BEEF SPECIES-RUMINANT NUTRITION CACTUS BEEF SYMPOSIUM: Maternal immune modulation prior to embryo arrival in the uterus is important for establishment of pregnancy in cattle1.

In 1953, Sir Peter Medawar first recognized the allogeneic properties of a developing conceptus and rationalized that an "immune-tolerant" physiological state must exist during pregnancy. Early theories speculated that the conceptus evaded the maternal immune system completely, but 40 yr after Medawar's observations, Wegmann proposed that the maternal immune system shifts the cytokine profile away from inflammatory cytokine production when an embryo is present. The economic consequences and production losses of subfertile animals have been well documented in studies evaluating calving distribution. Despite advances in understanding infertility or subfertility, few technologies exist to identify subfertile animals or improve fertility beyond hormonal intervention associated with synchronization protocols. Work in rodents and some livestock species indicates that the uterine immune cell population shifts dramatically after copulation and these early immune-modulated events establish a receptive uterine environment. Clearly, as evident in embryo transfer, the presence of a conceptus is sufficient to establish communication for pregnancy establishment but does not rule out the importance of other physiological events to prime the maternal immune system prior to blastocyst arrival in the uterus. In support of this concept, work in our laboratory and by others has demonstrated that autologous intrauterine transfer of peripheral immune cells prior to embryo transfer can increase pregnancy rates and accelerate conceptus development in women and cattle. Understanding aberrant immune regulation in subfertile animals may provide markers for subfertility or targets for clinical intervention to enhance fertility, particularly when using reproductive technologies.

ASAS-SSR Triennial Reproduction Symposium: Looking Back and Moving Forward-How Reproductive Physiology has Evolved: WNTs role in bovine folliculogenesis and estrogen production.

Appreciation of mechanisms that affect steroidogenesis is critical to identifying compromising signals that may decrease reproductive efficiency. Follicle maturation and steroidogenesis requires coordinated actions from the pituitary gonadotropins and local ovarian signaling molecules. ß-Catenin (CTNNB1), the lynchpin molecule of canonical wingless-type mouse mammary tumor virus integration site (WNT) signaling, is required for maximal gonadotropin stimulation of steroid production from granulosa (GC) and luteal cells. WNTs are locally secreted glycoproteins involved in ovarian development and folliculogenesis. In cultured bovine GC, WNT2 and AKT mRNAs and CTNNB1 protein increase after FSH stimulation. Likewise, CTNNB1 protein is greater in large antral follicles with high intrafollicular estradiol concentrations, suggesting the hormonal milieu responsible for increased estradiol content modulates CTNNB1 accumulation. In addition, concurrent treatment of FSH and WNT3A in GC results in reduced steroidogenic enzymes and ovarian differentiation factors. It is likely that FSH regulation of WNT signaling establishes a negative feedback loop to ensure CTNNB1 remains controlled. To explore the mechanism resulting in this inhibitory effect, AKT pathway modulators were utilized and unveiled a requirement for AKT activity in FSH-mediated CTNNB1 accumulation. Cells treated with FSH, IGF-1, and IGF-1 + FSH had increased CTNNB1 protein accumulation compared with controls. Similarly, estradiol medium concentrations increased in treated cells compared with non-treated controls, while co-treatment of FSH and IGF-1 with the AKT inhibitor LY294002 reduced CTNNB1 and estradiol production. Subsequent studies evaluated whether FSH regulation of CTNNB1 occurs through a specific phosphorylation event. In bovine GC, phosphorylation of CTNNB1 at Ser-552 was demonstrated in FSH-treated cells, whereas IGF-1 treatment did not phosphorylate CTNNB1 Ser-552. Data indicate that in cattle phosphorylation on CTNNB1 Ser-552 is a protein kinase A (PKA) dependent, protein kinase B (AKT) independent event. Data suggest that CTNNB1 regulated by AKT is a fundamental component of FSH-induced estrogen production. However, AKT's role in estradiol synthesis does not appear to be through phosphorylation of CTNNB1 Ser-552. The complex interplay between FSH and ovarian WNT/CTNNB1 signaling is key to regulation of follicle maturation and steroidogenesis.