NR2F2 regulation of interstitial cell fate in the embryonic mouse testis and its impact on differences of sex development [ChIP-Seq]

Testicular fetal Leydig cells are specialized for androgen production during embryogenesis. Testosterone is essential for regulating sex differentiation, spermatogenesis, and fertility. Deficiencies in Leydig cell differentiation can lead to various disorders of sex development and male reproductive conditions, such as ambiguous genitalia, hypospadias, cryptorchidism, and infertility. Understanding the differentiation of fetal Leydig cells is essential for comprehending male sexual differentiation, reproductive health, and fertility. Fetal Leydig cells originate from proliferating progenitor cells in the gonadal interstitium, marked by genes like Arx, Pdgfra, Tcf21, Wnt5a, and Nr2f2 (COUP-TFII). However, the precise mechanisms governing the transition from interstitial cells to Leydig cells remain elusive. Through integrated approaches involving animal models and multiomics, we have demonstrated that fetal Leydig cells originate from a COUP-TFII positive non-steroidogenic interstitial cell population. COUP-TFII promotes progenitor cell fate while suppressing Leydig cell differentiation. Moreover, embryonic deletion of COUP-TFII in mouse testes resulted in disorders of sex development, including reduced testicular size, Leydig cell hypoplasia, cryptorchidism, and hypospadias. Collectively, our findings highlight the critical role of COUP-TFII in orchestrating the transition from interstitial cells to Leydig cells during testicular development. Testes from E14.5 CD-1 (Charles River stock number 022) were separated from the mesonephros, snap-frozen, and stored at −80 °C. Two independent ChIP-seq experiments were performed by Active Motif, using 18 μg of sheared chromatin from 30 pooled embryonic testes and 5 μl of COUP-TFII antibody (Active motif, 61213). Libraries were sequenced by ActiveMotif as single-end 75-mers on an Illumina NextSeq 500 instrument.