Effect of nanomolar concentration of Met-AEA and heparin on bull sperm capacitation.

<p>Spermatozoa were incubated for 45 min at 38.5°C with different treatments. Bars indicate the percentage of capacitated spermatozoa (pattern B) determined by CTC. (t = 0; t = 45: controls). Data are expressed as mean ± SEM. <b>A</b>: Met-AEA, Heparin or Met-AEA plus Heparin. ***p<0.001 vs. control (n = 6). <b>B</b>: Heparin, Heparin plus Ant-CB1 or Heparin plus Ant-TRPV1. *p<0.05 Heparin vs. control, Ant-CB1 and Ant-TRPV1 (n = 4).</p

Evaluation of CB1, CB2 and TRPV1 participation in bull sperm capacitation.

<p>Spermatozoa were incubated for 45 min at 38.5°C with different treatments. Bars indicate the percentage of capacitated spermatozoa (pattern B) determined by CTC. Data are expressed as mean ± SEM. <b>A</b>: sp-TALP (control), Met-AEA (1.4 nM), Met-AEA plus Ant-CB1 (CB1 antagonist SR141716A; 0.1 nM), Met-AEA plus Ant-CB2 (CB2 antagonist SR144528); **p<0.01 Met-AEA vs. control and Met-AEA plus Ant-CB1 (n = 6). **p<0.01 Met-AEA plus Ant-CB2 vs. control and Ant-CB2 (n = 4). <b>B</b>: sp-TALP (control), Met-AEA (1.4 nM) or Met-AEA plus Ant-TRPV1 (TRPV1 antagonist capsazepine; 0.1 µM); **p<0.01 Met-AEA vs. control and Met-AEA plus Ant-TRPV1 (n = 6). <b>C</b>: sp-TALP (control), URB597 (FAAH inhibitor; 1 nM), URB597 plus Ant-CB1 or URB597 plus Ant-TRPV1; *p<0.05 URB597 vs. control, URB597 plus Ant-CB1 and URB597 plus Ant-TRPV1 (n = 5). (—) t = 0 and t = 45, sp-TALP at 0 and 45 min (control) incubation respectively.</p

Effect of micromolar concentration of Met-AEA on bull heparin-induced sperm capacitation.

<p>Spermatozoa were incubated for 45 min at 38.5°C in sp-TALP medium with heparin, Met-AEA (1.4 µM) or Heparin + Met-AEA (1.4 µM). Bars indicate the percentage of capacitated spermatozoa (A: % pattern B of CTC; B: % acrosome reacted spermatozoa). Data are expressed as mean ± SEM. <b>A</b>: Assessment of sperm capacitation by CTC assay. (—) t = 0 and t = 45, sp-TALP at 0 and 45 min (control) incubation respectively (n = 4). *p<0.05 vs control and Heparin (60 µg/ml) + Met-AEA (1.4 µM). <b>B</b>: Assessment of sperm capacitation by acrosome reaction evaluated by PSA-FITC; (—) t = 45 (control) (n = 4). ***p<0.001 vs control; **p<0.01 vs Heparin (60 µg/ml) + Met-AEA (1.4 µM).</p