{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T19:50:21Z","timestamp":1793044221524,"version":"4.5.0"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,16]],"date-time":"2022-09-16T00:00:00Z","timestamp":1663286400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["25292118 16H04970 L19549"],"award-info":[{"award-number":["25292118 16H04970 L19549"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009037","name":"Science and Technology Research Partnership for Sustainable Development","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009037","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:p>Japanese flounder (<jats:italic>Paralichthys olivaceus<\/jats:italic>) is an important marine fish species of both fisheries and aquaculture in Northeast Asia. The commercial interest for all-female progenies due to several sex-related traits has prompted basic research on the mechanisms of sex determination in this species. By conducting a linkage analysis of the sex-determining locus, we initially identified 12 microsatellite markers linked to sex in 11 scaffolds, whose localization was restricted to a specific region of linkage group 9. Sequence analysis of this region identified 181 genes based on the UniProt database annotations. Among them, the <jats:italic>amh<\/jats:italic> gene was considered a potential candidate for sex determination because this gene is known to have taken over the role of sex determination in many teleosts. An in-depth sequence analysis of both the coding and non-coding regions of <jats:italic>amh<\/jats:italic> in XX and XY individuals detected nine SNPs linked with maleness. However, because these substitutions were synonymous, the upstream and downstream regions of <jats:italic>amh<\/jats:italic> were also investigated and a male-specific variant with deletions in the promoter region was detected. This truncated Y-specific <jats:italic>amh<\/jats:italic> variant was named <jats:italic>amhy<\/jats:italic>, and the <jats:italic>amh<\/jats:italic> shared by both sexes was named <jats:italic>amhx<\/jats:italic>. The association analysis using both females and males of the genotypic sex inferred by the presence\/absence of <jats:italic>amhy<\/jats:italic> found complete association with phenotypic sex and genotype. Gene expression analysis in larvae derived from a single-pair progeny by quantitative real-time PCR detected <jats:italic>amhy<\/jats:italic> transcripts in the larval trunks between 20 and 100 days after hatching only in XY larvae. Localization of <jats:italic>amhy<\/jats:italic> by <jats:italic>in situ<\/jats:italic> hybridization was detected in presumptive Sertoli cells of XY gonads. Expression of <jats:italic>amhx<\/jats:italic> was almost undetectable in both XX and XY genotypes. Loss of Amh function by CRISPR-Cas9 induced male-to-female sex reversal, indicating that this gene was necessary for the masculinization of XY individuals. In conclusion, the complete linkage of <jats:italic>amhy<\/jats:italic> with males, its early expression in XY gonads before testicular differentiation, and the induction of sex reversal by loss-of-function mutation support the view that <jats:italic>amhy<\/jats:italic> is the sex-determining gene in this species.<\/jats:p>","DOI":"10.3389\/fgene.2022.1007548","type":"journal-article","created":{"date-parts":[[2022,9,16]],"date-time":"2022-09-16T08:25:18Z","timestamp":1663316718000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":44,"title":["Y-specific amh allele, amhy, is the master sex-determining gene in Japanese flounder Paralichthys olivaceus"],"prefix":"10.3389","volume":"13","author":[{"given":"Ricardo Shohei","family":"Hattori","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keiichiro","family":"Kumazawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masatoshi","family":"Nakamoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Nakano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshiya","family":"Yamaguchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takeshi","family":"Kitano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eiichi","family":"Yamamoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanako","family":"Fuji","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Sakamoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,16]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. 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