Single-Nucleus Transcriptomic Atlas of Goat (Capra hircus) Ovarian Prolificacy

Molecular mechanisms of follicular atresia and prolificacy of mammal remain unclear. We surveyed the single-cell transcriptomic landscape of ovaries from single and prolificacy goat and identified seven ovarian cell types with distinct gene-expression, transcriptional factor networks and reciprocal interactions signatures. In-depth dissection of gene-expression dynamics of granulosa cells (GCs) that displayed development stage-specific expression patterns and specific gene signatures were identified that may reflect developmental competency and ovarian reserve. what's more, we revealed the origin of theca cells. Further analysis of cell-type-specific prolificacy-associated transcriptional changes uncovered apoptosis, anabolism and response to hormone stimulation as are crucial factor in dominant follicle development and ovulation. Additionally, differentially expressed genes (DEGs) of SERPINE2 can interact with CYP19A1 to promote cell proliferation, inhibit apoptosis and promoting the anabolism were observed in mouse granulosa cells. Thus, our work provides a comprehensive understanding of the cell-type-specific mechanisms underlying goat ovarian prolificacy at single-cell resolution, provides key insights into offers important clues for improving follicle recruitment in vivo and revealing new diagnostic biomarkers and potential therapeutic targets for ovulation disorder. Overall design: Granulosa cells were isolated from C57BL/6J mouse ovarian tissue 3 days after PMSG induction. SV40 gene was transfected into cells by lentivirus transfection to obtain immortalized function. Briefly, the GCs were isolated from the follicular fluid using density gradient centrifugation at 2000 rpm for 30 min. Following centrifugation, the sediments layer was collected and washed with Dulbecco phosphate-buffered saline (GIBCO). After a brief exposure to 0.2% hyaluronidase (Sigma) at 37?, the cell suspensions were centrifuged at 3000 rpm for 5 min. Then, the purity of isolated GCs was assessed by immunofluorescence for GC markers. All cells were cultured in DMEM/F12 medium (GIBCO) supplemented with 10% FBS (GIBCO), 100 U/ml penicillin, and 100 mg/ml streptomycin (GIBCO) at 37? in 5% CO2. All the cell cultures were tested negative for myco- plasma contamination.