Figure 3. Regulation of prostaglandin synthesis by Lh and Tnfα in brown trout preovulatory follicles.

(a) In vitro effects of sLh (25 ng/mL) and rTnfα (50 ng/mL) on the expression of enzymes involved in prostaglandin synthesis. The relative expression of ptgs1 and ptgs2 was determined by qPCR and normalized to the abundance of 18s. (b) Determination of PGF_2α production in vitro by intact ovarian follicles treated with sLh. Trout follicles were incubated with sLh (25 ng/mL) in the absence or presence of TAPI-1 (50 μM) and INDO (10 μg/mL) for 16 h at 15 °C and PGF_2α was measured in the incubation media. (c) PGF_2α production in response to rTnfα treatment in vitro. Follicles were incubated with rTnfα (1, 10 and 50 ng/mL) or rTnfα (50 ng/mL) with INDO (10 μg/ml) for 16 h at 15 °C. (d) PGE₂ production by brown trout preovulatory ovarian follicles. Follicles were incubated in the absence or presence of sLh (25 ng/mL) for 16 h at 15 °C and PGE₂ levels were measured in the incubation media. In all graphs, data are the mean ± SEM (n = 3–7). *P < 0.05, with respect to control; ^#P < 0.05, with respect to sLh (b) and with respect to rTnfα (50 ng/mL) (c).