{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T15:52:53Z","timestamp":1786117973981,"version":"build-2736575974"},"reference-count":78,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T00:00:00Z","timestamp":1785974400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005046","name":"Natural Science Foundation of Heilongjiang Province","doi-asserted-by":"publisher","award":["LH2023C029"],"award-info":[{"award-number":["LH2023C029"]}],"id":[{"id":"10.13039\/501100005046","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Rana dybowskii is an economically important amphibian in Northeast China, valued for its medicinal oviduct (Oviductus Ranae). However, the low proportion of females in artificial culture severely restricts industrial productivity. To elucidate the molecular mechanisms of hormone-induced sex reversal, based on our previous research results, we exposed tadpoles to 17\u03b2-estradiol (E2) or testosterone propionate (TP). We then monitored gonadal histology, conducted multi-stage transcriptome sequencing, and validated key genes via qRT-PCR. The results show that induction treatment with 40 \u03bcg\u00b7L\u22121 E2 and 80 \u03bcg\u00b7L\u22121 TP resulted in 93.33% female and 100% male populations, respectively. Both hormones promoted germ cell proliferation at the undifferentiated stage and directed stable ovarian or testicular development without intersex abnormalities. Ovarian differentiation involved progressive multi-wave transcriptional reprogramming, whereas testicular differentiation exhibited a concentrated gene expression burst during gonadal maturation. Steroid hormone biosynthesis, Wnt, and PPAR pathways formed the core regulatory network. Three female-biased genes (3\u03b1-hsd, Adcy3, Rspo1) and three male-biased genes (Ptgs2, Sox9, Shbg) were identified, with their expression dynamics aligned with histological progression. This study defines the optimal parameters and molecular signatures of bidirectional sex reversal in R. dybowskii. It provides a practical foundation for sex-control breeding and offers amphibian-based evidence for the conservation and plasticity of vertebrate sex determination.<\/jats:p>","DOI":"10.3390\/ani16152444","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T14:45:47Z","timestamp":1786027547000},"page":"2444","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular Mechanisms of Gonadal Differentiation Induced by 17\u03b2-Estradiol and Testosterone Propionate in Rana dybowskii"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-8614-2284","authenticated-orcid":false,"given":"Hualin","family":"Fu","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanqiu","family":"Sun","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiwen","family":"Sun","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangyu","family":"Meng","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-7584-2644","authenticated-orcid":false,"given":"Wei","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5498-3300","authenticated-orcid":false,"given":"Zhiheng","family":"Du","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,6]]},"reference":[{"key":"ref_1","first-page":"4739450","article-title":"Traditional Uses, Bioactive Constituents, Biological Functions, and Safety Properties of Oviductus ranae as Functional Foods in China","volume":"2019","author":"Zhang","year":"2019","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"103399","DOI":"10.1016\/j.aqrep.2026.103399","article-title":"Temperature influences sex determination in the giant spiny frog (Quasipaa spinosa) via alteration of DNA methylation and transcriptional profiles","volume":"46","author":"Zhu","year":"2026","journal-title":"Aquac. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1002\/jez.616","article-title":"The mechanism of sex determination in vertebrates\u2014Are sex steroids the key-factor?","volume":"313","author":"Nakamura","year":"2010","journal-title":"J. Exp. Zool. Part A Ecol. Genet. Physiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"5476","DOI":"10.1073\/pnas.082121499","article-title":"Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses","volume":"99","author":"Hayes","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_5","first-page":"187","article-title":"Endocrine Disruption in Aquatic Vertebrates","volume":"Volume 1163","author":"Vaudry","year":"2009","journal-title":"Trends in Comparative Endocrinology and Neurobiology"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ospina-Alvarez, N., and Piferrer, F. (2008). Temperature-Dependent Sex Determination in Fish Revisited: Prevalence, a Single Sex Ratio Response Pattern, and Possible Effects of Climate Change. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0002837"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1530\/REP-12-0048","article-title":"Differential effects of testosterone and 17\u03b2-estradiol on gonadal development in five anuran species","volume":"144","author":"Piprek","year":"2012","journal-title":"Reproduction"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/S0044-8486(01)00589-0","article-title":"Endocrine sex control strategies for the feminization of teleost fish","volume":"197","author":"Piferrer","year":"2001","journal-title":"Aquaculture"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.ygcen.2009.04.016","article-title":"Stress hormones mediate environment-genotype interactions during amphibian development","volume":"164","author":"Denver","year":"2009","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1093\/icb\/icp027","article-title":"Hormones and acoustic communication in anuran amphibians","volume":"49","author":"Leary","year":"2009","journal-title":"Integr. Comp. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.yhbeh.2005.05.011","article-title":"Historical perspective: Hormonal regulation of behaviors in amphibians","volume":"48","author":"Moore","year":"2005","journal-title":"Horm. Behav."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1051\/rnd:2004062","article-title":"Sex determination: The amphibian models","volume":"44","author":"Eggert","year":"2004","journal-title":"Reprod. Nutr. Dev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"739624","DOI":"10.1016\/j.aquaculture.2023.739624","article-title":"Sexual differentiation and sex reversal in tilapia (Oreochromis niloticus) by hormone 17 \u03b1 methyltestosterone similar to that used in cultivation systems","volume":"574","author":"Silva","year":"2023","journal-title":"Aquaculture"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"736450","DOI":"10.1016\/j.aquaculture.2021.736450","article-title":"Breeding of YY super-male of blotched snakehead (Channa maculata) and production of all-male hybrid (Channa argus \u2640 \u00d7 C. Maculata \u2642)","volume":"538","author":"Zhao","year":"2021","journal-title":"Aquaculture"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1006\/gcen.2001.7699","article-title":"Effect of Sex Steroids on Gonadal Differentiation and Sex Reversal in the Frog, Rana curtipes","volume":"124","author":"Saidapur","year":"2001","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1002\/jez.2037","article-title":"A Threshold Dosage of Testosterone for Female-to-Male Sex Reversal in Rana rugosa Frogs","volume":"325","author":"Oike","year":"2016","journal-title":"J. Exp. Zool. Part A Ecol. Genet. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Bosco, G., Kukoly, L.M., Porter, S.R., Jordan, D.C., Murphy, H.A., Knytl, M., Shaidani, N., Thomas, W.R., Anderson, C., and Dworkin, I. (2026). Sex-specific functional evolution of Dmrt1 in African clawed frogs (Xenopus), and the importance of genetic tipping points in developmental biology. PLoS Genet., 22.","DOI":"10.1371\/journal.pgen.1011992"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1186\/s12958-025-01377-0","article-title":"FOXL2 drives the differentiation of supporting gonadal cells in early ovarian development","volume":"23","author":"Danti","year":"2025","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1007\/s10126-024-10313-z","article-title":"A Novel Investigation for Early Sex Determination in Alive Adult European Seabass (Dicentrarchus labrax) Using a, dmrt1a, and dmrt1b Genes Expression in Tail Fin tissues","volume":"26","author":"Elmaghraby","year":"2024","journal-title":"Mar. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.ijbiomac.2022.06.098","article-title":"Function of Foxl2 and Dmrt1 proteins during gonadal differentiation in the olive flounder Paralichthys olivaceus","volume":"215","author":"Shu","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"102791","DOI":"10.1016\/j.aqrep.2025.102791","article-title":"Expression of sex-related genes and effects of dmrt1 siRNA-silencing on sex differentiation in juvenile yellow perch","volume":"42","author":"Xie","year":"2025","journal-title":"Aquac. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Mustapha, U.F., Peng, Y.-X., Huang, Y.-Q., Assan, D., Zhi, F., Shi, G., Huang, Y., Li, G.-L., and Jiang, D.-N. (2022). Comparative transcriptome analysis of the differentiating gonads in Scatophagus argus. Front. Mar. Sci., 9.","DOI":"10.3389\/fmars.2022.962534"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Tao, W., Chen, J., Tan, D., Yang, J., Sun, L., Wei, J., Conte, M.A., Kocher, T.D., and Wang, D. (2018). Transcriptome display during tilapia sex determination and differentiation as revealed by RNA-Seq analysis. BMC Genom., 19.","DOI":"10.1186\/s12864-018-4756-0"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Bachtrog, D., Mank, J.E., Peichel, C.L., Kirkpatrick, M., Otto, S.P., Ashman, T.L., Hahn, M.W., Kitano, J., Mayrose, I., and Ming, R. (2014). Sex Determination: Why So Many Ways of Doing It?. PLoS Biol., 12.","DOI":"10.1371\/journal.pbio.1001899"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1038\/nrg.2017.60","article-title":"Vertebrate sex determination: Evolutionary plasticity of a fundamental switch","volume":"18","author":"Capel","year":"2017","journal-title":"Nat. Rev. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"103161","DOI":"10.1016\/j.aqrep.2025.103161","article-title":"Histological observations and transcriptome analyses reveal the gonadal changes in blotched snakehead (Channa maculata) during sex reversal induced by 17\u03b2-estradiol","volume":"45","author":"Ou","year":"2025","journal-title":"Aquac. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.chemosphere.2018.11.092","article-title":"Testicular transcriptome alterations in zebrafish (Danio rerio) exposure to 17\u03b2-estradiol","volume":"218","author":"Zheng","year":"2019","journal-title":"Chemosphere"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"738984","DOI":"10.1016\/j.aquaculture.2022.738984","article-title":"Comparative transcriptome analysis of early sexual differentiation in the male and female gonads of common carp (Cyprinus carpio)","volume":"563","author":"Wang","year":"2023","journal-title":"Aquaculture"},{"key":"ref_29","first-page":"110467","article-title":"Effects of 17\u03b2-estradiol on early gonadal development and expression of genes implicated in sexual differentiation of a South American teleost, Astyanax altiparanae","volume":"248\u2013249","author":"Doretto","year":"2020","journal-title":"Comp. Biochem. Physiol. Part B Biochem. Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.ygeno.2018.11.008","article-title":"Dimorphism of sex and gonad-development-related genes in male and female lined seahorse, Hippocampus erectus, based on transcriptome analyses","volume":"111","author":"Qin","year":"2019","journal-title":"Genomics"},{"key":"ref_31","first-page":"100949","article-title":"Transcriptomic analysis of pituitary in female and male spotted scat (Scatophagus argus) after 17\u03b2-estradiol injection","volume":"41","author":"Shi","year":"2022","journal-title":"Comp. Biochem. Physiol. Part D Genom. Proteom."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"102373","DOI":"10.1016\/j.aqrep.2024.102373","article-title":"Characterization of early gonadal differentiation and estrogen-induced feminization in Chinese longsnout catfish (Leiocassis longirostris)","volume":"39","author":"Xiong","year":"2024","journal-title":"Aquac. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/S0044-8486(02)00205-3","article-title":"High density effects on the growth, development and survival of Litoria aurea tadpoles","volume":"215","author":"Browne","year":"2003","journal-title":"Aquaculture"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.aquatox.2012.10.003","article-title":"Effects of exposure to 17\u03b1-ethynylestradiol during larval development on growth, sexual differentiation, and abundances of transcripts in the liver of the wood frog (Lithobates sylvaticus)","volume":"126","author":"Tompsett","year":"2013","journal-title":"Aquat. Toxicol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.aquatox.2016.07.001","article-title":"Impaired gonadal and somatic development corroborate vulnerability differences to the synthetic estrogen ethinylestradiol among deeply diverged anuran lineages","volume":"177","author":"Tamschick","year":"2016","journal-title":"Aquat. Toxicol."},{"key":"ref_36","first-page":"183","article-title":"A Simplified Table for Staging Anuran Embryos and Larvae with Notes on Identification","volume":"16","author":"Gosner","year":"1960","journal-title":"Herpetologica"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Li, B., and Dewey, C.N. (2011). RSEM: Accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform., 12.","DOI":"10.1186\/1471-2105-12-323"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1080\/20014091111785","article-title":"Estrogen and Related Compounds: Biphasic Dose Responses","volume":"31","author":"Calabrese","year":"2001","journal-title":"Crit. Rev. Toxicol."},{"key":"ref_41","first-page":"00878","article-title":"Androgens and Notch signaling cooperate in seminiferous epithelium to regulate genes related to germ cell development and apoptosis","volume":"24","author":"Lustofin","year":"2024","journal-title":"Reprod. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1111\/j.1365-2109.2009.02286.x","article-title":"Effects of oestradiol-17\u03b2 or 17\u03b1-methyltestosterone administration on gonadal differentiation of largemouth bass Micropterus salmoides (Lacep\u00e8de)","volume":"40","author":"Arslan","year":"2009","journal-title":"Aquac. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.aquaculture.2017.05.037","article-title":"Establishment of monosex female production of sablefish (Anoplopoma fimbria) through direct and indirect sex control","volume":"479","author":"Luckenbach","year":"2017","journal-title":"Aquaculture"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1530\/JOE-19-0309","article-title":"PGE2 inhibits spermatogonia differentiation in zebrafish: Interaction with Fsh and an androgen","volume":"244","author":"Crespo","year":"2020","journal-title":"J. Endocrinol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"29467","DOI":"10.3390\/ijms161226186","article-title":"Protein Arginine Methyltransferase 6 Involved in Germ Cell Viability during Spermatogenesis and Down-Regulated by the Androgen Receptor","volume":"16","author":"Luo","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1002\/jat.1682","article-title":"A review of studies on androgen and estrogen exposure in fish early life stages: Effects on gene and hormonal control of sexual differentiation","volume":"31","author":"Leet","year":"2011","journal-title":"J. Appl. Toxicol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.ygcen.2014.06.009","article-title":"Global gene expression during early differentiation of Xenopus (Silurana) tropicalis gonad tissues","volume":"214","author":"Haselman","year":"2015","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Tang, Y., Chen, J.-Y., Ding, G.-H., and Lin, Z.-H. (2021). Analyzing the gonadal transcriptome of the frog Hoplobatrachus rugulosus to identify genes involved in sex development. BMC Genom., 22.","DOI":"10.1186\/s12864-021-07879-6"},{"key":"ref_49","first-page":"101586","article-title":"Gonadal sex differentiation in Eleutheronema tetradactylum: Histological features and transcriptomic insights from mature gonads","volume":"56","author":"Mkulo","year":"2025","journal-title":"Comp. Biochem. Physiol. Part D Genom. Proteom."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"149579","DOI":"10.1016\/j.gene.2025.149579","article-title":"Transcriptomic analysis of the differences in gene expression between testis and ovary of Dybowski\u2019s frog (Rana dybowskii) in reproduction period","volume":"962","author":"Zhang","year":"2025","journal-title":"Gene"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1387\/ijdb.130192rp","article-title":"Sexual dimorphism of AMH, DMRT1 and RSPO1 localization in the developing gonads of six anuran species","volume":"57","author":"Piprek","year":"2013","journal-title":"Int. J. Dev. Biol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"5445","DOI":"10.1038\/s41598-020-62408-x","article-title":"The transcriptome of the newt Cynops orientalis provides new insights into evolution and function of sexual gene networks in sarcopterygians","volume":"10","author":"Biscotti","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1007\/s11427-021-2075-x","article-title":"Sex determination mechanisms and sex control approaches in aquaculture animals","volume":"65","author":"Li","year":"2022","journal-title":"Sci. China Life Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1152\/physrev.00044.2019","article-title":"Sex determination, gonadal sex differentiation, and plasticity in vertebrate species","volume":"101","author":"Nagahama","year":"2020","journal-title":"Physiol. Rev."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1159\/000328823","article-title":"Dimorphic Expression of Various Transcription Factor and Steroidogenic Enzyme Genes during Gonadal Ontogeny in the Air-Breathing Catfish, Clarias gariepinus","volume":"5","author":"Raghuveer","year":"2011","journal-title":"Sex. Dev."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"113893","DOI":"10.1016\/j.ygcen.2021.113893","article-title":"Role of sex steroids in fish sex determination and differentiation as revealed by gene editing","volume":"313","author":"Zhou","year":"2021","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0044-8486(95)01075-0","article-title":"Hormonal induction of sex reversal in fish","volume":"138","author":"Pandian","year":"1995","journal-title":"Aquaculture"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1071\/RD09027","article-title":"Involvement of peroxisome proliferator-activated receptor? In gonadal steroidogenesis and steroidogenic acute regulatory protein expression","volume":"21","author":"Kowalewski","year":"2009","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"4641","DOI":"10.2174\/1381612811319250010","article-title":"The Biology of the Peroxisome Proliferator-activated Receptor System in the Female Reproductive Tract","volume":"19","author":"Velez","year":"2013","journal-title":"Curr. Pharm. Des."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1278","DOI":"10.1093\/hmg\/ddn036","article-title":"R-spondin1 plays an essential role in ovarian development through positively regulating Wnt-4 signaling","volume":"17","author":"Tomizuka","year":"2008","journal-title":"Hum. Mol. Genet."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1007\/s11248-009-9247-2","article-title":"Goat RSPO1 over-expression rescues sex-reversal in Rspo1-knockout XX mice but does not perturb testis differentiation in XY or sex-reversed XX mice","volume":"18","author":"Buscara","year":"2009","journal-title":"Transgenic Res."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"113788","DOI":"10.1016\/j.ygcen.2021.113788","article-title":"Knockdown of R-spondin1 leads to partial sex reversal in genetic female Chinese soft-shelled turtle Pelodiscus sinensis","volume":"309","author":"Zhang","year":"2021","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"2190","DOI":"10.1007\/s10528-024-10815-4","article-title":"Gonadal Transcriptome Analysis Reveals that SOX17 and CYP26A1 are Involved in Sex Differentiation in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis)","volume":"63","author":"Wan","year":"2025","journal-title":"Biochem. Genet."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1042\/bj3510067","article-title":"Human 3\u03b1-hydroxysteroid dehydrogenase isoforms (AKR1C1\u2013AKR1C4) of the aldo-keto reductase superfamily: Functional plasticity and tissue distribution reveals roles in the inactivation and formation of male and female sex hormones","volume":"351","author":"Penning","year":"2000","journal-title":"Biochem. J."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/S0306-4522(03)00287-2","article-title":"Gender differences in the regulation of 3\u03b1-hydroxysteroid dehydrogenase in rat brain and sensitivity to neurosteroid-mediated stress protection","volume":"120","author":"Mitev","year":"2003","journal-title":"Neuroscience"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"pgad055","DOI":"10.1093\/pnasnexus\/pgad055","article-title":"cAMP controls the balance between dormancy and activation of primordial follicles in mouse ovaries","volume":"2","author":"Zheng","year":"2023","journal-title":"PNAS Nexus"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.repbre.2021.02.004","article-title":"Role of sox family genes in teleostean reproduction-an overview","volume":"1","author":"Anitha","year":"2021","journal-title":"Reprod. Breed."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1530\/REP-10-0200","article-title":"Isolation of sox9 duplicates in catfish: Localization, differential expression pattern during gonadal development and recrudescence, and hCG-induced up-regulation of sox9 in testicular slices","volume":"140","author":"Raghuveer","year":"2010","journal-title":"Reproduction"},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Li, X., Yu, H., Wang, Y., Liu, X., Liu, Y., Qu, J., and Wang, X. (2018). Roles of Two Sox9 Genes during Gonadal Development in Japanese Flounder: Sex Differentiation, Spermatogenesis and Gonadal Function Maintenance. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020512"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1095\/biolreprod.106.053124","article-title":"Constitutive Expression of Prostaglandin-Endoperoxide Synthase 2 by Somatic and Spermatogenic Cells Is Responsible for Prostaglandin E2 Production in the Adult Rat Testis1","volume":"76","author":"Winnall","year":"2007","journal-title":"Biol. Reprod."},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Szybiak-Skora, W., Cyna, W., and Lacka, K. (2025). New Insights in the Diagnostic Potential of Sex Hormone-Binding Globulin (SHBG)\u2014Clinical Approach. Biomedicines, 13.","DOI":"10.3390\/biomedicines13051207"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"4781","DOI":"10.1038\/s41467-024-49025-2","article-title":"A candidate sex determination locus in amphibians which evolved by structural variation between X- and Y-chromosomes","volume":"15","author":"Kuhl","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1242\/dev.048751","article-title":"Opposite roles of DMRT1 and its W-linked paralogue, DM-W, in sexual dimorphism of Xenopus laevis: Implications of a ZZ\/ZW-type sex-determining system","volume":"137","author":"Yoshimoto","year":"2010","journal-title":"Development"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"740479","DOI":"10.1016\/j.aquaculture.2023.740479","article-title":"Production of neofemale by 17\u03b2-estradiol and YY super-male breeding in mandarin fish (Siniperca chuatsi)","volume":"581","author":"Liu","year":"2024","journal-title":"Aquaculture"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1007\/s11427-014-4797-9","article-title":"Genetic basis and biotechnological manipulation of sexual dimorphism and sex determination in fish","volume":"58","author":"Mei","year":"2015","journal-title":"Sci. China Life Sci."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1002\/dvg.22720","article-title":"Simple and efficient CRISPR\/Cas9-mediated targeted mutagenesis in Xenopus tropicalis","volume":"51","author":"Nakayama","year":"2013","journal-title":"Genesis"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"112636","DOI":"10.1016\/j.mce.2025.112636","article-title":"Epigenetic regulation of sex: The role of DNA methylation and zbtb38 in zebrafish sex differentiation and heat-induced masculinization","volume":"609","author":"Pierron","year":"2025","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.ygcen.2013.11.013","article-title":"Environmental estrogens inhibit growth of rainbow trout (Oncorhynchus mykiss) by modulating the growth hormone-insulin-like growth factor system","volume":"196","author":"Hanson","year":"2014","journal-title":"Gen. Comp. Endocrinol."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/15\/2444\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T14:51:50Z","timestamp":1786027910000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/15\/2444"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,6]]},"references-count":78,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ani16152444"],"URL":"https:\/\/doi.org\/10.3390\/ani16152444","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,6]]}}}