{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T16:25:05Z","timestamp":1771259105135,"version":"3.50.1"},"reference-count":33,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T00:00:00Z","timestamp":1676505600000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21K06195"],"award-info":[{"award-number":["21K06195"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Dev Growth Differ"],"published-print":{"date-parts":[[2023,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Amphibians and fish often regenerate lost parts of their appendages (tail, limb, and fin) after amputation. Limb regeneration in adult amphibians provides an excellent model for appendage (limb) regeneration through 3D morphogenesis along the proximodistal, dorsoventral, and anteroposterior axes in mammals, because the limb is a homologous organ among amphibians and mammals. However, manipulating gene expression in specific appendages of adult amphibians remains difficult; this in turn hinders elucidation of the molecular mechanisms underlying appendage regeneration. To address this problem, we devised a system for appendage\u2010specific gene induction using a simplified protocol named the \u201cagarose\u2010embedded heat shock (AeHS) method\u201d involving the combination of a heat\u2010shock\u2010inducible system and insertion of an appendage in a temperature\u2010controlled agarose gel. Gene expression was then induced specifically and ubiquitously in the regenerating limbs of metamorphosed amphibians, including a frog (<jats:italic>Xenopus laevis<\/jats:italic>) and newt (<jats:italic>Pleurodeles waltl<\/jats:italic>). We also induced gene expression in the regenerating tail of a metamorphosed <jats:italic>P. waltl<\/jats:italic> newt using the same method. This method can be applied to adult amphibians with large body sizes. Furthermore, this method enables simultaneous induction of gene expression in multiple individuals; further, the data are obtained in a reproducible manner, enabling the analysis of gene functions in limb and tail regeneration. Therefore, this method will facilitate elucidation of the molecular mechanisms underlying appendage regeneration in amphibians, which can support the development of regenerative therapies for organs, such as the limbs and spinal cord.<\/jats:p>","DOI":"10.1111\/dgd.12841","type":"journal-article","created":{"date-parts":[[2023,1,21]],"date-time":"2023-01-21T14:30:32Z","timestamp":1674311432000},"page":"86-93","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Appendage\u2010restricted gene induction using a heated agarose gel for studying regeneration in metamorphosed <i>Xenopus laevis<\/i> and <i>Pleurodeles waltl<\/i>"],"prefix":"10.1111","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3974-6630","authenticated-orcid":false,"given":"Haruka","family":"Matsubara","sequence":"first","affiliation":[{"name":"School of Life Science, Faculty of Medicine Tottori University  Tottori Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8118-8548","authenticated-orcid":false,"given":"Aiko","family":"Kawasumi\u2010Kita","sequence":"additional","affiliation":[{"name":"Laboratory for Developmental Morphogeometry RIKEN Center for Biosystems Dynamics Research  Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2565-8944","authenticated-orcid":false,"given":"Saki","family":"Nara","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science Hirosaki University  Aomori Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1414-8802","authenticated-orcid":false,"given":"Hibiki","family":"Yokoyama","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science Hirosaki University  Aomori Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3676-5710","authenticated-orcid":false,"given":"Toshinori","family":"Hayashi","sequence":"additional","affiliation":[{"name":"Amphibian Research Center \/ Graduate School of Integrated Sciences for Life Hiroshima University  Hiroshima Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takashi","family":"Takeuchi","sequence":"additional","affiliation":[{"name":"School of Life Science, Faculty of Medicine Tottori University  Tottori Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4821-9506","authenticated-orcid":false,"given":"Hitoshi","family":"Yokoyama","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science Hirosaki University  Aomori 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