Figure 5
Role of extracellular calcium and G i protein-coupled receptor in ZP1 273-551aa - mediated induction of acrosomal exocytosis. Capacitated sperm (1 × 106/100 μl) were pre-treated with 8 mM EGTA (10 min prior-incubation) and 0.1 μg/ml PTX (30 min prior-incubation) to study effects of extracellular calcium and Gi protein respectively. These pretreated sperm were incubated with ZP1273-551aa (2 μg/ml) for 60 min, and subsequently analyzed for acrosomal status by TRITC-PSA staining as described in Methods. Spontaneous acrosome reaction was assessed by incubating sperm only with BWW medium. Data has been represented as percent stimulation of acrosome reaction calculated as described in Methods. Values are Mean ± SEM of 3 different experiments using semen samples from at least two different male donors. *p < 0.01, statistical significance with respect to the percent stimulation of acrosome reaction mediated by ZP1273-551aa.