Local gene induction system in an amphibian and its application to morphogenetic regeneration

• Project/Area Number: 25870058
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology; Morphology/Structure
• Research Institution: Hirosaki University (2014-2016); Tohoku University (2013)
• Principal Investigator: Yokoyama Hitoshi 弘前大学, 農学生命科学部, 准教授 (90455816)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2014: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 再生 / 両生類 / 遺伝子発現 / 四肢 / 器官形成 / 形態形成

Outline of Final Research Achievements

Amphibians have high regenerative capacity. For example, they can regenerate a limb after amputation. However, difficulty in manipulating gene expression in spatially restricted manner has so far hindered elucidation of the molecular mechanisms of regeneration in amphibians. In this study, I applied two types of gene induction systems, IR-LEGO and temperature stimulator to amphibians and enabled local gene induction. As a result, function of a specific gene has been revealed by single cell level gene induction by IR-LEGO. Furthermore, gene induction exclusively in a specific region in a limb has been done by using temperature stimulator.

Research Products

• [Journal Article] Developmental stages for the divergence of relative limb length between a twig and a trunk-ground Anolis lizard species (2015)
  • Author(s): Wakasa, H., Cadiz, A., Echenique-Diaz, L.M. Iwasaki, W.M. Kamiyama, N., Nishimura, Y., Yokoyama, H., Tamura, K. and Kawata, M.
  • Journal Title: Journal of Experimental Zoology Part B: Molecular and Developmental Evolution Volume: 未定 Issue: 5 Pages: 410-423
  • DOI: 10.1002/jez.b.22627
  • Peer Reviewed
• [Journal Article] Evidence for an amphibian sixth digit. (2015)
  • Author(s): Hayashi S., Kobayashi T., Yano T., Kamiyama N., Egawa S., Seki R., Takizawa K., Okabe M., Yokoyama H. and Tamura K.
  • Journal Title: Zoological Letters. Volume: 1 Issue: 1 Pages: 17-17
  • DOI: 10.1186/s40851-015-0019-y
  • Peer Reviewed / Open Access
• [Journal Article] Epigenetic modification maintains intrinsic limb-cell identity in Xenopus limb bud regeneration. (2015)
  • Author(s): Hayashi, S., Kawaguchi, A., Uchiyama, I., Kawasumi-Kita, A., Kobayashi, T., Nishide, H., Tsutsumi, R., Tsuru, K., Inoue, T., Ogino, H., Agata, K., Tamura, K. and *Yokoyama, H.
  • Journal Title: Dev. Biol. Volume: 406 Issue: 2 Pages: 271-282
  • DOI: 10.1016/j.ydbio.2015.08.013
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] AP-2β is a transcriptional regulator for determination of digit length in tetrapods. (2015)
  • Author(s): Seki R, Kitajima K, Matsubara H, Suzuki T, Saito D, Yokoyama H, Tamura K.
  • Journal Title: Dev Biol. Volume: 407 Issue: 1 Pages: 75-89
  • DOI: 10.1016/j.ydbio.2015.08.006
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Application of local gene induction by infrared laser-mediated microscope and temperature stimulator to amphibian regeneration study. (2015)
  • Author(s): Kawasumi, A., Hayashi, T., Kobayashi, T., Nagayama, C., Hayashi, S., Kamei, Y., Morishita, Y., T Takeuchi, T., Tamura, K., Yokoyama, H.
  • Journal Title: Dev. Growth Differ. Volume: 57 Issue: 9 Pages: 601-613
  • DOI: 10.1111/dgd.12241
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Roles of Hippo signaling pathway in size control of organ regeneration (2015)
  • Author(s): Hayashi, S., Yokoyama, H. and Tamura, K.
  • Journal Title: Development Growth and Differentiation Volume: 未定 Issue: 4 Pages: 341-351
  • DOI: 10.1111/dgd.12212
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Altered developmental events in the anterior region of the chick forelimb give rise to avian-specific digit loss (2014)
  • Author(s): Naoki Nomura, Hitoshi Yokoyama and Koji Tamura
  • Journal Title: Developmental Dynamics Volume: published online Issue: 6 Pages: 741-52
  • DOI: 10.1002/dvdy.24117
  • Peer Reviewed / Open Access
• [Journal Article] Growth and differentiation of a long bone in limb development, repair and regeneration (2014)
  • Author(s): S. Egawa, S. Miura, H. Yokoyama, T. Endo and K. Tamura
  • Journal Title: Dev Growth Differ Volume: 56 Issue: 5 Pages: 410-24
  • DOI: 10.1111/dgd.12136
  • NAID: 40020109556
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Transcriptional regulators in the Hippo signaling pathway control organ growth in Xenopus tadpole tail regeneration (2014)
  • Author(s): S. Hayashi, H. Ochi, H. Ogino, A. Kawasumi, Y. Kamei, K. Tamura and H. Yokoyama
  • Journal Title: Dev Biol Volume: 396 Issue: 1 Pages: 31-41
  • DOI: 10.1016/j.ydbio.2014.09.018
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] アフリカツメガエルから見た四肢再生実現へのステップ (2014)
  • Author(s): 横山仁、林真一、川住愛子、砂川奈都召、田村宏治
  • Journal Title: 実験医学 Volume: 32(1) Pages: 22-28
• [Journal Article] Yap1, transcription regulator in the Hippo signaling pathway, is required for Xenopus limb bud regeneration. (2014)
  • Author(s): Hayashi S, Tamura K, Yokoyama H
  • Journal Title: Developmental Biology Volume: 388 Issue: 1 Pages: 57-67
  • DOI: 10.1016/j.ydbio.2014.01.018
  • Peer Reviewed / Open Access
• [Journal Article] Growth and differentiation of a long bone in limb development, repair and regeneration (2014)
  • Author(s): Egawa, S., Miura, S., Yokoyama, H., Endo, T., and Tamura, K.
  • Journal Title: Development Growth and Differentiation Volume: 未定
  • NAID: 40020109556
  • Peer Reviewed
• [Journal Article] 両生類の皮膚再生と四肢再生の関係から見る，両生類再生研究の哺乳類への応用の可能性 (2013)
  • Author(s): 田村宏治、大塚理奈、川住愛子、横山仁
  • Journal Title: 瘢痕・ケロイド治療ジャーナル Volume: ７ Pages: 1-4
• [Journal Article] 四肢再生における脱分化、再分化と細胞記憶 (2013)
  • Author(s): 林真一、矢野十織、川住愛子、田村宏治、横山仁
  • Journal Title: 実験医学 Volume: 31(13) Pages: 2075-2082
• [Presentation] 四肢と皮膚の完全再生を可能にする分子・細胞メカニズムの探求 (2016)
  • Author(s): 横山仁
  • Organizer: 日本解剖学会 シンポジウム再生研究の新機軸 ―不可能を可能とする内在性プログラムの探索―
  • Place of Presentation: ビッグパレットふくしま（福島県郡山市）
  • Year and Date: 2016-03-28
  • Invited
• [Presentation] Skin regeneration of Xenopus, as a model for scar-free skin regeneration in adulthood (2016)
  • Author(s): Hitoshi Yokoyama, Rina Otsuka, Aiko Kawasumi-Kita, Tamae Maruoka and Koji Tamura
  • Organizer: Aquatic model organisms for human disease and toxicology research
  • Place of Presentation: 岡崎コンファレンスセンター（愛知県岡崎市）
  • Year and Date: 2016-03-18
  • Int'l Joint Research / Invited
• [Presentation] ツメガエルにおける局所的な遺伝子発現の操作法の確立と再生研究への応用 (2015)
  • Author(s): 横山仁, 川住愛子, 林利憲, 小林託也, 永山誓史, 林真一, 亀井保博, 森下喜弘, 竹内隆, 田村宏治
  • Organizer: 日本ツメガエル研究集会
  • Place of Presentation: 秋田温泉さとみ（秋田市）
  • Year and Date: 2015-09-15
• [Presentation] アフリカツメガエルを用いた皮膚再生の研究 ‐これまでとこれから‐ (2015)
  • Author(s): 横山仁
  • Organizer: 次世代両生類研究会
  • Place of Presentation: 岡崎コンファレンスセンター（愛知県岡崎市）
  • Year and Date: 2015-08-24
  • Invited
• [Presentation] アフリカツメガエルにおける皮膚の完全再生に寄与する細胞の起源 (2015)
  • Author(s): 大塚理奈, 有泉高史, 永山誓史, 川住愛子, 田村宏治, 横山仁
  • Organizer: 日本動物学会東北支部大会
  • Place of Presentation: 東北大学農学部（仙台市）
  • Year and Date: 2015-08-09
• [Presentation] アフリカツメガエルにおける瘢痕を残さない皮膚再生 ‐再生に寄与する細胞はどこから来るのか？- (2015)
  • Author(s): 横山仁、大塚理奈、有泉高史、永山誓史、川住愛子、田村宏治
  • Organizer: 日本再生医療学会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2015-03-21
• [Presentation] Cellular origin of blastema-like cells contributing to scarless skin regeneration of metamorphosed Xenopus laevis (2014)
  • Author(s): Rina Otsuka, Chikashi Nagayama, Aiko Kawasumi, Takashi Ariizumi , Koji Tamura, Hitoshi Yokoyama
  • Organizer: 日本発生生物学会
  • Place of Presentation: ウインクあいち（名古屋市）
  • Year and Date: 2014-05-28 – 2014-05-29
• [Presentation] Cellular origin of blastema-like cells contributing to scarless (2014)
  • Author(s): Rina Otsuka, Chikashi Nagayama, Aiko Kawasumi, Takashi Ariizumi, Koji Tamura, Hitoshi Yokoyama
  • Organizer: 日本発生生物学会
  • Place of Presentation: ウィンクあいち
• [Presentation] Different cellular contribution of somite-derived tissues to limb regeneration in larval and adult Xenopus (2013)
  • Author(s): 永山誓史、有泉高史 、田村宏治、横山仁
  • Organizer: 日本発生生物学会
  • Place of Presentation: くにびきメッセ（島根県松江市）
• [Presentation] 発生原理から見たアフリカツメガエルの器官再生 (2013)
  • Author(s): 横山仁
  • Organizer: 日本動物学会
  • Place of Presentation: 岡山大学
  • Invited
• [Remarks] 弘前大学農学生命科学部教員紹介ページ
  • URL: http://nature.cc.hirosaki-u.ac.jp/staff/hitoshi-yokoyama
• [Remarks] 弘前大学農学生命科学部教員紹介
  • URL: http://nature.cc.hirosaki-u.ac.jp/public/hitoshi-yokoyama/
• [Remarks] 田村研ホームページ 横山仁 研究業績リスト
  • URL: http://www.biology.tohoku.ac.jp/lab-www/tamlab/members/yoko_pub.html
• [Remarks] 東北大学 大学院生命科学研究科 教員紹介ページ 横山仁
  • URL: http://www.lifesci.tohoku.ac.jp/teacher/t_yokoyamaj/