{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T03:56:38Z","timestamp":1788494198737,"version":"build-2803163510"},"posted":{"date-parts":[[2026,1,30]]},"group-title":"Evolutionary Biology","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T00:00:00Z","timestamp":1769731200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"European Research Council","award":["101045439, 3D-REVOLUTION"],"award-info":[{"award-number":["101045439, 3D-REVOLUTION"]}]},{"award":["PID2022-143253NB-I00\/AEI\/10.13039\/501100011033\/"],"award-info":[{"award-number":["PID2022-143253NB-I00\/AEI\/10.13039\/501100011033\/"]}],"id":[{"id":"https:\/\/ror.org\/003x0zc53","id-type":"ROR","asserted-by":"publisher"}]},{"award":["RYC2023-045620-I"],"award-info":[{"award-number":["RYC2023-045620-I"]}],"id":[{"id":"https:\/\/ror.org\/003x0zc53","id-type":"ROR","asserted-by":"publisher"}]},{"name":"EMBO","award":["EMBO ALTF 537-2020"],"award-info":[{"award-number":["EMBO ALTF 537-2020"]}]},{"name":"NIH","award":["1S10RR027867"],"award-info":[{"award-number":["1S10RR027867"]}]},{"name":"NIH","award":["R01HD103064"],"award-info":[{"award-number":["R01HD103064"]}]},{"name":"NSF","award":["1854642"],"award-info":[{"award-number":["1854642"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,30]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Sex determination in vertebrates can be initiated by a wide range of genetic or environmental triggers. Yet, the degree to which gonadal cell types and genetic programs are conserved remains unresolved. Here we employed single-cell transcriptomics to characterize the temperature-dependent sex determination (TSD) program in gonads from the turtle species\n                  <jats:italic>Trachemys scripta.<\/jats:italic>\n                  Comparative analyses against species with genetic sex determination, like mouse (XY) and chicken (ZW), revealed a marked divergence in cell type repertoires and functions during vertebrate evolution. Unlike mammals, fetal Leydig cells are absent from the early gonads of\n                  <jats:italic>T. scripta<\/jats:italic>\n                  , where the supporting lineage expresses genes required for androgen synthesis. Evolutionary reconstructions show that this lineage derives from a\n                  <jats:italic>Pax2<\/jats:italic>\n                  -positive mesenchymal population, suggesting an ancestral condition in Archelosauria that differs from the primarily coelomic epithelium origin in the mammalian clade. Transcriptional dynamics and co-expression analyses revealed the recruitment of lineage-specific transcription factors, including\n                  <jats:italic>Twist1 or Runx1<\/jats:italic>\n                  , into the genetic programs of vertebrate clades. Our findings reveal extensive plasticity of the cellular and genetic mechanisms of vertebrate sex determination and suggest that this flexibility is a key feature of gonadal evolution.\n                <\/jats:p>","DOI":"10.64898\/2026.01.29.701242","type":"posted-content","created":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T15:00:16Z","timestamp":1769785216000},"source":"Crossref","is-referenced-by-count":2,"title":["Flexibility of cell fates and functions across sex determination systems revealed by comparative single-cell analyses"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4120-0140","authenticated-orcid":false,"given":"Rafael D.","family":"Acemel","sequence":"first","affiliation":[{"name":"Centro Andaluz de Biolog\u00eda del Desarrollo (CABD), Consejo Superior de Investigaciones Cient\u00edficas\/Universidad Pablo de Olavide\/Junta de Andaluc\u00eda, Sevilla, Spain"},{"name":"Max-Delbr\u00fcck Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetics and Sex Development Group, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2621-4272","authenticated-orcid":false,"given":"Boris","family":"Tezak","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Duke University Medical Center, Durham, NC 27710"},{"name":"Biology Department, Wesleyan University, Middletown, CT 06459"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vicky Wai Yee","family":"Chung","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetics and Sex Development Group, Berlin, Germany"},{"name":"Current address: Institute of Molecular and Clinical Ophthalmology, Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6267-704X","authenticated-orcid":false,"given":"Alicia","family":"Hurtado","sequence":"additional","affiliation":[{"name":"Centro Andaluz de Biolog\u00eda del Desarrollo (CABD), Consejo Superior de Investigaciones Cient\u00edficas\/Universidad Pablo de Olavide\/Junta de Andaluc\u00eda, Sevilla, Spain"},{"name":"Max-Delbr\u00fcck Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetics and Sex Development Group, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scarlett","family":"Albertson","sequence":"additional","affiliation":[{"name":"Biology Department, Wesleyan University, Middletown, CT 06459"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8541-0110","authenticated-orcid":false,"given":"Ceri J.","family":"Weber","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Duke University Medical Center, Durham, NC 27710"},{"name":"Current address: Department of Cell and Developmental Biology, University of California San Diego, La Jolla, CA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6587-0969","authenticated-orcid":false,"given":"Blanche","family":"Capel","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Duke University Medical Center, Durham, NC 27710"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3165-036X","authenticated-orcid":false,"given":"Dar\u00edo G.","family":"Lupi\u00e1\u00f1ez","sequence":"additional","affiliation":[{"name":"Centro Andaluz de Biolog\u00eda del Desarrollo (CABD), Consejo Superior de Investigaciones Cient\u00edficas\/Universidad Pablo de Olavide\/Junta de Andaluc\u00eda, Sevilla, Spain"},{"name":"Max-Delbr\u00fcck Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetics and Sex Development Group, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026020513043306000_2026.01.29.701242v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg.2017.60"},{"key":"2026020513043306000_2026.01.29.701242v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/348448A0"},{"key":"2026020513043306000_2026.01.29.701242v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/348450a0"},{"key":"2026020513043306000_2026.01.29.701242v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/nature08298"},{"key":"2026020513043306000_2026.01.29.701242v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.ygcen.2006.11.001"},{"key":"2026020513043306000_2026.01.29.701242v1.6","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.cellbio.042308.13350"},{"key":"2026020513043306000_2026.01.29.701242v1.7","doi-asserted-by":"publisher","DOI":"10.15252\/embr.201540667"},{"key":"2026020513043306000_2026.01.29.701242v1.8","doi-asserted-by":"crossref","first-page":"1589","DOI":"10.1016\/j.celrep.2018.01.043","article-title":"Deciphering 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