Sp1 regulates steroidogenic genes and LHCGR expression in primary human luteinized granulosa cells

Abstract

Luteinizing hormone and human chorionic gonadotropin (hCG) bind to the luteinizing hormone/ chorionic gonadotropin receptor (LHCGR). LHCGR is required to maintain corpus luteum function but the mechanisms involved in the regulation of LHCGR in human luteal cells remain incompletely understood. This study aimed to characterize the expression of LHCGR mRNA in primary human luteinized granulosa cells (hLGCs) obtained from patients undergoing in vitro fertilization and to correlate LHCGR expression with the response of hLGCs to hCG by assessing the expression of genes known to be markers of hCG actions. The results show that LHCGR expression is low in freshly isolated cells but recovers rapidly in culture and that hCG maintains LHCGR expression, suggesting a positive feedback loop. The activity of a LHCGR-LUC reporter increased in cells treated with hCG but not with follicle-stimulating hormone. Treatment with hCG also stimulated the expression of genes involved in steroidogenesis in a time-dependent manner. LHCGR promoter expression was found to be regulated by SP1, which we show is highly expressed in hLGCs. Moreover, SP1 inhibition prevented the stimulation of steroidogenic genes and the increase in LHCGR-LUC reporter activity by hCG. Finally, we provide evidence that a complex formed by SP1 and GATA4 may play a role in the maintenance of LHCGR expression. This report reveals the mechanisms involved in the regulation of the LHCGR and provides experimental data demonstrating that the proximal region of the LHCGR promoter is sufficient to drive the expression of this gene in primary hLGCs.

Spatiotemporal expression pattern of miR-205, miR-26a-5p, miR-17-5p, let-7b-5p, and their target genes during different stages of corpus luteum in Egyptian buffaloes

“…Meanwhile, the P4 concentration reached its peak during CLM, whereas the concentration was decreased at CLH and CLA. Supporting this notion, the freshly isolated luteinized granulosa cells showed a reduction in LHCGR and then recovered by hCG [ 52 ], which resulted in lower P4 production. Moreover, the high levels of LHCGR in the current study were offset by a reduction in miR-205-5p and vice versa, which leads us to lay the cornerstone for understanding the regulatory mechanism of LHCGR and P4 secretion.…”

Comprehensive analysis of mRNAs and miRNAs in the ovarian follicles of uniparous and multiple goats at estrus phase

“…2e, f). Approximately 37% of the DEgenes in the Uni-S vs Mul-S group and 41% in the Uni-L vs Mul-L group were reported to be associated with mammalian reproduction, such as TNFAIP6, MMP9, INSL3, LEPR, 3BHSD, LHCGR, ARL4C, CD36, CYP11A1, AMDHD1, SPOCK2, AMDHD1, MFAP5, CCL21, PTGFR, and SERPINA5 [23,[47][48][49][50][51][52][53][54][55][56][57][58][59][60]. Of these genes, TNFAIP6, CYP11A1, CD36, PTGFR, and SER-PINA5 were found to be associated with the ovulation rate.…”

Correlation Networks Provide New Insights into the Architecture of Testicular Steroid Pathways in Pigs

“…It was already showed in other species that SP1 and NR5A1 are involved in the regulation of transcriptional expression of STAR [ 60 , 61 ] and CYP11A1 [ 60 ]. SP1 has been described as being able to regulate the LHCGR expression [ 62 ] and NR5A1 to participate to the regulation of the expression of CYP19A1 [ 63 ]. The involvement of CREB1 in the control of steroidogenesis was already described in human ovaries [ 64 ].…”