Figure 1. Mediatory effects of Tnfα and PGF_2α on sLh-induced contraction of brown trout preovulatory follicles.

(a) Effects of sLh on follicle contraction. Punctured follicles were incubated for 16 h at 15 °C with epinephrine (EPI; 10 μM), sLh (25 ng/mL), sLh plus TAPI-1 (50 μM) and sLh plus INDO (10 μg/mL). The results are expressed as percent change with respect to the unpunctured control group that was set at 100%. (b) Effects of rTnfα and PGF_2α on follicle contraction. Punctured preovulatory follicles were incubated for 16 h at 15 °C with rTnfα (1, 10 and 50 ng/mL), rTnfα (50 ng/mL) in the presence of INDO (10 μg/mL) and PGF_2α (0.2, 2 and 20 μg/mL). In (a) and (b), data are expressed as percent change with respect to the unpunctured control group that was set at 100%. (c) Relative expression of tnfα and adam17 analyzed by qPCR in ovarian follicles incubated in the absence or presence of sLh (25 ng/mL). (d) Ovarian Tnfα secretion in brown trout follicle incubates by Western blot. Preovulatory follicles were incubated with sLh (25 ng/mL) in the absence or presence of TAPI-1 (50 μM) for 16 h at 15 °C. A representative Western blot is shown in the inset and the full-length blot is presented in Supplementary Figure S1. In all graphs, data are the mean ± standard error (SEM) (n = 3–5). *P < 0.05, with respect to control; ^#P < 0.05, with respect to sLh (a,d) and with respect to rTnfα (50 ng/mL) (b).