Comparison of Various Bovine Sperm Capacitation Systems for Their Ability to Alter the Net Negative Surface Charge of Spermatozoa

Abstract

Semen was divided into five aliquots and capacitated for 4 or 6h in calcium-free Tyrode's( 37°C), modified Tyrode's bovine follicular fluid (39°C), modified Tyrode's heparin (39°C), and TEST-yolk buffer (4°C) or incubated in BSA-saline (39°C). Sperm before and after capacitation were electrophoresed at 100V of constant current for 3 and 6min. Samples were collected, sperm were counted, and percentages of sperm migrating toward the anode at 0h and 4 or 6h were calculated. At 0h, 61 to 80% of the sperm migrated with a net negative charge. After 4 or 6h, the percentage of sperm migrating toward the anode significantly decreased in all capacitation media but not in the control.The percentage of sperm migrating toward anode was lowest in calcium-free Tyrode's (22.1%) followed by TEST (28.4%), Tyrode's follicular fluid (29.5%), and Tyrode's heparin (42.5%); these values were less than those for the control (56.0%). The movement of live sperm without the influence of current or freeze-killed sperm under the influence of electrical current (control trials) was negligible. Capacitation reduced the net negative surface charge of sperm, the magnitude of which depended on capacitation system. Net negative charge of sperm was inversely associated with efficiency in penetration of zona-free hamster oocytes.