In vitro effect of nickel on bovine spermatozoa motility and annexin V-labeled membrane changes

Abstract

In this study the effect of in vitro culture of bovine spermatozoa with nickel (NiCl(2)) on spermatozoa motility and membrane changes was analyzed. The spermatozoa motility significantly decreased after 120 min of culture at the concentration of 1000 μM Ni ml(-1) (P < 0.05) and after 240 min of culture at the concentration of 500 and 1000 μM Ni ml(-1) (P < 0.001) as compared with control. The progressive motility was the highest in the control group and in the groups with the lowest nickel concentrations (7.8 and 125 μM Ni ml(-1)). The progressive spermatozoa motility was significantly altered even after 30 min of culture in the group with the highest nickel concentration (1000 μM Ni ml(-1)). A significant decrease in progressive motility from the concentration of 250 μM Ni ml(-1) was detected after 240 min of culture. Concentrations from 125 μM Ni ml(-1) in various time periods of culture stimulated spermatozoa motility after 30 min (P < 0.001), but later an inhibitory effect was noted. After 240 min of in vitro spermatozoa culture with 125 μM Ni ml(-1) a typical Annexin V fluorescence reaction was detected. Fluorescence was detected in mitochondrial segment of bovine spermatozoa. In spermatozoa exposed to higher nickel concentrations the Annexin V-positive reaction was detected also on the spermatozoa head membrane. In the group with the highest concentration and the longest time of exposure (1000 μM Ni ml(-1); 240 min) the apoptotic Annexin-positive regions were detected not only in the mitochondrial part, but also in the spermatozoa head (acrosomal and postacrosomal part), showing significant alteration of spermatozoa membrane integrity.