{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,19]],"date-time":"2026-10-19T15:56:37Z","timestamp":1792425397252,"version":"4.5.0"},"reference-count":39,"publisher":"The Endocrine Society","issue":"2","license":[{"start":{"date-parts":[[2025,1,6]],"date-time":"2025-01-06T00:00:00Z","timestamp":1736121600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["MOP-81387"],"award-info":[{"award-number":["MOP-81387"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fondation du CHU de Qu\u00e9bec\u2013Universit\u00e9 Laval"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Leydig cells produce hormones that are required for male development, fertility, and health. Two Leydig cell populations produce these hormones but at different times during development: fetal Leydig cells, which are active during fetal life, and adult Leydig cells, which are functional postnatally. Historically, our ability to understand the origin and function of Leydig cells has been made difficult by the lack of genetic models to exclusively target these cells. Taking advantage of the Leydig cell\u2013exclusive expression pattern of the Insl3 gene, we used a CRISPR\/Cas9 gene-editing strategy to knock-in iCre recombinase into the mouse Insl3 locus. To demonstrate the Leydig cell\u2013exclusive nature of our iCre line, lineage-tracing experiments were performed by crossing Insl3iCre mice with a Rosa26LoxSTOPLox-TdTomato reporter. iCre activity was restricted to male offspring. TdTomato fluorescence was detected both in fetal and adult Leydig cells and colocalized with CYP17A1, a classic Leydig cell marker. Prior to birth, fluorescence was observed in fetal Leydig cells beginning at embryonic day 13.0. Fluorescence was also detected in adult Leydig cells starting at postnatal day 5 and continuing to the mature testis. Fluorescence was not detected in any other fetal or adult tissue examined, except for the unexpected finding that the adrenal cortex contains some Insl3-expressing Leydig-like cells. Our Leydig cell\u2013exclusive iCre line therefore constitutes an invaluable new tool to study not only the origin of Leydig cells but also to target genes that have been long-proposed to be important for the development and functioning of these critical endocrine cells.<\/jats:p>","DOI":"10.1210\/endocr\/bqaf012","type":"journal-article","created":{"date-parts":[[2025,1,17]],"date-time":"2025-01-17T20:02:10Z","timestamp":1737144130000},"source":"Crossref","is-referenced-by-count":5,"title":["A New Leydig Cell\u2013Exclusive Cre Line Allows Lineage Tracing of Fetal and Adult Leydig Cell Populations in the Mouse"],"prefix":"10.1210","volume":"166","author":[{"given":"Nicholas M","family":"Robert","sequence":"first","affiliation":[{"name":"Reproduction, Mother and Child Health, Centre de recherche du centre hospitalier universitaire de Qu\u00e9bec\u2013Universit\u00e9 Laval , Qu\u00e9bec City, QC G1V 4G2 ,","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shirley","family":"Ferrier-Tarin","sequence":"additional","affiliation":[{"name":"Reproduction, Mother and Child Health, Centre de recherche du centre hospitalier universitaire de Qu\u00e9bec\u2013Universit\u00e9 Laval , Qu\u00e9bec City, QC G1V 4G2 ,","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4738-6284","authenticated-orcid":false,"given":"Jacques J","family":"Tremblay","sequence":"additional","affiliation":[{"name":"Reproduction, Mother and Child Health, Centre de recherche du centre hospitalier universitaire de Qu\u00e9bec\u2013Universit\u00e9 Laval , Qu\u00e9bec City, QC G1V 4G2 ,","place":["Canada"]},{"name":"Centre de recherche en Reproduction, D\u00e9veloppement et Sant\u00e9 Interg\u00e9n\u00e9rationnelle, Department of Obstetrics, Gynecology, and Reproduction, Faculty of Medicine, Universit\u00e9 Laval , Qu\u00e9bec City, QC G1V 0A6 ,","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"80","published-online":{"date-parts":[[2025,1,17]]},"reference":[{"issue":"3","key":"2025020410362258700_bqaf012-B1","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1038\/10364","article-title":"Cryptorchidism in mice mutant for Insl3","volume":"22","author":"Nef","year":"1999","journal-title":"Nat Genet"},{"issue":"5","key":"2025020410362258700_bqaf012-B2","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1210\/mend.13.5.0272","article-title":"Targeted disruption of the Insl3 gene causes bilateral cryptorchidism","volume":"13","author":"Zimmermann","year":"1999","journal-title":"Mol Endocrinol"},{"issue":"5","key":"2025020410362258700_bqaf012-B3","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1359\/jbmr.080204","article-title":"Mutations in the insulin-like factor 3 receptor are associated with osteoporosis","volume":"23","author":"Ferlin","year":"2008","journal-title":"J Bone Miner Res"},{"issue":"11","key":"2025020410362258700_bqaf012-B4","doi-asserted-by":"crossref","first-page":"110935","DOI":"10.1016\/j.celrep.2022.110935","article-title":"Deciphering the origins and fates of steroidogenic lineages in the mouse testis","volume":"39","author":"Ademi","year":"2022","journal-title":"Cell Rep"},{"issue":"1","key":"2025020410362258700_bqaf012-B5","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1152\/physrev.1986.66.1.71","article-title":"Differentiation of mammalian embryonic gonad","volume":"66","author":"Byskov","year":"1986","journal-title":"Physiol Rev"},{"issue":"7","key":"2025020410362258700_bqaf012-B6","doi-asserted-by":"crossref","first-page":"2657","DOI":"10.1096\/fj.12-225060","article-title":"Dynamic changes in fetal Leydig cell populations influence adult Leydig cell populations in mice","volume":"27","author":"Barsoum","year":"2013","journal-title":"FASEB J"},{"issue":"1","key":"2025020410362258700_bqaf012-B7","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1095\/biolreprod.113.115790","article-title":"The nuclear receptor NR2F2 activates Star expression and steroidogenesis in mouse MA-10 and MLTC-1 Leydig cells","volume":"91","author":"Mendoza-Villarroel","year":"2014","journal-title":"Biol Reprod"},{"key":"2025020410362258700_bqaf012-B8","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/978-1-59745-453-7_4","volume-title":"The Leydig Cell in Health and Disease","author":"Ge","year":"2007","edition":"2nd ed"},{"issue":"7","key":"2025020410362258700_bqaf012-B9","doi-asserted-by":"crossref","first-page":"E1227","DOI":"10.1210\/jc.2013-4199","article-title":"Human testicular peritubular cells host putative stem Leydig cells with steroidogenic capacity","volume":"99","author":"Landreh","year":"2014","journal-title":"J Clin Endocrinol Metab"},{"issue":"18","key":"2025020410362258700_bqaf012-B10","doi-asserted-by":"crossref","first-page":"E1924","DOI":"10.1073\/pnas.1320735111","article-title":"Fetal programming of adult leydig cell function by androgenic effects on stem\/progenitor cells","volume":"111","author":"Kilcoyne","year":"2014","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2025020410362258700_bqaf012-B11","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.semcdb.2021.05.007","article-title":"Stem Leydig cells: current research and future prospects of regenerative medicine of male reproductive health","volume":"121","author":"Li","year":"2022","journal-title":"Semin Cell Dev Biol"},{"issue":"2","key":"2025020410362258700_bqaf012-B12","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1530\/REP-10-0404","article-title":"Good planning and serendipity: exploiting the Cre\/Lox system in the testis","volume":"141","author":"Smith","year":"2011","journal-title":"Reproduction"},{"issue":"1","key":"2025020410362258700_bqaf012-B13","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1095\/biolreprod.109.078766","article-title":"Fetal testis dysgenesis and compromised Leydig cell function in Tgfbr3 (beta glycan) knockout mice","volume":"82","author":"Sarraj","year":"2010","journal-title":"Biol Reprod"},{"issue":"6","key":"2025020410362258700_bqaf012-B14","doi-asserted-by":"crossref","first-page":"2960","DOI":"10.1210\/endo.139.6.6046","article-title":"Relaxin-like factor expression as a marker of differentiation in the mouse testis and ovary","volume":"139","author":"Balvers","year":"1998","journal-title":"Endocrinology"},{"issue":"1","key":"2025020410362258700_bqaf012-B15","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1038\/nn.2467","article-title":"A robust and high-throughput Cre reporting and characterization system for the whole mouse brain","volume":"13","author":"Madisen","year":"2010","journal-title":"Nat Neurosci"},{"issue":"3","key":"2025020410362258700_bqaf012-B16","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1111\/andr.12601","article-title":"Phosphorylation of GATA4 serine 105 but not serine 261 is required for testosterone production in the male mouse","volume":"7","author":"Bergeron","year":"2019","journal-title":"Andrology"},{"issue":"19","key":"2025020410362258700_bqaf012-B17","doi-asserted-by":"crossref","first-page":"12009","DOI":"10.3390\/ijms231912009","article-title":"A short promoter region containing conserved regulatory motifs is required for steroidogenic acute regulatory protein (star) gene expression in the mouse testis","volume":"23","author":"Bouchard","year":"2022","journal-title":"Int J Mol Sci"},{"issue":"4","key":"2025020410362258700_bqaf012-B18","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1534\/genetics.119.301984","article-title":"Twenty-seven tamoxifen-inducible iCre-driver mouse strains for eye and brain, including seventeen carrying a new inducible-first constitutive-ready allele","volume":"211","author":"Korecki","year":"2019","journal-title":"Genetics"},{"issue":"10","key":"2025020410362258700_bqaf012-B19","doi-asserted-by":"crossref","first-page":"5002","DOI":"10.1210\/en.2012-1417","article-title":"Identification, proliferation, and differentiation of adult Leydig stem cells","volume":"153","author":"Stanley","year":"2012","journal-title":"Endocrinology"},{"issue":"5","key":"2025020410362258700_bqaf012-B20","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1083\/jcb.200409107","article-title":"Progenitor cells of the testosterone-producing Leydig cells revealed","volume":"167","author":"Davidoff","year":"2004","journal-title":"J Cell Biol"},{"issue":"3","key":"2025020410362258700_bqaf012-B21","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1038\/ng1003","article-title":"Requirement of Bmpr1a for Mullerian duct regression during male sexual development","volume":"32","author":"Jamin","year":"2002","journal-title":"Nat Genet"},{"issue":"9","key":"2025020410362258700_bqaf012-B22","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1002\/dvg.20231","article-title":"Development of a steroidogenic factor 1\/Cre transgenic mouse line","volume":"44","author":"Bingham","year":"2006","journal-title":"Genesis"},{"issue":"11","key":"2025020410362258700_bqaf012-B23","doi-asserted-by":"crossref","first-page":"1581","DOI":"10.1210\/me.2015-1200","article-title":"Fetal Leydig cells persist as an androgen-independent subpopulation in the postnatal testis","volume":"29","author":"Shima","year":"2015","journal-title":"Mol Endocrinol"},{"issue":"9","key":"2025020410362258700_bqaf012-B24","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1002\/dvg.20428","article-title":"Generation of Cyp17iCre transgenic mice and their application to conditionally delete estrogen receptor alpha (Esr1) from the ovary and testis","volume":"46","author":"Bridges","year":"2008","journal-title":"Genesis"},{"issue":"1","key":"2025020410362258700_bqaf012-B25","doi-asserted-by":"crossref","first-page":"e84541","DOI":"10.1371\/journal.pone.0084541","article-title":"Targeting of GFP-Cre to the mouse Cyp11a1 locus both drives cre recombinase expression in steroidogenic cells and permits generation of Cyp11a1 knock out mice","volume":"9","author":"O\u2019Hara","year":"2014","journal-title":"PLoS One"},{"issue":"2","key":"2025020410362258700_bqaf012-B26","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1002\/dvg.20266","article-title":"Human CYP11A1 promoter drives Cre recombinase expression in the brain in addition to adrenals and gonads","volume":"45","author":"Wu","year":"2007","journal-title":"Genesis"},{"issue":"4","key":"2025020410362258700_bqaf012-B27","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1002\/dvg.20198","article-title":"Inhibin alpha-iCre mice: Cre deleter lines for the gonads, pituitary, and adrenal","volume":"44","author":"Jorgez","year":"2006","journal-title":"Genesis"},{"issue":"6","key":"2025020410362258700_bqaf012-B28","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1093\/molehr\/3.6.459","article-title":"Relaxin-like factor: a highly specific and constitutive new marker for Leydig cells in the human testis","volume":"3","author":"Ivell","year":"1997","journal-title":"Mol Hum Reprod"},{"issue":"1\u20132","key":"2025020410362258700_bqaf012-B29","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.mce.2005.03.017","article-title":"Constitutive regulation of the Insl3 gene in rat Leydig cells","volume":"241","author":"Sadeghian","year":"2005","journal-title":"Mol Cell Endocrinol"},{"issue":"1","key":"2025020410362258700_bqaf012-B30","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/BF00225571","article-title":"Selective destruction and regeneration of rat Leydig cells in vivo. A new method for the study of seminiferous tubular-interstitial tissue interaction","volume":"242","author":"Kerr","year":"1985","journal-title":"Cell Tissue Res"},{"issue":"2","key":"2025020410362258700_bqaf012-B31","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1210\/endo-118-2-709","article-title":"Ethylene dimethanesulfonate destroys Leydig cells in the rat testis","volume":"118","author":"Morris","year":"1986","journal-title":"Endocrinology"},{"issue":"5","key":"2025020410362258700_bqaf012-B32","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1095\/biolreprod33.5.1213","article-title":"Specific destruction of Leydig cells in mature rats after in vivo administration of ethane dimethyl sulfonate","volume":"33","author":"Molenaar","year":"1985","journal-title":"Biol Reprod"},{"issue":"7","key":"2025020410362258700_bqaf012-B33","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1046\/j.1365-2443.1996.00254.x","article-title":"Identical origin of adrenal cortex and gonad revealed by expression profiles of Ad4BP\/SF-1","volume":"1","author":"Hatano","year":"1996","journal-title":"Genes Cells"},{"issue":"1","key":"2025020410362258700_bqaf012-B34","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1016\/j.ydbio.2006.07.030","article-title":"Identification of a novel population of adrenal-like cells in the mammalian testis","volume":"299","author":"Val","year":"2006","journal-title":"Dev Biol"},{"issue":"4","key":"2025020410362258700_bqaf012-B35","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1530\/REP-07-0133","article-title":"Expression of Cyp21a1 and Cyp11b1 in the fetal mouse testis","volume":"134","author":"Hu","year":"2007","journal-title":"Reproduction"},{"issue":"5","key":"2025020410362258700_bqaf012-B36","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/0022-3468(92)90466-K","article-title":"Benign testicular tumors in children with congenital adrenal hyperplasia","volume":"27","author":"Srikanth","year":"1992","journal-title":"J Pediatr Surg"},{"issue":"5","key":"2025020410362258700_bqaf012-B37","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1210\/jcem-70-5-1408","article-title":"Steroidogenic enzyme activities, morphology, and receptor studies of a testicular adrenal rest in a patient with congenital adrenal hyperplasia","volume":"70","author":"Clark","year":"1990","journal-title":"J Clin Endocrinol Metab"},{"issue":"1","key":"2025020410362258700_bqaf012-B38","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1083\/jcb.55.1.126","article-title":"A new crystal\u2013containing cell in human adrenal cortex","volume":"55","author":"Magalhaes","year":"1972","journal-title":"J Cell Biol"},{"issue":"6","key":"2025020410362258700_bqaf012-B39","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1111\/andr.13201","article-title":"Reinke crystals: hallmarks of adult Leydig cells in humans","volume":"10","author":"Planinic","year":"2022","journal-title":"Andrology"}],"container-title":["Endocrinology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/endo\/advance-article-pdf\/doi\/10.1210\/endocr\/bqaf012\/61489467\/bqaf012.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/endo\/article-pdf\/166\/2\/bqaf012\/61489467\/bqaf012.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/endo\/article-pdf\/166\/2\/bqaf012\/61489467\/bqaf012.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T10:37:04Z","timestamp":1738665424000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/endo\/article\/doi\/10.1210\/endocr\/bqaf012\/7959789"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,6]]},"references-count":39,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2025,1,6]]}},"URL":"https:\/\/doi.org\/10.1210\/endocr\/bqaf012","relation":{},"ISSN":["1945-7170"],"issn-type":[{"value":"1945-7170","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,2]]},"published":{"date-parts":[[2025,1,6]]},"article-number":"bqaf012"}}