{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:12:16Z","timestamp":1789085536424,"version":"build-2803163510"},"reference-count":46,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,11]],"date-time":"2022-12-11T00:00:00Z","timestamp":1670716800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Makowski Family Endowment","award":["63470698"],"award-info":[{"award-number":["63470698"]}]},{"name":"Makowski Family Endowment","award":["0222933"],"award-info":[{"award-number":["0222933"]}]},{"name":"Gonadotropin Research Fund","award":["63470698"],"award-info":[{"award-number":["63470698"]}]},{"name":"Gonadotropin Research Fund","award":["0222933"],"award-info":[{"award-number":["0222933"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In the mouse, two distinct populations of Leydig cells arise during testis development. Fetal Leydig cells arise from a stem cell population and produce T required for masculinization. It is debated whether they persist in the adult testis. A second adult Leydig stem cell population gives rise to progenitor-immature-mature adult type Leydig cells that produce T in response to LH to maintain spermatogenesis. In testis of adult null male mice lacking either only LH (Lhb\u2212\/\u2212) or LHR (Lhr\u2212\/\u2212), mature Leydig cells are absent but fetal Leydig cells persist. Thus, it is not clear whether other ligands signal via LHRs in Lhb null mice or LH signals via other receptors in the absence of LHR in Lhr null mice. Moreover, it is not clear whether truncated LHR isoforms generated from the same Lhr gene promoter encode functionally relevant LH receptors. To determine the in vivo roles of LH-LHR signaling pathway in the Leydig cell lineage, we generated double null mutant mice lacking both LH Ligand and all forms of LHR. Phenotypic analysis indicated testis morpho-histological characteristics are identical among double null and single mutants which all showed poorly developed interstitium with a reduction in Leydig cell number and absence of late stage spermatids. Gene expression analyses confirmed that the majority of the T biosynthesis pathway enzyme-encoding mRNAs expressed in Leydig cells were all suppressed. Expression of thrombospondin-2, a fetal Leydig cell marker gene was upregulated in single and double null mutants indicating that fetal Leydig cells originate and develop independent of LH-LHR signaling pathway in vivo. Serum and intratesticular T levels were similarly suppressed in single and double mutants. Consequently, expression of AR-regulated genes in Sertoli and germ cells were similarly affected in single and double mutants without any evidence of any additive effect in the combined absence of both LH and LHR. Our studies unequivocally provide genetic evidence that in the mouse testis, fetal Leydig cells do not require LH-LHR signaling pathway and a one-to-one LH ligand-LHR signaling pathway exists in vivo to regulate adult Leydig cell lineage and spermatogenesis.<\/jats:p>","DOI":"10.3390\/ijms232415725","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T01:42:27Z","timestamp":1670809347000},"page":"15725","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Lhb\u2212\/\u2212Lhr\u2212\/\u2212 Double Mutant Mice Phenocopy Lhb\u2212\/\u2212 or Lhr\u2212\/\u2212 Single Mutants and Display Defects in Leydig Cells and Steroidogenesis"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4894-9744","authenticated-orcid":false,"given":"Zhenghui","family":"Liu","sequence":"first","affiliation":[{"name":"Division of Reproductive Sciences, Department of Obstetrics and Gynecology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2800-3487","authenticated-orcid":false,"given":"Mark","family":"Larsen","sequence":"additional","affiliation":[{"name":"Division of Reproductive Sciences, Department of Obstetrics and Gynecology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenmin","family":"Lei","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Louisville School of Medicine, Louisville, KY 40202, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. V.","family":"Rao","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Molecular and Human Genetics, Obstetrics and Gynecology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL 33199, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T. Rajendra","family":"Kumar","sequence":"additional","affiliation":[{"name":"Division of Reproductive Sciences, Department of Obstetrics and Gynecology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA"},{"name":"Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,11]]},"reference":[{"key":"ref_1","first-page":"1581","article-title":"Gonadotropins: Chemistry and biosynthesis","volume":"Volume I","author":"Neill","year":"2006","journal-title":"Knobil\u2019s Physiology of Reproduction"},{"key":"ref_2","unstructured":"Strauss, J.F., and Barbieri, R.L. (2019). Yen & Jaffe\u2019s Reproductive Endocrinology, Elsevier. [8th ed.]."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Simoni, M., and Huhtaniemi, I. (2017). Gonadotropins. 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