The regulation of tissue morphogenesis by non-coding RNAs

2012 Fiscal Year Final Research Report

• Project/Area Number: 22590266
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: UCHIJIMA Yasunobu 東京大学, 大学院・医学系研究科, 助手 (90272426)
• Project Period (FY): 2010 – 2012
• Keywords: noncoding RNA / miR-199a / miR-214 / 形態形成 / 骨形成 / 心筋傷害

Research Abstract

Our research group previously established the Dnm3os gene knockout mice. This gene encodes noncoding RNAs (ncRNAs) including miR-199a and miR-214. Since the mice displayed small body size with hypoplasia of bones and muscular tissues, the author assumed that these noncoding RNAs might regulate tissue morphogenesis. In order to clarify our hypothesis, the author knocked in miR-199a and/or miR-214 in the Dnm3os locus using the recombinase-mediated cassette exchange (RMCE) method. The authorconfirmed that the recovery of the expression of each miRNA among knocked in animals, but the recovery of abnormal phenotypes was observed only when both miRNAs were knocked in. Thus, it became clear that each noncoding RNA contributed to the tissue formation. The author tried to identify the target genes of these ncRNAs in P19 cells by using a unique dual drug selection system, but there seemed no gene displaying remarkable changes in expression. This could reflect the well-known feature of miR s that generally suppress the expression level of many genes with a small extent. As miR-199a and miR-214 were reported to change the expression levels in some heart problems, we suspected these genes could be involved in the pathogenesis of heart dysfunctions. Since our research group previously demonstrated that ETAR-positive cells contributed to the formation of heart and vessel systems, the author knocked in these ncRNAs to the ETAR locus using the RMCE method, and compared the histology of hearts with the control animals. Heart injury was induced by the administration of doxorubicin, which was known to cause severe heart injury in high dose in human. As a result, doxorubicin induced myocardial fibrosis in the control animals, but the fibrosis was considerably suppressed in the heart of miR-knocked in animals. These results suggested that miR-199a/214 might contribute to the maintenance of the normal heart function. Additionally, the results obtained here clearly demonstrated that this knock in system was an effective tool to investigate not only the mechanisms oftissue development but also the involvement of genes in disease.

Research Products

• [Journal Article] Preotic neural crest cells contribute to coronary artery smooth muscle involving endothelin signalling (2013)
  • Author(s): Arima Y, Miyagawa-Tomita S, Maeda K, Asai R, Seya D, Minoux m、Rijli FM, 西山 K, Kim KS, Uchijima Y, Ogawa H, Kurihara Y, Kurihara H.
  • Journal Title: Nature Communications Volume: 3 Pages: 1267-1277
  • Peer Reviewed
• [Journal Article] Angiogenic morphogenesis driven by dynamic and heterogeneous collective endothelial cell movement (2011)
  • Author(s): Arima S, Nishiyama K, Ko T, Arima Y, Hakozaki Y, Sugihara, Koseki H, Uchijima Y, Kurihara Y, Kurihara H.
  • Journal Title: Development Volume: 138 Pages: 4763-4776
  • Peer Reviewed
• [Journal Article] Identification and developmental analysis of endothelin receptor type-A expressiong cells in the mouse kidney (2011)
  • Author(s): Kitazawa T, Sato T, Nishiyama K, Asai R, Arima Y, Uchijima Y, Kurihara Y, Kurihara H.
  • Journal Title: Gene Expression Patterns Volume: 11 Pages: 371-377
  • Peer Reviewed
• [Journal Article] Calpain 6, a microtubule-stabilizing protein, regulates Rac1 activity and cell motility through interaction with GEF-H1 (2011)
  • Author(s): Tonami K, Kurihara Y, Arima S, Nishiyama K, Uchijima Y, Asano T, Sorimachi H, Kurihara H
  • Journal Title: Journal of Cell Sciences Volume: 124 Pages: 1214-1223
  • Peer Reviewed
• [Journal Article] Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest (2010)
  • Author(s): Kawamura Y, Uchijima Y, Horike N, Tonami K, Nishiyama K, Amano T, Asano T, Kurihara Y, Kurihara H.
  • Journal Title: Journal of Clinical Investigation Volume: 120 Pages: 2817-2828
  • Peer Reviewed
• [Journal Article] Endothelin receptor type-A expression defines a distinct cardiac subdomain within the heart field, with a later implication of this signaling pathway in chamber myocardium formation (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Miyagawa-Tomita S, Kurihara H.
  • Journal Title: Development Volume: 137 Pages: 3823-3833
  • Peer Reviewed
• [Journal Article] Establishment of mice expressiong EGFP in the placode-derived inner ear sensory cell lineage and FACS-array analysis focused on the regional specificity of the otocyst (2010)
  • Author(s): Fujimoto C, Ozeki H, Uchijima Y, Suzukawa K, Mitani A, Fukuhara S, nishiyama K, Kurihara Y, Kondo K, Aburatani H, Kaga K, Yamasoba T, Kurihara H.
  • Journal Title: Journal of Comparative Neurology Volume: 18 Pages: 4702-4722
  • Peer Reviewed
• [Presentation] Endothelin-1 regulates branchial arch morphogenesis by regulating intergenic enhanceractivity of Dlx5 and Dlx6 in mouse embryo (2012)
  • Author(s): Uchijima Y, Kurihara Y, Kushiyama S, Kurihara H.
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: マリンメッセ福岡(福岡県福岡市)
  • Year and Date: 2012-12-16
• [Presentation] MicroRNA 199/214 cluster is required for normal growth and skeletal development in mice (2012)
  • Author(s): Kawamura Y, Uchijima Y, Nishiyama K, Kurihara Y, Kita K, Kurihara H.
  • Organizer: Cell Symposia Functional RNAs
  • Place of Presentation: Hotel Melia (スペイン)
  • Year and Date: 2012-12-04
• [Presentation] Involvement of non-coding RNA: Evf2 in the Endothelin signaling in branchial arch development (2012)
  • Author(s): Kurihara, Y, Uchijima Y, Kushiyama S, Kurihara H
  • Organizer: Keystone Symposia Conference Noncoding RNAs
  • Place of Presentation: スノーバード・クリフロッジ(米国)
  • Year and Date: 2012-04-02
• [Presentation] novel roles of the neural crest in coronary artery formation through the Endothelin-1/Endothelin A receptor signaling (2012)
  • Author(s): Arima Y, Miyagawa-Tomita S, Nishiyama K, Asai R, Kim KS, Uchijima Y, Ogawa H, Kurihara Y. Kurihara H.
  • Organizer: 第76回日本循環器学会学術集会
  • Place of Presentation: 福岡国際会議場(福岡県福岡市)
  • Year and Date: 2012-03-18
• [Presentation] 下顎形成におけるエンドセリンシグナルにおけるnon-codingRNAとパラスペックル蛋白の作用 (2011)
  • Author(s): 栗原由紀子,内島泰信,櫛山櫻,濱崎真夏,西山功一,栗原裕基
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市)
  • Year and Date: 2011-12-13
• [Presentation] Coronary artery anomalies in Endothelin-1 and Endothelin A receptor knockout mice. (2011)
  • Author(s): Arima Y, Miyagawa-Tomita S, Nishiyama K, Asai R, Kim KS, Uchijima Y, Ogawa H, Kurihara Y, Kurihara H.
  • Organizer: American Heart Associatoin Scientific Session
  • Place of Presentation: オーラン ドコンベンションセンター(米国)
  • Year and Date: 2011-11-15
• [Presentation] Sirt3による参加ストレス防御機構の初期胚発生における役割 (2011)
  • Author(s): 河村悠美子,内島泰信,藤澤興,西山功一,栗原由紀子,栗原裕基
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都府京都市)
  • Year and Date: 2011-09-24
• [Presentation] Endothelin-1/Endothelin type-A receptor signaling regulates pharyngeal arch artery development through Dlx5/6-independent pathway. (2011)
  • Author(s): Kim KS, Arima Y, Asai R, Sato T, Uchijima Y, Nishiyama K, Kurihara Y, Igarashi T, Kurihara H.
  • Organizer: 211 Pediatric Academic Societies and Asian Society for Pediatric Research Jont Meeting
  • Place of Presentation: Denver Convention Center(米国)
  • Year and Date: 2011-05-01
• [Presentation] Endothelin type-A receptor expression defines a distinct subpopulation within the first heart field contributing to chamber myocardium (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Saga Y, Miyagawa-Tomita S, Kurihara H.
  • Organizer: 第33回日本分子生物学会・第 83 回日本 生化学会 合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県神戸市)
  • Year and Date: 2010-12-10
• [Presentation] Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest (2010)
  • Author(s): Kawamura Y, Uchijima Y, Horike N, Tonami K, Nishiyama K, Asano T, Amano T, Kurihara Y, Kurihara H.
  • Organizer: 第33回日本分子生物学会・第 83 回日本生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県神戸市)
  • Year and Date: 2010-12-09
• [Presentation] Transcriptional regulation of non-coding RNA Evf-2 in the Endothelin-1/Dlx5/Dlx6 pathway (2010)
  • Author(s): Uchijima Y, Kurihara Y, Sato T, Fujisawa K, Kushiyama S, Kurihara H.
  • Organizer: 第33回 日本分子生物学会・第 83 回 日本 生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県神戸市)
  • Year and Date: 2010-12-08
• [Remarks]
  • URL: http://bio.m.u-tokyo.ac.jp/