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J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Anti-M\u00fcllerian hormone (Amh) is a key regulator of sex determination and differentiation in fish. In this study, we identified and characterized a single-copy Amh ortholog, designated CmAmh, in blotched snakehead (Channa maculata), an economically important species in China. The cDNA spans 2340 bp and encodes a 545-amino-acid protein with conserved AMH-N and TGF-\u03b2 domains. CmAmh exhibited pronounced male-biased gonadal expression, with transcript levels in testes markedly exceeding those in ovaries across all developmental stages (45~365 days post-hatching (dph)). Cellular localization revealed CmAmh transcripts in the Sertoli cells and spermatogonia of the testis and, more weakly, in the oogonia and primary oocytes of the ovary, suggesting a primary role in spermatogenesis and a potential modulatory function in early oogenesis. A strong inverse correlation between promoter CpG methylation and transcriptional activity, hypermethylation in ovaries (82.5~89.7%) versus hypomethylation (15.8~19.1%) in testes, indicated epigenetic regulation, which functionally integrates with hormonal signaling: 17\u03b2-estradiol (E2) suppressed, whereas methyltestosterone (MT) upregulated CmAmh expression. Long-term E2 exposure induced male-to-female secondary sex reversal (SSR) in 26.7% of XY individuals, whereas MT failed to reverse XX fish, indicating that CmAmh modulation alone is insufficient to override ovarian fate. Furthermore, CRISPR\/Cas9-mediated mutagenesis achieved 40~45% editing efficiency, predominantly generating frameshift alleles, providing a platform for future functional studies. Collectively, these findings position CmAmh as a hormone-responsive nodal point within a broader sex-differentiation network. This work provides a theoretical foundation for sex-control strategies in C. maculata aquaculture, while future studies should focus on homozygous mutant generation to definitively establish its loss-of-function phenotype and regulatory networks.<\/jats:p>","DOI":"10.3390\/ijms27198588","type":"journal-article","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T10:34:10Z","timestamp":1790332450000},"page":"8588","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Role of Amh in Gonadal Development and Its Response to Exogenous Hormones in Blotched Snakehead (Channa maculata)"],"prefix":"10.3390","volume":"27","author":[{"given":"Junqi","family":"Qin","sequence":"first","affiliation":[{"name":"China (Guangxi)-ASEAN Key Laboratory of Comprehensive Exploitation and Utilization of Aquatic Germplasm Resources, Ministry of Agriculture and Rural Affairs, Key Laboratory of Aquaculture Genetic and Breeding and Healthy Aquaculture of Guangxi, Guangxi Academy of Fishery Sciences, Nanning 530021, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Yueying","family":"Deng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical and Subtropical Fishery Resources Application and Cultivation, Ministry of Agriculture and Rural Affairs, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China"},{"name":"School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, 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