Abstract
Bovine sperm heads were separated via ultrasonic treatment and centrifugation. Anti‐bull sperm IgG was produced by immunizing rabbits with acrosome‐reacted bull sperm heads. SDS PAGE patterns revealed that the main membrane proteins on acrosome‐reacted bull sperm head were sp18 family, including 18, 16, and 14 kD, which represented about 64% of the total membrane proteins in bull sperm. Indirect immunofluorescence shown sp18 antigens primarily distributed in postacrosomal and proximal tail regions. Western blot analysis revealed that the anti‐bull sperm IgG reacted with sp18 antigens in acrosome‐reacted bull sperm head and bull seminal plasma. Anti‐bull sperm IgG also reacted with 14, 16, 18, 42, 57 and 60 kD proteins in fresh bull, mouse and rabbit sperm. Anti‐sp18 IgG caused agglutination of bull and rabbit sperm, but had no effect on murine sperm. In murine in vitro fertilization trials, preincubating capacitated sperm with 0.364 mg/ml of anti‐sp18 IgG resulted in a decrease in the fertilization rate from 75.6% in the controls to 50.8% in the experimental groups (p<0.001).
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