Supplementary Materials Supplemental Data supp_292_43_17577__index. the testicular cord and aberrant testis development. However, the underlying molecular mechanism was unclear. In this study, we found that constitutive activation of in Sertoli cells led to ectopic expression of the granulosa cell-specific marker FOXL2 in testes. Co-staining experiments revealed that FOXL2-positive cells were derived from Sertoli cells, and Sertoli cells were transformed into granulosa-like cells after overactivation. Further studies exhibited that CTNNB1 induced expression by directly binding to transcription factor Tcf/Lef-binding sites in the promoter region. We also found that direct overexpression of decreased the appearance of Sertoli cell-specific genes in principal Sertoli cells. Used together, these outcomes show that repression of -catenin (CTNNB1) signaling is necessary for lineage maintenance of Sertoli cells. Our research provides a brand-new system for Sertoli cell lineage maintenance during gonad advancement. is vital for directing Sertoli cell differentiation in XY gonads (3,C5). In XX gonads, which absence appearance, the somatic cell differentiates into granulosa cell, which is certainly regulated with the RSPO1/WNT4–catenin (CTNNB1) signaling pathway (6,C8). Inactivation of and before sex perseverance results in incomplete female-to-male sex reversal in mice (8,C11). In comparison, overactivation of before sex perseverance using triggered male-to-female sex reversal with an elevated expression of FOXL2 and reduced expression of SOX9 in the male gonad (12). Recent studies found that the differentiated Sertoli cells and granulosa cells have the potential to mutually transform after sex commitment. FOXL2 is usually a forkhead transcription factor specifically expressed in ovarian granulosa cells (13, 14), and deletion of results in aberrant ovarian follicle development and the dysgenesis of ovaries (13). Interestingly, it has been exhibited that FOXL2 is also required for granulosa cell lineage maintenance. Inactivation of in the granulosa cells of adult ovaries results in an up-regulation of the testis-specific gene and the transformation of granulosa cells into Sertoli-like cells along with the formation of a testicular cord-like structure (15). The gene encodes a nuclear transcription factor, which is usually abundantly expressed in Sertoli cells. Deletion Prostaglandin E1 kinase activity assay of causes the reprogramming of Sertoli cells to granulosa-like cells postnatally, which in turn prospects to dysgenesis of the testes (16). Our previous study (17) found that constitutive activation of by deletion of exon 3 in Sertoli cells after sex determination using transgenic mice caused testicular cord disruption and the loss of appearance of Sertoli cell-specific genes. Nevertheless, the root molecular mechanism continues to be unclear. Oddly enough, in today’s study, we discovered that the granulosa cell-specific marker Prostaglandin E1 kinase activity assay FOXL2 was portrayed in the remnant testicular cords of overactivated mice ectopically. Lineage tracing tests uncovered that Sertoli cells had been changed into granulosa-like cells after overactivation. Further research confirmed that CTNNB1 induced appearance in the Sertoli cell series by directly getting together with T cell aspect/lymphoid enhancer aspect (Tcf/Lef)4-binding sites in the promoter area. These total outcomes indicate that repression of WNT/-catenin signaling is vital for Sertoli cell lineage maintenance, and activation of causes an up-regulation of FOXL2, which leads towards the change of Sertoli Rabbit Polyclonal to ALS2CR11 cells into granulosa-like cells. Outcomes Ectopic appearance of FOXL2 proteins in the testes of Ctnnb1+/flox(ex girlfriend or boyfriend3) AMH-Cre mice Our prior studies discovered that overactivation of by deletion of exon 3 in Sertoli cells using transgenic mice triggered testicular cable disruption and lack of Sertoli cell-specific genes’ appearance (17). To explore the explanation for unusual testis advancement in mice further, the expression of Sertoli cell-specific and granulosa cell-specific genes was Prostaglandin E1 kinase activity assay examined by real-time and immunostaining PCR assays. In charge testes, CTNNB1 proteins was localized on the plasma membrane of Sertoli cells and germ cells from E13.5 to P1 (supplemental Fig. S1, testes, the deposition of CTNNB1 protein in the nucleus of Sertoli cells was first observed at E14.5 (supplemental Fig. S1was overactivated in Sertoli cells, which was consistent with the findings obtained in our earlier study (17). As demonstrated in supplemental Fig. S2, testes from E13.5, the WT1-positive cells were still observed in the periphery region of testicular cords at E15.5 and E17.5 (supplemental Fig. S2, testes (supplemental Fig. 2and testes at E15.5 (supplemental Fig. S2testes (supplemental Fig. S2testes at E15.5 and E17.5 (supplemental Fig. S2, and testes at E14.5 (Fig. 1and testes at E13.5 were SOX9-positive (supplemental Fig. S3testes (supplemental Fig. S3mice. IHC was carried out using anti-FOXL2 antibody. FOXL2 protein (mice at E13.5 (probably caused Sertoli to granulosa-like cells transformation. To test this hypothesis, a double staining experiment was performed. WT1 was specifically indicated in the Sertoli cells of control testes as demonstrated in Fig. 2, and testes (Fig. 2and testes were most likely derived from WT1-positive Sertoli cells. To further confirm this effect, the GFP-expressed (mice to generate mice (18). In control mice (and is specifically indicated in Sertoli cells. In the mice, a FOXL2 transmission.