Analysis of molecular mechanism in vertebrate brain regionalization

• Project/Area Number: 19570217
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Developmental biology
• Research Institution: The University of Tokyo (2008); National Institute of Advanced Industrial Science and Technology (2007)
• Principal Investigator: TATSUO Michiue The University of Tokyo, 大学院・総合文化研究科, 准教授 (10282724)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2007: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 形態形成 / 発生・分化 / 脳・神経 / 再生医学 / マイクロアレイ

Research Abstract

脊椎動物の初期胚は、受精後様々な細胞内シグナリングの作用による体軸形成・三胚葉分化を経て胚の基本的なボディパターンが規定される。これまで、Wnt、FGF、RA などのシグナリングの前後軸に沿った仮想的濃度勾配が脳や脊髄の部域を規定していることが示されてきたが、本課題では、特にどのような遺伝子の関与が脳、特に脳の部域既定に必要であるかを明らかにするため、以下の3点について解析を行った。(1)既存の脳マーカー遺伝子間の発現制御メカニズム解明を目指し、間脳特異的遺伝子Rx/Raxの、脳遺伝子Otx2と神経化因子Sox2による制御機構について研究を行った。その結果、Otx2蛋白とSox2蛋白は直接結合し、更にRx/Rax のプロモーター領域に結合して転写を制御することを見出した。(2)頭部で特異的に発現しかつWnt シグナルに制御を受ける新規遺伝子をマイクロアレイ解析によって約50同定した。その中で、ツメガエルの神経器官の一つであるセメント腺のみで特異的に発現する遺伝子としてXgalectinVIa、Xnkx3.1などを見出し、Wntシグナリングによる制御などを明らかにした。また、RA 代謝酵素XCyp26cがやはりWntシグナルに制御され、かつ一部のロンボメア特異的に発現することを見出した。更に、ヒストンメチルファラーゼをコードするPRDM12遺伝子が、口蓋プラコードで特異的に発現し、神経誘導能を有することを明らかにした。(3)予定神経外胚葉に存在する位置情報の根拠となる因子の同定を目指した。腹側割球由来の細胞と背側由来の細胞間で神経誘導能に差があることを見出した上で、これらの組織における遺伝子発現の差をマイクロアレイで解析した結果、いくつかの候補遺伝子を同定した。

Research Products

• [Journal Article] In vitro organogenesis fromundifferentiated cells in Xenopus (2009)
  • Author(s): Asashima M, Ito Y, Chan T, Michiue T,Nakanishi M, Suzuki K, Hitachi K, Okabayashi K, Kondow A, Ariizumi T
  • Journal Title: Dev Dyn in press
• [Journal Article] Decreased expression of CXXC4 promotesa malignant phenotype in renal cellcarcinoma by activating Wnt signaling (2009)
  • Author(s): Kojima T, Shimazui T, Hinotsu S, JorakuA, Oikawa T, Kawai K, Horie R, SuzukiH, Nagashima R, Yoshikawa K, Michiue T, Asashima M, Akaza H, Uchida K
  • Journal Title: Oncogene 28 Pages: 297-305
• [Journal Article] In vitro organogenesis from undifferentiated cells in Xenopus. (2009)
  • Author(s): Asashima et al.
  • Journal Title: Dev. Dyn. (印刷中(掲載確定))
  • Peer Reviewed
• [Journal Article] Retinoic acid metabolizingfactor xCyp26c is specificallyexpressed in neuroectoderm andregulates anterior neural patterningin Xenopus laevis (2008)
  • Author(s): Michiue T, Tanibe M, Yukita A, Danno H, Ikuzawa M, Ishiura S, Asashima M
  • Journal Title: Int. J. Dev. Biol 52 Pages: 893-901
• [Journal Article] nucleoredoxin regulates theWnt/planar cell polarity pathway inXenopus (2008)
  • Author(s): Funato Y, Michiue T, Terabayashi T,Yukita A, Danno H, Asashima M, Miki H
  • Journal Title: Gene Cells 13 Pages: 965-975
• [Journal Article] DevelopmentalPotential for Morphogenesis in Vivoand in Vitro (2008)
  • Author(s): Kaneko, K., Sato, K., Michiue, T., Okabayashi, K., Ohnuma, K., Danno, H., Asashima, M
  • Journal Title: J. Exp. Zool 310 Pages: 492-503
• [Journal Article] Wnt3a andDkk1 regulate distinctinternalization pathways of LRP6 totune the activation of β-cateninsignaling (2008)
  • Author(s): Yamamoto H, Sakane H, Yamamoto H, Michiue T, Kikuchi A
  • Journal Title: Dev. Cell 15 Pages: 37-48
• [Journal Article] Elucidation of the role ofactivin in organogenesis using amultiple organ induction system withamphibian and mouse undifferentiatedcells in vitro (2008)
  • Author(s): Asashima, M., Michiue, T., Kurisaki, A
  • Journal Title: Dev. Growth Differ.(Review) Supl1 Pages: 35-45
• [Journal Article] Molecular links among thecausative genes for ocularmalformation : Otx2 and Sox2 coregulateRax expression (2008)
  • Author(s): Danno, H., Michiue, T., Hitachi, K., Yukita, A., Ishiura, S., Asashima, M
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A 105 Pages: 5408-5413
• [Journal Article] Retinoic acid metabolizing factor xCyp26c is specifically expressed in neuroectoderm and regulates anterior neural patterning in Xenopus laevis. (2008)
  • Author(s): Tanibe et al.
  • Journal Title: Int. J. Dev. Biol. 52 Pages: 893-901
  • Peer Reviewed
• [Journal Article] Developmental Potential for Morphogenesis in Vivo and in Vitro. (2008)
  • Author(s): Kaneko et al.
  • Journal Title: J. Exp. Zool. 310 Pages: 492-503
  • Peer Reviewed
• [Journal Article] Molecular links among the causative genes for ocular malformation:Otx 2 and Sox 2 coregulate Rax expression (2008)
  • Author(s): Danno, et. al.
  • Journal Title: Proc.Nat.Acad.Sci.USA 105 Pages: 5408-5413
  • Peer Reviewed
• [Journal Article] Elucidation of the role of activin in organogenesis using a multiple organ induction system with amphibian and mouse undifferentiated cells in vitro (2008)
  • Author(s): Asashima, et. al.
  • Journal Title: Dev.Growth.Differ Supp.1
• [Journal Article] Xenopus galectin-VIa shows highlyspecific expression in cement glandsand is regulated by canonical Wntsignaling (2007)
  • Author(s): Michiue, T., Danno, H., Tanibe, M., Ikuzawa, M., Asashima, M
  • Journal Title: Gene. Expr. Patterns 7 Pages: 852-857
• [Journal Article] XSUMO-1 isrequired for normal mesoderm inductionand axis elongation during earlyXenopus development (2007)
  • Author(s): Yukita, A., Michiue, T., Danno, H., Asashima, M
  • Journal Title: Dev. Dyn 236 Pages: 2757-2766
• [Journal Article] Xenopus galectin-VIa shows highly specific expression in cement glands and is regulated by canonical Wnt signaling (2007)
  • Author(s): Michiue T., et. al.
  • Journal Title: Gen.EXPr.Patterns 7 Pages: 852-857
  • Peer Reviewed
• [Presentation] 眼形成不全原因遺伝子間の分子的関係 (2008)
  • Author(s): 團野宏樹, 道上達男, 石浦章一, 浅島誠
  • Organizer: 第1回XCIJ 首都圏支部会研究集会
  • Place of Presentation: 東京
  • Year and Date: 2008-10-18
• [Presentation] ヒトの眼の発生プログラム解明へのアプローチ (2008)
  • Author(s): 團野宏樹, 道上達男, 石浦章一, 浅島誠
  • Organizer: 東京大学生命科学研究ネットワークシンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2008-09-30
• [Presentation] Molecular relationship among theresponsible genes for ocularmalformation. (2008)
  • Author(s): Danno, H., Michiue, T., Hitachi, K.,Yukita, A., Ishiura, S., Asashima, M.
  • Organizer: The 41th Annual Meeting ofthe Japanese Society of DevelopmentalBiologists
  • Place of Presentation: 徳島
  • Year and Date: 2008-05-28
• [Presentation] Transcriptionalcontrol of eye and brain development : Otx2 and Sox2 coregulation. (2008)
  • Author(s): Danno, H., Michiue, T., Ishiura, S., Asashima, M
  • Organizer: The 12thInternational Xenopus Conference
  • Place of Presentation: Leiwen, Germany
• [Presentation] Exploration oftranscriptional networks controllinghead development by comparative genomicsapproach. (2008)
  • Author(s): Danno, H., Michiue, T., Ishiura, S., Asashima, M
  • Organizer: Frontiers in Developmental Biology
  • Place of Presentation: Hyeres, France
• [Presentation] Molecular links among the causative genes for ocular malformation: Otx2 and Sox2 coregulate Rax expression. (2008)
  • Author(s): Danno et al.
  • Organizer: 12th International Xenopus conference
  • Place of Presentation: Leiwen, Gernany