Development of locus- and/or neurotransmitter-specific cell depletion method in CNS and/or sensory organs

• Project/Area Number: 16H04688
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: YONEKAWA Hiromichi 公益財団法人東京都医学総合研究所, 基盤技術研究センター, 特任研究員 (30142110)
• Co-Investigator(Kenkyū-buntansha): 吉川 欣亮 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, プロジェクトリーダー (20280787) ; 設楽 浩志 公益財団法人東京都医学総合研究所, 基盤技術研究センター, 主席基盤技術研究職員 (90321885)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2018: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2016: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
• Keywords: 毒素受容体 / ジフテリア毒素 / 組織特異的プロモーター / トランスジェニックマウス / 蛍光タンパク質 / Cre/loxPシステム / ヒト疾患モデルマウス / ノックインマウス / ジフテリア毒素受容体 / 遺伝子改変マウス / 中枢神経系 / 蛍光蛋白 / 組織特異的プロモータ / Cre/loxP系 / Dre/rox系 / 蛍光蛋白質

Outline of Final Research Achievements

TRECK enable us to deplete tissue-specifically any cells of interest in live mice and the depletion lead us to disclose in vivo function of the cell. We have succeeded in generate TRECK-Tg mice, the origin of the cells are endoderm or mesoderm. However, we did not succeed in generate TRECK-Tg mice for ectodermal cells. Therefore, we tried to develop CNS-, or neurotransmitter- specific TRECK method, with which we also combined color-differential display of brain by fluorescent proteins. At the first, although we used the genes which play an important roles in early development of CNS, the expression of fluorescent proteins was too weak to radicalize for conventional fluorescent microscopy. Next, we used the gene which strongly expresses in adult brain, and found that the fluorescent protein is strongly expressed enough to do the macro-scale observation in fluorescent microscopy. Now, we are investigating whether color display and specific cell depletion take place.

Academic Significance and Societal Importance of the Research Achievements

我々はこれまで、内胚葉や中胚葉起源の細胞をマウス体内から特異的に除去する方法(TRECK法)を開発し、ヒト疾患モデルマウスの樹立や、その標的となる細胞の個体内での機能を明らかにしてきた。今回、我々は未着手の中枢神経系に対し、この方法の応用を試みた。また、この方法には脳内を部署ごとに蛍光蛋白で識別化が可能なようなDNA組換え体を考案した。様々な試行錯誤の結果、蛍光顕微鏡下で識別化が可能となるような有望なマウスを作出した。このマウスは脳の蛍光による識別化と、最終的には脳内から1種類の細胞を除去できることが期待される。従って、この研究の成果が成就したときは、脳科学の方面に大きな貢献が期待できる。

Research Products

• [Journal Article] OHC-TRECK: A Novel System Using a Mouse Model for Investigation of the Molecular Mechanisms Associated with Outer Hair Cell Death in the Inner Ear (2019)
  • Author(s): Matsuoka Kunie、Wada Kenta、Miyasaka Yuki、Yasuda Shumpei P.、Seki Yuta、Nishito Yasumasa、Yonekawa Hiromichi、Taya Choji、Shitara Hiroshi、Kikkawa Yoshiaki
  • Journal Title: Scientific Reports Volume: 9: 5285 Issue: 1
  • DOI: 10.1038/s41598-019-41711-2
  • Peer Reviewed / Open Access
• [Journal Article] Mice deficient in the Shmt2 gene have mitochondrial respiration defects and are embryonic lethal. (2018)
  • Author(s): Tani H, Ohnishi S, Shitara H, Mito T, Yamaguchi M, Yonekawa H, Hashizume O, Ishikawa K, Nakada K, Hayashi JI.
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 425-425
  • DOI: 10.1038/s41598-017-18828-3
  • NAID: 120007134501
  • Peer Reviewed / Open Access
• [Journal Article] Pde6b rd1 mutation modifies cataractogenesis in Foxe3 rct mice (2018)
  • Author(s): Wada Kenta、Saito Junichi、Yamaguchi Midori、Seki Yuta、Furugori Masamune、Takahashi Gou、Nishito Yasumasa、Matsuda Hiroshi、Shitara Hiroshi、Kikkawa Yoshiaki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 496 Issue: 1 Pages: 231-237
  • DOI: 10.1016/j.bbrc.2018.01.031
  • Peer Reviewed
• [Journal Article] Establishment of a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes and improvement of a PCR-RFLP-based measurement system for estimation of mitochondrial DNA heteroplasmy (2017)
  • Author(s): H. Shitara, L. Cao, M. Yamaguchi, H. Yonekawa、C. Taya
  • Journal Title: Transgenic Res. Volume: 26 Issue: 4 Pages: 559-565
  • DOI: 10.1007/s11248-017-0009-2
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Skin CD4+ Memory T Cells Play an Essential Role in Acquired Anti-Tick Immunity through Interleukin-3-Mediated Basophil Recruitment to Tick-Feeding Sites (2017)
  • Author(s): Ohta Takuya、Yoshikawa Soichiro、Tabakawa Yuya、Yamaji Kayoko、Ishiwata Kenji、Shitara Hiroshi、Taya Choji、Oh-hora Masatsugu、Kawano Yohei、Miyake Kensuke、Yamanishi Yoshinori、Yonekawa Hiromichi、Watanabe Naohiro、Kanuka Hirotaka、Karasuyama Hajime
  • Journal Title: Frontiers in Immunology Volume: 8 Pages: 1348-1348
  • DOI: 10.3389/fimmu.2017.01348
  • Peer Reviewed / Open Access
• [Journal Article] A novel splice site mutation of myosin VI in mice leads to stereociliary fusion caused by disruption of actin networks in the apical region of inner ear hair cells (2017)
  • Author(s): Seki Yuta、Miyasaka Yuki、Suzuki Sari、Wada Kenta、Yasuda Shumpei P.、Matsuoka Kunie、Ohshiba Yasuhiro、Endo Kentaro、Ishii Rie、Shitara Hiroshi、Kitajiri Shin-ichiro、Nakagata Naomi、Takebayashi Hirohide、Kikkawa Yoshiaki
  • Journal Title: PLOS ONE Volume: 12 Issue: 8 Pages: e0183477-e0183477
  • DOI: 10.1371/journal.pone.0183477
  • NAID: 120007099799
  • Peer Reviewed / Open Access
• [Journal Article] A novel missense mutation of <i>Mip</i> causes semi-dominant cataracts in the <i>Nat</i> mouse (2017)
  • Author(s): Takahashi Gou、Hasegawa Sayaka、Fukutomi Yukiko、Harada Chihiro、Furugori Masamune、Seki Yuta、Kikkawa Yoshiaki、Wada Kenta
  • Journal Title: Experimental Animals Volume: 66 Issue: 3 Pages: 271-282
  • DOI: 10.1538/expanim.17-0012
  • NAID: 130005907568
  • ISSN: 0007-5124, 1341-1357, 1881-7122
  • Peer Reviewed
• [Journal Article] The parathyroid hormone regulates skin tumour susceptibility in mice (2017)
  • Author(s): Okumura Kazuhiro、Saito Megumi、Yoshizawa Yasuhiro、Munakata Haruka、Isogai Eriko、Miura Ikuo、Wakana Shigeharu、Yamaguchi Midori、Shitara Hiroshi、Taya Choji、Karaplis Andrew C.、Kominami Ryo、Wakabayashi Yuichi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 11208-11208
  • DOI: 10.1038/s41598-017-11561-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Heterozygous mutation of Ush1g/Sans in mice causes early-onset progressive hearing loss, which is recovered by reconstituting the strain-specific mutation in Cdh23 (2016)
  • Author(s): Miyasaka Y, Shitara H, Suzuki S, Yoshimoto S, Seki Y, Ohshiba Y, Okumura K, Taya C, Tokano H, Kitamura K, Takada T, Hibino H, Shiroishi T, Kominami R, Yonekawa H, Kikkawa Y
  • Journal Title: Hum. Mol. Genet. Volume: 印刷中 Issue: 10 Pages: 2045-2059
  • DOI: 10.1093/hmg/ddw078
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Essential contribution of CD4+ T cells to antigen-induced nasal hyperresponsiveness in experimental allergic rhinitis. (2016)
  • Author(s): Nishimura, T., Kaminuma, O., Saeki, M., Kitamura, N., Matsuoka, K., Yoneyama, H., Mori, A., and Hiroi, T.
  • Journal Title: Plos One. Volume: １１ Issue: 1 Pages: e0146686-e0146686
  • DOI: 10.1371/journal.pone.0146686
  • Peer Reviewed / Open Access