Single-cell sequencing reveals transcriptional dynamics regulated by ERα in mouse ovaries

Abstract

Abstract Estrogen receptor α (ERα) is a crucial regulator of reproductive function, mediating the diverse effects of estrogen on the development and function of the ovary. However, the molecular mechanisms underlying ERα-regulated transcriptional dynamics in ovarian cells remain poorly understood. In the present study, we generated ERα knockout (αERKO) mice through CRISPR/Cas9 strategy. Then we employed single-cell RNA sequencing technology (scRNA-seq) to profile the transcriptomes of individual cells in wild type (WT) and αERKO mouse ovaries to uncover novel insights into ERα-regulated transcriptional change and ovarian cellular heterogeneity. Using several bioinformatics analyses, our study identified distinct cell populations within the ovarian tissue, including 5 distinct cell clusters associated with oocytes and 7 with granulosa cells. We further conducted differential gene expression analysis to explore the transcriptional profiles of these cell populations in response to ERα deletion. Our findings revealed that ERα deletion led to the dysregulation of genes involved in ovarian infertility, ovulation cycle, and steroidogenesis in oocytes and granulosa cells, indicating that ERα plays a central role in modulating the transcriptional landscape of ovarian cells. Consequently, we confirmed that Greb1, a prototypical ERα target gene, was induced by ERα binding to ERE upstream of the Greb1 promoter in granulosa cells. Additionally, we uncovered a complex network of signaling pathways that were significantly modulated by ERα, implicating their involvement in the regulation of ovarian function and ovulation process. In conclusion, our study offers an in-depth, single-cell resolution of ERα-regulated transcriptional dynamics in mouse ovaries, enhancing our understanding of ERα's role in female reproductive biology and providing potential targets for novel therapeutic interventions for ovarian disorders.