FIGURE 3.

Gonadotropin secretion and gene expression in primary cultures of anterior pituitary cells treated with CB NPs. Rat primary anterior pituitary cells were cultured for 24 h with increasing concentrations of CB NPs (25–100 μg/mL). At the end of the incubation, FSH and LH concentrations in the culture media of primary pituitary cells were determined by ELISA. FSH/LH ratio is also shown in panel (A). Total RNAs were extracted and Fshb, Lhb, Cga (B) and Follistatin (C) mRNA levels were determined by real-time quantitative PCR. Data are expressed as percentage over untreated cells and expressed as means ± SEM from 4 independent experiments. Statistical differences were determined by non-parametric Kruskal-Wallis test followed by Dunnet’s multiple comparison test. *p < 0.05 and **p < 0.01 compared to no CB NPs.