Overexpression of myogenic regulatory factors driven by heat shock promoters in medaka and its developmental study.

• Project/Area Number: 22700428
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Neurophysiology and muscle physiology
• Research Institution: Keio University
• Principal Investigator: SHIMIZU Atsushi 慶應義塾大学, 医学部, 助教 (30327655)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 受容体 / 細胞内シグナル伝達 / 再生分化 / ACVR1 / ALK2 / FOP / 熱ショックプロモーター / メダカ / 骨異形成症 / モデル生物 / ゼブラフィッシュ / トランスジェニック / Cre loxP / 熱ショックタンパク

Research Abstract

Fibrodysplasia ossificans progressiva(FOP) is a genetic disorder of dysregulated cellular differentiation which causes fibrous tissue(including muscle, tendon, and ligament) to be ossified when damaged. The mutations were identified in the activin A receptor, type I(ACVR1), a BMP type-1receptor. This study was purposed to establish the transgenic medaka fish that can induce overexpression of the ACVR1or BMP4gene by heat-shock in any target time points, tissues and cells. I established heat-shock inducible Cre recombinase expressing transgenic medaka fish(hsp70b :: Cre) using endogenous hsp70promoter and Cre inducible myogenic cells specific ACVR1expressing transgenic fish(DES :: lGlBMP4). However, both of heat-shock treated and non-treated double transgenic medaka fishes(hsp70b :: Cre and DES :: lGlBMP4) exhibited FOP like phenotypes, which mean endogenous hsp70 promoter was expressed continuously at a low level without heat induction. Then I combined the dual promoter expressing system and artificial heat-shock promoter and established heat-inducible double transgenic medaka fish(8xHSE :: Cre x DES : lGlACVR1).

Research Products

• [Journal Article] The Medaka zic1/zic4Mutant Provides Molecular Insights into Teleost Caudal Fin Evolution (2012)
  • Author(s): Moriyama Y, Kawanishi T, Nakamura R, Tsukahara T, Sumiyama K, Suster ML, Kawakami K, Toyoda A, Fujiyama A, Yasuoka Y, Nagao Y, Sawatari E, Shimizu A, Wakamatsu Y, Hibi M, Taira M, Okabe M, Naruse K, Hashimoto H, Shimada A and Takeda H
  • Journal Title: Curr Biol Volume: 22 Pages: 601-607
  • Peer Reviewed
• [Journal Article] The medaka zic1/zic4 mutant provides molecular insights into teleost caudal fin evolution. (2012)
  • Author(s): Yuuta Moriyama
  • Journal Title: Current Biology Volume: 21 Issue: 7 Pages: 601-607
  • DOI: 10.1016/j.cub.2012.01.063
  • Peer Reviewed
• [Journal Article] Insufficiency of BUBR1, a mitotic spindle checkpoint regulator, causes impaired ciliogenesis in vertebrates (2011)
  • Author(s): Miyamoto T, Porazinski S, Wang H, Borovina A, Ciruna B, Shimizu A, Kajii T, Kikuchi A, Furutani-Seiki M, and Matsuura S
  • Journal Title: Human Molecular Genetics Volume: 20 Pages: 2058-2070
  • Peer Reviewed
• [Journal Article] 次世代シークエンサーによるトランスクリプトーム解析 (2011)
  • Author(s): 清水厚志
  • Journal Title: 臨床検査 Volume: 55 Pages: 841-846
• [Journal Article] Insufficiency of BUBR1, a mitotic spindle checkpoint regulator, causes impaired ciliogenesis in vertebrates. (2011)
  • Author(s): Miyamoto T, Shimizu A, et al
  • Journal Title: Human Molecular Genetics Volume: (in press)
  • Peer Reviewed
• [Journal Article] YPEL5protein of the YPEL gene family is involved in the cell cycle progression by interacting with two distinct proteins RanBPM and RanBP10 (2010)
  • Author(s): Hosono K, Noda S, Shimizu A, Nakanishi N, Ohtsubo M, Shimizu N and Minoshima S
  • Journal Title: Genomics Volume: 96 Pages: 102-111
  • Peer Reviewed
• [Journal Article] メダカを用いたヒト疾患モデル (2010)
  • Author(s): 清水厚志、亀井保博、清水信義
  • Journal Title: 比較内分泌学 Volume: 36(139) Pages: 286-292
  • NAID: 10027129728
• [Journal Article] TILLING法による遺伝子変異メダカの作製 (2010)
  • Author(s): 貴田亨、石川智子、清水厚志、亀井保博、殿山泰弘、藤堂剛、三輪正直、清水信義
  • Journal Title: 比較内分泌学 Volume: 36(137) Pages: 154-158
• [Journal Article] メダカ変異体を使った個体レベルのゲノム研究の展望 (2010)
  • Author(s): 亀井保博、清水厚志、清水信義
  • Journal Title: 比較内分泌学 Volume: 36(136) Pages: 66-68
  • NAID: 10025990367
• [Journal Article] YPEL5 protein of the YPEL gene family is involved in the cell cycle progression by interacting with two distinct proteins RanBPM and RanBP10. (2010)
  • Author(s): Hosono K, Shimizu A, et al
  • Journal Title: Genomics Volume: 96 Pages: 102-111
  • Peer Reviewed
• [Presentation] Medaka Model for Fibrodysplasia Ossificans Progressiva (2010)
  • Author(s): Shimizu, A. and Shimizu N
  • Organizer: The American Society of Human Genetics60th Annual Meeting
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2010-11-03
• [Presentation] Medaka Model for Fibrodysplasia Ossificans Progressiva. (2010)
  • Author(s): Shimizu, A., Shimizu, N
  • Organizer: The American Society of Human Genetics 60th Annual Meeting
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2010-11-03
• [Remarks] 作製したトランスジェニックラインは「NBRPメダカ」に順次寄託している。
  • URL: http://www.shigen.nig.ac.jp/medaka/