Global analysis of FSH‐regulated gene expression and histone modification in mouse granulosa cells

Abstract

Follicle-stimulating hormone (FSH) regulates ovarian follicular development through a specific gene expression program. We analyzed FSH-regulated transcriptome and histone modification in granulosa cells during follicular development. We used super-stimulated immature mice and collected granulosa cells before and 48 h after stimulation with equine chorionic gonadotropin (eCG). We profiled the transcriptome using RNA-sequencing (N = 3/time-point) and genome-wide trimethylation of lysine 4 of histone H3 (H3K4me3; an active transcription marker) using chromatin immunoprecipitation and sequencing (ChIP-Seq; N = 2/time-point). Across the mouse genome, 14,583 genes had an associated H3K4me3 peak and 63-66% of these peaks were observed within ≤1 kb promoter region. There were 72 genes with differential H3K4me3 modification at 48 h eCG (absolute log fold change > 1; false discovery rate [FDR] < 0.05) relative to 0 h eCG. Transcriptome data analysis showed 1463 differentially expressed genes at 48 h eCG (absolute log fold change > 1; FDR < 0.05). Among the 20 genes with differential expression and altered H3K4me3 modification, Lhcgr had higher H3K4me3 abundance and expression, while Nrip2 had lower H3K4me3 abundance and expression. Using ChIP-qPCR, we showed that FSH-regulated expression of Lhcgr, Cyp19a1, Nppc, and Nrip2 through regulation of H3K4me3 at their respective promoters. Transcript isoform analysis using Kallisto-Sleuth tool revealed 875 differentially expressed transcripts at 48 h eCG (b > 1; FDR < 0.05). Pathway analysis of RNA-seq data demonstrated that TGF-β signaling and steroidogenic pathways were regulated at 48 h eCG. Thus, FSH regulates gene expression in granulosa cells through multiple mechanisms namely altered H3K4me3 modification and inducing specific transcripts. These data form the basis for further studies investigating how these specific mechanisms regulate granulosa cell functions.

Follicle-stimulating hormone mediates the consumption of serum-derived glycogen by bovine cumulus-oocyte complexes during in vitro maturation

“…In this work, the effect of FSH on the expansion of cumulus cells, estradiol production, and oocyte nuclear maturation was observed ex vivo, thus reinforcing the seminal effects of this hormone on cellular hallmarks of oocyte maturation [32][33][34]. Cumulus cells mediate FSH signaling pathways that contribute to the acquisition of developmental competence [14,[35][36][37].…”

Janus Kinase 3 phosphorylation and the JAK/STAT pathway are positively modulated by follicle-stimulating hormone (FSH) in bovine granulosa cells

“…A more accurate determination of the number and function of FSH-regulated genes as well as LH-regulated genes have been reported over the years 15,16,61-63 and demonstrate the importance of functional studies during the later stages of follicular development to better coordinate the ovarian activity. In this regard, we have shown previously the suppression of JAK3 in vivo by endogenous LH or hCG injection in bovine species and identified JAK3 binding partners in GC using the yeast two-hybrid method 15,17 .…”

Cellular atlas of the human ovary using morphologically guided spatial transcriptomics and single-cell sequencing

“…Likewise, the list of granulosa-specific genes included canonical marker genes HSD17B1, INHBB, FOXL2, and AMH and the recently reported TNNI3, MAGED2, CD99, SERPINE2, CDH2, and BEX1 (9, 13), along with many histone protein-coding genes (data S5), which have not been reported for granulosa cells. However, numerous studies have suggested that proliferating granulosa cells undergo large-scale epigenetic change during late folliculogenesis, partly driven by gonadotrophins such FSH (40)(41)(42)(43). The list also included genes involved in granulosa cell-oocyte signaling, such as the transzonal projection-related genes FSCN1 and MYO10, and the tetraspanin protein gene TSPAN7, which is involved in extracellular vesiclemediated signaling between granulosa cells and the oocyte (44,45).…”