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Developmental Expression of Bone Morphogenetic Protein 2 (BMP2)-Induced Genes in Fetal Mouse Ovaries [A335]

INTRODUCTION: Granulosa cells (GCs) and oocytes assemble to form the primordial follicle (PF) reserve. Reduced PF reserve results in consequential infertility and menopause-related disorders. Ovarian transcriptome analysis was done to understand the mechanism of bone morphogenetic proteins (BMPs) or estradiol-17β (E2)induced PF formation. METHODS: RNA from fetal (E16-E19) and postnatal (P1 and P2) mouse ovaries and E16 ovaries cultured with or without BMP2 or E2 for 24 hours were analyzed by RNA sequencing and bioinformatics. In vivo and in vitro expression of selected candidate genes and spatio-temporal expression of corresponding proteins were examined using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence localization, respectively. RESULTS: Remarkable increases in Gata2, Gata4, Egr2 and Hepsin mRNA were evident on E18 (one day prior to PF formation). Protein immunosignal was present in somatic cells (SC) but the intensity was higher in GCs on E18 through P1. BMP2 or E2 upregulated gene expression in vivo and in vitro; however, the effect of E2 was less for Gata4 and Egr2. BMP2 significantly upregulated all proteins in GCs, whereas E2 upregulated GATA2 and HEPSIN proteins in SCs as well. CONCLUSION: In summary, Gata2, Gata4, Egr2 and Hepsin mRNA levels increase around PF formation. Correspondingly, intensities of immunosignal increase in differentiating GCs. BMP2 or E2 may induce differentiation of ovarian SCs into GCs by cell-specific activation of Gata2, Gata4 and/or Egr2. Hepsin, a serine proteinase, may participate in ovary remodeling. We conclude that BMP2-induced GATA2, GATA4, EGR2 and Hepsin may regulate SC to GC transition, leading to the formation of PFs.