PMID: 8957702Sep 1, 1996Paper

Anti-bull sperm monoclonal antibodies: II. Binding changes during capacitation and influence on sperm-zona interactions in vitro

Journal of Andrology
J D AmbroseC Y Lee

Abstract

Anti-bull sperm monoclonal antibodies (mAbs), generated against intra-acrosomal and surface antigens, were evaluated for their functional significance. In experiment I, the influence of mAbs on the bovine sperm-oocyte interaction in vitro was tested on a total of 493 oocytes in either three or four replicate trials. Although the number of sperm bound per zona increased significantly over untreated control samples (23.6 +/- 5.6 vs. 10.0 + 2.4, mean +/- standard error [SE]; P < 0.001) in the presence of one surface-reacting mAb, other mAbs had no effect. Experiment II was designed to determine if the mAbs would detect capacitation-related changes of bull sperm in vitro. Bull sperm were incubated in capacitation medium (supplemented Tyrode's medium [TALP] plus 10 micrograms/ml heparin) for up to 4.5 hours. At 0 and at 4 hours, mAbs in hybridoma culture supernatant were incubated for 30 minutes with sperm, labeled with fluorescein isothiocyanate (FITC)-conjugated secondary antibody, and processed for indirect immunofluorescence assay. Four mAbs specific to intra-acrosomal antigens exhibited a time-dependent increase (P < 0.05) in binding to bull sperm incubated under capacitation conditions. In contrast, the binding of the mAbs spe...Continue Reading

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