Effect of washing and capacitating media pH on bull sperm motility, acrosome integrity, and ability to penetrate zona-free hamster oocytes

Bovine-ejaculated sperm were washed thrice in bovine serum albumin-saline media, pH 7.2 to 8.4, and incubated at 37 degrees C in Ca++-free Tyrode's media. pH 7.2 to 8.4, for 0, 2, 4, 6, and 8 h. Motility was highest when sperm were washed in pH 7.2 medium and incubated in pH 8.0 or 8.4 media. Motility remained above 50% until 8 h. Washing in pH 7.6, 8.0, or 8.4 media induced more acrosome reactions after incubation than washing at pH 7.2. Percentage of acrosome-reacted sperm increased at each successive time period. Sperm penetrated more oocytes at 4, 6, and 8 h when wash medium pH was fixed at 7.2 and capacitating media pH was raised at .4 unit increments from 7.2 to 8.4. When sperm were washed in pH 7.2 medium, the postincubation penetration rates peaked at 8 h. With wash media of pH 7.6, 8.0, or 8.4, the postincubation penetration rates peaked at 4 h and then gradually declined. In conclusion, the most effective system for capacitating bull sperm was a pH 7.6 wash followed by capacitation in pH 7.6 medium for 4 to 8 h and this system resulted in the highest penetration rates. Wash media pH hastened capacitation but was not a capacitating agent.