Transcriptomic alterations in the ovine caruncular endometrium due to imbalanced nutrition and FSH-induced ovarian hyperstimulation

Background: Imbalanced diet and exogenous gonadotrophins affect uterine function and morphology. In sheep, FSH-induced superovulation alters implantation-related gene expression, influenced by both treatment and diet. In this study, we used deep RNA sequencing (NGS, RNA-Seq) to expand our understanding of these effects on the caruncular endometrium. Methods: Ewes (n = 3-5/group) were separated into control fed (CF), overfed (OF), and underfed (UF) groups, with each group subdivided between FSH (superovulated; SOV) or saline (negative controls; CONT) treatment. Caruncular samples were collected on day 10 of diestrus of the subsequent estrous cycle, with samples from CF_CONT also collected on day 5 to assess time-dependent changes. Results: The 1484 differentially expressed genes (DEGs, P < 0.01, FDR < 0.05) identified between CF_CONT animals at days 5 and 10 were predominantly associated with increased immune activity and cellular metabolic processes and cellular proliferation. In CONT animals, imbalanced nutrition (i.e., both OF and UF) was associated with enrichment of terms associated with cell adhesion and differentiation, immune response and angiogenesis. The FSH carry-over effects resulted in a higher number of DEGs in CF animals (1374), than in OF (168) or UF (18), mostly associated with dysregulation of cell cycle and hormonal sensitivity. Conclusion: The absence of genes concurrently affected by superovulation (SOV) in all feeding regimes indicates that the effects of FSH on the caruncular transcriptome are multidirectional and dependent upon body condition. Therefore, the homeostasis of ovine caruncles is influenced by both body condition and superovulation (SOV), potentially affecting uterine receptivity.