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位置固有な四肢軟骨成長を制御するHox遺伝子の標的遺伝子の分離同定

Publicly Offered Research

Project AreaMolecular mechanisms underlying reconstruction of 3D structers during regeneration
Project/Area Number 25124703
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionNagoya University

Principal Investigator

黒岩 厚  名古屋大学, 理学(系)研究科(研究院), 教授 (20134611)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000)
Fiscal Year 2014: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
KeywordsHox遺伝子 / 肢芽 / Hoxa13 / 遺伝子改変マウス / Hox標的遺伝子 / 四肢骨形成 / 四肢形成 / 軟骨形成 / ChIP-SEQ
Outline of Annual Research Achievements

(1)ChIP-SEQによる肢芽におけるHox標的遺伝子の同定
E12.5マウス胚肢芽自脚と軛脚領域を用いたHoxa13結合遺伝子を同定するChIP-SEQの結果が、共同研究において得られた。これによりHoxa13が発現する自脚領域に特異的なHoxa13結合ピークが数多く同定された。これらの中には、自脚でHoxa13に依存的に発現が抑制されると推定されるHoxa11のゲノム配列が確認され、ChIP-SEQが成功していることが示された。更に当研究室でKoxa13,Hoxd11-13ダブルKOマウス胚自脚で発現変動することが見いだされているFzd2, Sulf1, Gja3を含むいくつかの遺伝子についても自脚特異的ピークが確認され、これらが直接の標的遺伝子である可能性が示された。自脚パターン形成期であるE11.0でもChIPを行い、E12.5でHoxa13の標的候補として同定された複数の遺伝子について、Q-PCRで濃縮が確認され、現在ChIP-SEQのデータ解析を行っている。軛脚で特異的に発現するHoxa11についても、ChIP-SEQに使用できる抗体の選定を終え、Q-PCRで標的と推定されるFgf10の肢芽間充織エンハンサーの濃縮が確認される段階に到達し、近日中にChIP-SEQの段階に移行する。
(2)Hoxa13発現領域における標的遺伝子機能解析技術の開発
Hoxa-13:CreERT2-IRES venus SV40pAマウスの作成がゲノム解析で完了したことが判明し、次いでこの個体の胚でvenusがHoxa13と同じ発現をする事を確認した。このマウスとCAG loxP CAT loxP LacZマウスを交配して胚でのLacZ発現を解析した。誘導因子タモキシフェンの投与時のみHoxa13発現組織と同じ組織でLacZ発現が確認され、目的マウスの作成が成功したことが判明した。

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

(2 results)
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • Research Products

    (9 results)

All 2014 2013

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 3 results) Presentation (5 results) Book (1 results)

  • [Journal Article] Etv1 and Ewsr1 cooperatively regulate limb mesenchymal Fgf10 expression in response to apical ectodermal ridge-derived fibroblast growth factor signal2014

    • Author(s)
      Y. Yamamoto-Shiraishi, H. Higuchi, S. Yamamoto, M. Hirano and A. Kuroiwa
    • Journal Title

      Dev Biol

      Volume: 394 Issue: 1 Pages: 181-90

    • DOI

      10.1016/j.ydbio.2014.07.022

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient embryonic culture method for the Japanese striped snake, Elaphe quadrivirgata, and its early developmental stages2014

    • Author(s)
      Y. Matsubara, A. Sakai, A. Kuroiwa and T. Suzuki
    • Journal Title

      Dev Growth Differ

      Volume: 56 Issue: 8 Pages: 573-82

    • DOI

      10.1111/dgd.12157

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Leucophores are similar to xanthophores in their specification and differentiation processes in medaka2014

    • Author(s)
      T. Kimura, Y. Nagao, H. Hashimoto, Y. Yamamoto-Shiraishi, S. Yamamoto, T. Yabe, S. Takada, M. Kinoshita, A. Kuroiwa and K. Naruse
    • Journal Title

      Proc Natl Acad Sci U S A

      Volume: 111 Issue: 20 Pages: 1986767-1986767

    • DOI

      10.1073/pnas.1311254111

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Heterochrony in initiation of paraxial Gdf11 expression specifies unique hindlimb positioning in tetrapods.2014

    • Author(s)
      Yoshiyuki Matsubara, Ayumi Hattori, Yusuke Watanabe, Atsushi Sakai, Guojun Sheng, Toshihiko Ogura, Atsushi Kuroiwa, Takayuki Suzuki
    • Organizer
      47th Annual Meeting for the Japanese Society of Developmental Biologists
    • Place of Presentation
      ウィンクあいち
    • Year and Date
      2014-05-27 – 2014-05-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] The quail polydactyly mutant, HMM, shows inactivation of SHH signaling.2014

    • Author(s)
      Kazuki Kawamura, Yoshiyuki Matsubara, Mikiharu Nakano, Masaoki Tsudzuki, Youichi Matsuda, Atsushi Kuroiwa, Takayuki Suzuki.
    • Organizer
      47th Annual Meeting for the Japanese Society of Developmental Biologists
    • Place of Presentation
      ウィンクあいち
    • Year and Date
      2014-05-27 – 2014-05-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] Analysis of the function of R31 limb specific enhancer of Fgf10.2014

    • Author(s)
      Chisa Ando, Yu-hei Kohara, YO-ichi Yamamoto-Shiraishi, Atsushi Kuroiwa
    • Organizer
      47th Annual Meeting for the Japanese Society of Developmental Biologists
    • Place of Presentation
      ウィンクあいち
    • Year and Date
      2014-05-27 – 2014-05-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] Transcriptional regulation mechanisms of mesenchymal Fibrobrast Growth Factor 10 expression in responce to the AER signaling2013

    • Author(s)
      Yo-Ich Yamamoto-Shiraishi, Hiroaki Higuchi, Shigeki Yamamoto, Mie Hirano, Atsushi Kuroiwa
    • Organizer
      46th Annual Meeting for the Japanese Society of Developmental Biologists
    • Place of Presentation
      Kunubuki Messe(Matsue-shi, Shimane-ken)
    • Related Report
      2013 Annual Research Report
  • [Presentation] Hox and Wnt signaling cooperatively control Fgf10 expression in the limb field2013

    • Author(s)
      Taishi Ueta, Hiroyuki Inaba, Takehiro Hitokoto, Yo-Ichi Shiraishi, Takayuki Suzuki, Atsushi Kuroiwa
    • Organizer
      46th Annual Meeting for the Japanese Society of Developmental Biologists
    • Place of Presentation
      Kunubuki Messe(Matsue-shi, Shimane-ken)
    • Related Report
      2013 Annual Research Report
  • [Book] New Principles in Developmental Processes2014

    • Author(s)
      Hisato Kondoh, Atsushi Kuroiwa Edt
    • Total Pages
      321
    • Publisher
      Springer
    • Related Report
      2013 Annual Research Report

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Published: 2013-05-15   Modified: 2019-07-29  

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