The potential involvement of cell-surface carbohydrates in sperm-egg fusion in mice was evaluated in this study. Zona-free mouse eggs were inseminated in the presence of a variety of simple saccharides to determine if certain sugars would act as competitive inhibitors of sperm-egg fusion. Of the sugars tested, L-fucose, galactose, and N-acetylglucosamine caused the greatest inhibition of sperm penetration levels relative to controls. A number of complex saccharides or glycoproteins with differing carbohydrate structures, including fucoidan, ascophyllan, ovomucoid, ovalbumin, fetuin, asialofetuin, and chondroitin sulfate, were also tested as competitive inhibitors of fusion. Only the L-fucose containing saccharides fucoidan and ascophyllan caused significant inhibition of fusion at concentrations of 0.05-1.0 mg/ml and 0.1-5.0 mg/ml, respectively. None of the other compounds tested had any inhibitory effect on fusion when tested at concentrations up to 5 mg/ml. The effect of the inhibitory saccharides was not due to the presence of residual zona material on the surface of the zona-free eggs, since zona-free eggs did not bind an 125I-labeled antibody directed against the ZP3 protein of the mouse zona pellucida. Pretreatment of either sperm or eggs with fucoidan (1 mg/ml) for 60 min prior to insemination caused only small decreases in sperm penetration levels, indicating that fucoidan exerted its major inhibitory effect on fusion only when present during insemination. Treatment of sperm, but not zona-free eggs, with fucosidase prior to insemination caused significant reductions in sperm penetration levels. Other glycosidic enzymes, including glucosidase, galactosidase, and N-acetylglucosaminidase, had no inhibitory effect on the sperm. These data suggest that an L-fucose component of the sperm surface is involved in sperm-egg fusion in the mouse.