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In the present study, we focused on the role of reactive oxygen species (ROS) produced by the NADPH oxidase Dual oxidase (Duox) during cricket leg regeneration. The results showed that ROS levels were upregulated during leg regeneration and decreased by DuoxRNAi. In DuoxRNAi nymphs, wound closure and scab formation were incomplete 2\u2005days after amputation, and hypertrophy occurred in the distal region of the regenerating legs at 5\u2005days after amputation. In addition, the hypertrophic phenotype was induced by DuoxARNAi and NADPH oxidase inhibitor treatment. During hypertrophy, haemocytes, including plasmatocytes, oenocytoids and granulocytes, accumulated. Proliferation of haemocytes in regenerating legs was not increased by DuoxRNAi; however, haemocyte accumulation was regulated by the Spatzle (Spz) family molecules, which are Toll receptor ligands. As the exoskeleton of DuoxRNAi nymphs was thinner than that of the control, excessive haemocyte accumulation can cause hypertrophy in DuoxRNAi nymphs. Thus, Duox-derived ROS are involved in wound healing and haemocyte accumulation through the Spz\/Toll signalling pathway during leg regeneration in crickets.<\/jats:p>","DOI":"10.1242\/dev.204763","type":"journal-article","created":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T09:56:42Z","timestamp":1763632602000},"source":"Crossref","is-referenced-by-count":1,"title":["ROS produced by Dual oxidase regulate cell proliferation and haemocyte migration during leg regeneration in the cricket"],"prefix":"10.1242","volume":"152","author":[{"given":"Misa","family":"Okumura-Hirono","sequence":"first","affiliation":[{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 1 Department of Cytology and Histology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]},{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 2 Department of Urology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3038-7782","authenticated-orcid":false,"given":"Tetsuya","family":"Bando","sequence":"additional","affiliation":[{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 1 Department of Cytology and Histology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7143-5365","authenticated-orcid":false,"given":"Yoshimasa","family":"Hamada","sequence":"additional","affiliation":[{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 1 Department of Cytology and Histology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]},{"name":"Institute for Genome Research, Institute of Advanced Medical Sciences, Tokushima University 3 Division of Molecular Biology , , 3-18-15, Kuramoto-cho, Tokushima 770-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3172-2482","authenticated-orcid":false,"given":"Motoo","family":"Araki","sequence":"additional","affiliation":[{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 2 Department of Urology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1961-606X","authenticated-orcid":false,"given":"Hideyo","family":"Ohuchi","sequence":"additional","affiliation":[{"name":"Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 1 Department of Cytology and Histology , , 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8558 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2025,11,24]]},"reference":[{"key":"2026090107090310900_DEV204763C1","doi-asserted-by":"publisher","first-page":"e1005746","DOI":"10.1371\/journal.ppat.1005746","article-title":"Transdifferentiation and proliferation in two distinct hemocyte lineages in Drosophila melanogaster larvae after wasp infection","volume":"12","author":"Anderl","year":"2016","journal-title":"PLoS Pathog."},{"key":"2026090107090310900_DEV204763C2","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1093\/genetics\/163.1.203","article-title":"Analysis of Ras-induced overproliferation in Drosophila hemocytes","volume":"163","author":"Asha","year":"2003","journal-title":"Genetics"},{"key":"2026090107090310900_DEV204763C3","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1002\/dvdy.5","article-title":"Amputation-induced reactive oxygen species signaling is required for Axolotl tail regeneration","volume":"248","author":"Baddar","year":"2019","journal-title":"Dev. 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