Figure 5 A-C. Effect of metformin on FSHR mRNA abundance in granulosa cells.
Granulosa cells from prehierarchical follicles (3–5 and 6–8 mm) and preovulatory follicles (9–12 mm) were dispersed into to single-cell suspension and treated with 0, 1, 10, or 20 mM metformin for 180 min in the presence or absence of rhFSH (10 ng/ml) at 41°C. Control received no rhFSH and no metformin treatments. Total RNA extracted from granulosa cell and reverse transcribed. Approximately 100 ng of cDNA was used in quantitative PCR to quantify FSHR mRNA or RPL19 mRNA in separate reactions. Each reaction was run in duplicates, and the critical threshold (CT) values were averaged, subtracted from that of RPL19 mRNA, and converted from log-linear to linear terms. Data were standardized to RPL19 mRNA and expressed as fold-difference versus control. A, B, C P < 0.001; n=4 replicates of 4 broiler breeder hens each. Inset: Photograph of prehierarchical follicles and the most-recently recruited preovulatory follicle to denote the source of granulosa cell (open box).