づいて、我々自身の研究室および共同研究者の研究室から平成25年度中に関連論文計16報を発表することができたから。

Strategy for Future Research Activity

研究は当初の計画以上に進展しており、今後のさらなる発展を目指す。

Research Products

• [Journal Article] Targeted expression of a chimeric channelrhodopsin in zebrafish under regulation of Gal4-UAS system (2013)
  • Author(s): Umeda, K., Shoji, W., Sakai, S., Muto, A., Kawakami, K., Ishizuka, T., and Yawo, H.
  • Journal Title: Neuroscience Research Volume: 75 Pages: 69-75
  • DOI: 10.1016/j.neures.2012.08.010
  • Peer Reviewed
• [Journal Article] Innervation is required for sense organ development in the lateral line system of adult zebrafish (2013)
  • Author(s): Wada, H., Dambly-Chaudiere, C., Kawakami, K., and Ghysen, A.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 110 Pages: 5659-5664
  • DOI: 10.1073/pnas.1214004110
  • Peer Reviewed
• [Journal Article] Transcription factors interfering with dedifferentiation induce cell type-specific transcriptional profiles (2013)
  • Author(s): Hikichi, T., Matoba, R., Ikeda, T., Watanabe, A., Yamamoto, T., Yoshitake, S., Tamura-Nakano, M., Kimura, T., Kamon, M., Shimura, M., Kawakami, K., Okuda, A., Okochi, H., Inoue, T., Suzuki, A., and Masui, S.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 110 Pages: 6412-6417
  • DOI: 10.1073/pnas.1220200110
  • Peer Reviewed
• [Journal Article] Differential role for transcription factor Oct4 nucleocytoplasmic dynamics in somatic cell reprogramming and self-renewal of embryonic stem cells (2013)
  • Author(s): Oka, M., Moriyama, T., Asally, M., Kawakami, K., and Yoneda, Y.
  • Journal Title: The Journal of Biological Chemistry Volume: 288 Pages: 15085-15097
  • DOI: 10.1074/jbc.M112.448837
  • Peer Reviewed
• [Journal Article] Interhemispheric asymmetry of olfactory input-dependent neuronal specification in the adult brain (2013)
  • Author(s): Kishimoto, N., Asakawa, K., Madelaine, R., Blader, P., Kawakami, K., and Sawamoto, K.
  • Journal Title: Nature Neuroscience Volume: 16 Pages: 884-888
  • DOI: 10.1038/nn.3409
  • Peer Reviewed
• [Journal Article] Cellular dissection of the spinal cord motor column by BAC transgenesis and gene trapping in zebrafish (2013)
  • Author(s): Asakawa, K., Abe, G., and Kawakami, K.
  • Journal Title: Frontiers in Neural Circuits Volume: 7 Pages: 100
  • DOI: 10.3389/fncir.2013.00100
  • Peer Reviewed
• [Journal Article] Prey capture in zebrafish larvae serves as a model to study cognitive functions (2013)
  • Author(s): Muto, A., and Kawakami, K.
  • Journal Title: Frontiers in Neural Circuits Volume: 7 Pages: 110
  • DOI: 10.3389/fncir.2013.00110
  • Peer Reviewed
• [Journal Article] Haemodynamically dependent valvulogenesis of zebrafish heart is mediated by flow-dependent expression of miR-21 (2013)
  • Author(s): Banjo, T., Grajcarek, J., Yoshino, D., Osada, H., Miyasaka, K.Y., Kida, Y.S., Ueki, Y., Nagayama, K., Kawakami, K., Matsumoto, T., Sato, M., and Ogura, T.
  • Journal Title: Nature Communications Volume: 4 Pages: 1978
  • DOI: 10.1038/ncomms2978
  • Peer Reviewed
• [Journal Article] Wnt/Dkk negative feedback regulates sensory organ size in zebrafish (2013)
  • Author(s): Wada, H., Ghysen, A., Asakawa, K., Abe, G., Ishitani, T., and Kawakami, K.
  • Journal Title: Current Biology Volume: 23 Pages: 1559-1565
  • DOI: 10.1016/j.cub.2013.06.035
  • Peer Reviewed
• [Journal Article] Development of cerebellar neurons and glias revealed by in utero electroporation: Golgi-like labeling of cerebellar neurons and glias (2013)
  • Author(s): Kita, Y., Kawakami, K., Takahashi, Y. and Murakami, F.
  • Journal Title: PLoS One Volume: 8 Pages: e70091
  • DOI: 10.1371/journal.pone.0070091
  • Peer Reviewed
• [Journal Article] Transgenic tools to characterize neuronal properties of discrete populations of zebrafish neurons (2013)
  • Author(s): Satou, C., Kimura, Y., Hirata, H., Suster, M.L., Kawakami, K., and Higashijima, S.-i.
  • Journal Title: Development Volume: 140 Pages: 3927-3931
  • DOI: 10.1242/dev.099531
  • Peer Reviewed
• [Journal Article] The parallel growth of motoneuron axons with the dorsal aorta depends on Vegfc/Vegfr3 signaling in zebrafish (2013)
  • Author(s): Kwon, H.-B., Fukuhara, S., Asakawa, K., Ando, K., Kashiwada, T., Kawakami, K., Hibi, M., Kwon, Y.-G., Kim, K.-W., Alitalo, K., and Mochizuki, N.
  • Journal Title: Development Volume: 140 Pages: 4081-4090
  • DOI: 10.1242/dev.091702
  • Peer Reviewed
• [Journal Article] Gbx2 functions as a transcriptional repressor to regulate the specification and morphogenesis of the mid-hindbrain junction in a dosage- and stage-dependent manner (2013)
  • Author(s): Nakayama, Y., Kikuta, H., Kanai, M., Yoshikawa, K., Kawamura, A., Kobayashi, K., Wang, Z., Khan, A., Kawakami, K., and Yamasu, K.
  • Journal Title: Mechanisms of Development Volume: 130 Pages: 532-552
  • DOI: 10.1016/j.mod.2013.07.004
  • Peer Reviewed
• [Journal Article] The PCP protein Vangl2 regulates migration of hindbrain motor neurons by acting in floor plate cells, and independently of cilia function (2013)
  • Author(s): Sittaramane, V., Pan, X., Glasco, D.M., Huang, P., Gurung, S., Bock, A., Li, S., Wang, H., Kawakami, K., Matise, M.P., and Chandrasekhar, A.
  • Journal Title: Developmental Biology Volume: 382 Pages: 400-412
  • DOI: 10.1016/j.ydbio.2013.08.017
  • Peer Reviewed
• [Journal Article] トランスジェニックフィッシュが拓く新しい生物学 (2013)
  • Author(s): 川上浩一
  • Journal Title: 細胞工学 Volume: 32 Pages: 898-903
• [Journal Article] ゼブラフィッシュを用いた脳神経活動のリアルタイムイメージング (2013)
  • Author(s): 武藤彩, 川上浩一
  • Journal Title: 細胞工学 Volume: 32 Pages: 700-701
• [Presentation] Genetic dissection of the zebrafish brain by the Gal4-UAS method (2014)
  • Author(s): Kawakami, K.
  • Organizer: 国際高等研究所研究プロジェクト「ゲノム工学とイメージングサイエンスに基づく生命システム研究の新展開」第2回研究会
  • Place of Presentation: 京都
  • Year and Date: 20140227-20140228
• [Presentation] Genetic dissection of the adult zebrafish brain by the Gal4-UAS method (2014)
  • Author(s): Kawakami, K.
  • Organizer: 6th Asia Oceania Zebrafish Meeting
  • Place of Presentation: 香港
  • Year and Date: 20140119-20140122
  • Invited
• [Presentation] Genetic dissection of the adult zebrafish brain by the Gal4-UAS method (2013)
  • Author(s): Kawakami, K., Lal, P., and Itoh, M.
  • Organizer: The 36th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: 神戸
  • Year and Date: 20131203-20131206
• [Presentation] Development of functional cerebellar neural circuitry in zebrafish (2013)
  • Author(s): Hibi, M., Takeuchi, M., Yamaguchi, S., Matsuda, K., Yonemura, S., Asakawa, K., Takada, S., Kawakami, K., and Shimizu, T.
  • Organizer: The 36th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: 神戸
  • Year and Date: 20131203-20131206
• [Presentation] Directional control of locomotion in the touch-evoked escape behavior in zebrafish (2013)
  • Author(s): Asakawa, K.,Abe, G.,Muto, A.,Kawakami, K.
  • Organizer: Neuroscience 2013
  • Place of Presentation: San Diego, USA
  • Year and Date: 20131109-20131113
• [Presentation] ZeBrain: A database of the Gal4 expression patterns in the adult zebrafish brain (2013)
  • Author(s): Kawakami, K.
  • Organizer: Jenelia workshop on zebrafish genetics, transgenesis, and systems biology
  • Place of Presentation: Ashburn, USA
  • Year and Date: 20131104-20131105
  • Invited
• [Presentation] A Gal4FF driver transgenic fish resource for developmental biology and neuroscience (2013)
  • Author(s): Kawakami, K.
  • Organizer: The 3rd Chinese Zebrafish Conference
  • Place of Presentation: 蘇州, 中国
  • Year and Date: 20131011-20131014
  • Invited
• [Presentation] Development of cerebellar granule cell axons in zebrafish (2013)
  • Author(s): Yamaguchi, S., Takeuchi, M., Asakawa, K., Kawakami, K., Shimizu, T., and Hibi, M.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Basement membranedependent control of axogenesis of cerebellar granule cells (2013)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Shimizu, T., and Hibi, M.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Functional analysis of zebrafish cerebellar neural circuitry in classical conditioning (2013)
  • Author(s): Matsuda, K., Takeuchi, M., Asakawa, K., Yoshida, M., Ohkura, M., Miyasaka, N., Yoshihara, Y., Nakai, J., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Development of the lateral line canal system in zebrafish (2013)
  • Author(s): Wada, H., and Kawakami, K.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Differential actions of Fgf signaling controls blastema formation and regeneration in zebrafish (2013)
  • Author(s): Horita, N., Abe, G., Kawakami, K., Kudo, A., and Kawakami, A.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Identification of bone stem cells during growth and regeneration of endo- and exo-skeletons in zebrafish (2013)
  • Author(s): Ando, K., Age, G., Kudo, A., Kawakami, K., and Kawakami, A.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Serotonin regulation by the habenula is essential for adaptive problem solving in zebrafish (2013)
  • Author(s): Amo, R., Agetsuma, M., Kinoshita, M., Fredes, F., Shiraki, T., Aoki, T., Aoki, R., Yamazaki, M., Higashijima, S.-i., Matsuda, M., Suster, M.L., Kawakami, K., Ohshima, T., Aizawa, H., and Okamoto, H.
  • Organizer: The 19th Japanese Medaka and Zebrafish Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20130920-20130921
• [Presentation] Tol2: forward and reverse genetics in zebrafish and transposition in S. cerevisiae (2013)
  • Author(s): Kawakami, K., and Iwasaki, M.
  • Organizer: Conference on Transposition and Genome Engineering 2013
  • Place of Presentation: Budapest, Hungary
  • Year and Date: 20130918-20130921
  • Invited
• [Presentation] Dissection of the spinal cord motor column by BAC transgenesis and gene trapping in zebrafish (2013)
  • Author(s): Asakawa, K., Abe, G., and Kawakami, K.
  • Organizer: 8th European Zebrafish Meeting
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 20130719-20130723
• [Presentation] Evolution of the neo-Y chromosome through co-option of Sox3 in a medaka-related fish (2013)
  • Author(s): Takehana, Y., Matsuda, M., Matsuyama, H., Myosho, T., Suster, M., Kawakami, K., Shin-i, T., Kohara, Y., Kuroki, Y., Toyoda, A., Fujiyama, A., Hamaguchi, S., Sakaizumi, M., and Naruse, K.
  • Organizer: 8th European Zebrafish Meeting
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 20130719-20130723
• [Presentation] A Gal4 driver resource for developmental biology and neuroscience in zebrafish (2013)
  • Author(s): Kawakami, K., Ailani, D., Asakawa, K., Tanabe, H., Lal, P., Miller, A.S., Muto, A., Wada, H., and Yoshino, A.
  • Organizer: 8th European Zebrafish Meeting
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 20130719-20130723
  • Invited
• [Presentation] ゼブラフィッシュ手綱核-縫線核経路は逃避学習行動を制御する (2013)
  • Author(s): 天羽龍之介, 揚妻正和, 木下雅恵, 白木利幸, 青木田鶴, 山崎昌子, 東島眞一, 松田勝, 川上浩一, 大島登志男, 相澤秀紀, 岡本仁
  • Organizer: 第36回日本神経科学大会
  • Place of Presentation: 京都
  • Year and Date: 20130620-20130623
• [Presentation] The dorsomedial telencephalon is essential for active avoidance response in zebrafish (2013)
  • Author(s): Lal, P., Hiratani, M., Suster, M.L., Kawakami, K.
  • Organizer: 第36回日本神経科学大会
  • Place of Presentation: 京都
  • Year and Date: 20130620-20130623
• [Presentation] Development of cerebellar granule cell axons in zebrafish (2013)
  • Author(s): Yamaguchi, S., Takeuchi, M., Asakawa, K., Kawakami, K., Shimizu, T., Hibi, M.
  • Organizer: 第46回日本発生生物学会
  • Place of Presentation: 松江
  • Year and Date: 20130528-20130531
• [Presentation] Development of the Trunk Lateral Line System through a Bone Remodeling Process in Zebrafish (2013)
  • Author(s): Wada, H., Kawakami, K.
  • Organizer: 第46回日本発生生物学会
  • Place of Presentation: 松江
  • Year and Date: 20130528-20130531
• [Presentation] Cellular dissection of the spinal cord motor column by BAC transgenesis and gene trapping in zebrafish (2013)
  • Author(s): Asakawa, K., Abe, G., Kawakami, K.
  • Organizer: 第46回日本発生生物学会
  • Place of Presentation: 松江
  • Year and Date: 20130528-20130531
• [Presentation] A Gal4 driver resource for developmental biology and neuroscience in zebrafish (2013)
  • Author(s): Kawakami, K., Ailani, D., Asakawa, K., Lal, P., Tanabe, H., Miller, A.S., Muto, A., Yoshino, A.
  • Organizer: 第46回日本発生生物学会
  • Place of Presentation: 松江
  • Year and Date: 20130528-20130531
• [Presentation] Basement membrane integrity is required for axogenesis of cerebellar granule cells (2013)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Shimizu, T., Hibi, M.
  • Organizer: 第46回日本発生生物学会
  • Place of Presentation: 松江
  • Year and Date: 20130528-20130528
• [Presentation] The Zebrafish VIPLa mutant is defective in activating the Mauthner cells in the escape behavior
  • Author(s): Asakawa, K.,and Kawakami, K.
  • Organizer: 第３２回日本糖質学会年会
  • Place of Presentation: 大阪
• [Presentation] Beyond a retrovirus: a quest with a transposon
  • Author(s): Kawakami, K.
  • Organizer: Nancy Hopkins Lab Reunion Symposium
  • Place of Presentation: Cambridge, MA, USA
  • Invited