Single-Cell Sequencing Reveals an Intrinsic Heterogeneity of the Preovulatory Follicular Microenvironment

The follicular microenvironment, including intra-follicular granulosa cells (GCs), is responsible for oocyte maturation and subsequent ovulation. However, the functions of GCs and cellular components of the follicular microenvironment in preovulatory follicles have not been extensively explored. Here, we surveyed the single-cell transcriptome of the follicular microenvironment around MII oocytes in six human preovulatory follicles in in vitro fertilization. There were six different cell types in the preovulatory follicles, including GCs and various immune cells. In GCs, we identified nine different functional clusters with different functional transcriptomic profiles, including specific clusters involved in inflammatory responses and adhesive function. Follicular macrophages are involved in immune responses, extracellular matrix remoulding and assist GCs in promoting the oocyte meiotic resumption. Interestingly, we observed that the specific terminal state subcluster of GCs with high levels of adhesive-related molecules should result in macrophage recruitment and residence, further contributing to an obvious heterogeneity of the immune cell proportion in preovulatory follicles from different patients. Our results provide a comprehensive understanding of the transcriptomic landscape of the preovulatory follicular microenvironment at the single-cell level. It provides valuable insights into understanding the regulation of the oocyte maturation and ovulation process, offering potential clues for the diagnosis and treatment of oocyte-maturation-related and ovulation-related diseases.

Automated summary

This study provides a comprehensive understanding of the follicular microenvironment at the single-cell level. The functions and cellular components of the follicular microenvironment, including granulosa cells (GCs) and immune cells, are explored. GCs are found to have different functional clusters involved in inflammatory responses and adhesive function, while follicular macrophages play roles in immune responses and extracellular matrix remodeling, assisting GCs in promoting oocyte maturation. The specific subclusters of GCs with high levels of adhesive-related molecules contribute to the recruitment and residence of macrophages, leading to heterogeneity in the proportion of immune cells in preovulatory follicles from different patients.