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Capacitated acrosome-intact mouse spermatozoa bind to the egg's zona pellucida in a receptor-ligand-mediated manner. Mouse zona pellucida 3 (mZP3) is a glycoprotein that functions as a primary ligand and inducer of the acrosome reaction (AR). Multiple sugar residues on mZP3 are thought to be recognized by complementary sugar binding enzymes (glycosidases or glycosyltransferases) or sugar binding lectin-like proteins on the sperm surface. To elucidate the nature of the sugar residues involved in sperm-egg recognition, several neoglycoproteins (ngps) were tested for their ability to induce the AR. Ngps are synthetic glycoproteins with a known monosaccharide conjugated to BSA. Capacitated mouse spermatozoa were treated in the absence or presence of several concentrations of ngps. A significantly greater number of spermatozoa underwent the AR in the presence of mannose-BSA, N-acetylglucosamine-BSA, and N-acetylgalactosamine-BSA than in their absence. Glucose-BSA or galactose-BSA had no effect on the AR. Inclusion of millimolar concentrations of unconjugated sugars (mannose, N-acetylglucosamine, or N-acetylgalactosamine) neither induced the AR nor blocked induction of the AR by ngps. These results demonstrate that some sugar residues can induce the AR, but only when conjugated to a protein backbone. Glucosaminyl-BSA (but not mannosyl-BSA or galactosaminyl-BSA) was a substrate for sperm-surface galactosyltransferase (GT), an enzyme thought to function as a receptor by binding to complementary glucosaminyl residues on mZP3. These data suggest a possible interaction between protein-conjugated glucosaminyl residues and sperm GT in the induction of the AR.

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Mammalian spermatozoa undergo the acrosome reaction (AR) in response to the interaction of a carbohydrate-recognizing molecule(s) on the sperm plasma membrane (sperm surface receptor) and its complementary glycan (ligand) moiety(ies) on the zona pellucida (ZP). Previously, we demonstrated that a hexose (mannose) or two amino sugars (glucosaminyl or galactosaminyl residues) when covalently conjugated to a protein backbone (neoglycoproteins) mimicked the mouse ZP3 glycoprotein and induced the AR in capacitated mouse spermatozoa (Loeser and Tulsiani, Biol Reprod 1999; 60:94-101). To elucidate the mechanism underlying sperm-neoglycoprotein interaction and the induction of the AR, we have examined the effect of several AR blockers on neoglycoprotein-induced AR. Our data demonstrate that two known L-type Ca(2+) channel blockers prevented the induction of the AR by three neoglycoproteins (mannose-BSA, N-acetylglucosamine-BSA, and N-acetylgalactosamine-BSA). The fact that the L-type Ca(2+) channel blockers (verapamil, diltiazem) had no inhibitory effect on sperm surface galactosyltransferase or alpha-D-mannosidase, two carbohydrate-recognizing enzymes thought to be sperm surface receptors, suggests that the reagents block the AR by a mechanism other than binding to the active site of the enzymes.