Figure 6
Role of voltage operated Ca2+ channels (VOCCs) in ZP1 273-551aa - mediated induction of acrosomal exocytosis. Capacitated sperm (1 × 106/100 μl) pre-treated with 10 μM Verapamil, 10 μM of Nifedipine (10 min prior-incubation with each to study effect of blocking of L-type VOCC), 10 μM Pimozide and 100 μM Amiloride (10 min pre-incubation; to study effect of blocking of T-type VOCC) were incubated with 2 μg/ml of ZP1273-551a for 60 min and subsequently analyzed for acrosomal status by TRITC-PSA staining as described in Methods. Spontaneous acrosome reaction (spontaneous AR) was analyzed by incubating sperm only with BWW medium. Additional negative controls comprised of the same amount of dimethyl sulphoxide (DMSO) and ethanol (EtOH) as used to dissolve T- and L-type VOCC inhibitors. 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 < 0.05, statistical significance with respect to the percent stimulation of acrosome reaction mediated by ZP1273-551aa.