Stromal-Derived Factor-1 (SDF-1) directly promotes genes expressed within the ovulatory cascade.

LINARI, M; PELUFFO, MC; ROJO, JL; YOUNG, KA

CABA

Congreso; Reunión Conjunta de Biociencias 2017; 2017

SAIC y 9 sociedades mas

SDF1, a chemokine that is expressed in multiple tissue types including the ovary, serves as the ligand to chemokine CXC-motif receptor-4 (CXCR4). While LH upregulates follicular CXCR4, the role that SDF1/CXCR4 signaling might play in the COC (cumulus-oocyte complex) was unknown. We hypothesized that the SDF1/CXCR4 system was present in feline COCs and that COCs culture with SDF1 would directly upregulate genes expressed in the ovulatory cascade (HAS2, TSG6, AREG, and GDF9). To address this hypothesis, a COC culture was established. Ovaries (n= 50) were obtained from adult domestic cats during breeding season and COCs were mechanically recovered from antral follicles. The culture environment was optimized by testing different conditions (media, serums, serum concentrations and pretreatments). During culture optimization, COCs (n=24) were cultured at different time-points (3, 12 and 24h) with and without gonadotropins (GNT). Then, the effects of 25ng/ml SDF1 and CXCR-4 inhibitor were examined in the COC at all time-points (n=48) and gene expression was assessed by real-time PCR. CXCR4 and SDF1 immunofluorescence was also performed. MEM-hepes with 1% of charcoal stripped-FBS was the optimum medium for this culture, as seen by the expansion of the COC at 24h. The mRNA expression of HAS2, TSG6, and AREG peaked at 3h in the GNT group as compared to GNT treatment in the 12 and 24h groups, and as compared to negative controls (p