The ultrastructural localization of an oviductal glycoprotein, designated ZP-0 in golden hamster oviductal eggs, was investigated by immunolabeling methods using a monoclonal antibody (C11E8). Immunofluorescence staining showed that C11E8 specifically reacted with the zona pellucida of the oviductal egg but not the ovarian egg. In an immunoelectron microscopic study applying the protein-A gold technique, gold particles were distributed throughout the zona pellucida of the oviductal eggs and were also associated with the perivitelline matrix. Structures within the eggs and cumulus cells did not react with C11E8. Quantitative evaluations of the degree of labeling demonstrated that a large number of gold particles was bound to the zone pellucida, especially in the middle layer. Moreover, in bovine testicular hyaluronidase-treated eggs the density of labeling decreased only in the outer third of the zona pellucida. These results show that ZP-0 to the was associated with the zona pellucida and perivitelline matrix of the golden hamster egg after ovulation and suggest that there are topographical differences in the binding activity of ZP-0 to the zona pellucida. In addition, the decrease in labeling density of ZP-O induced by hyaluronidase appears to be related to changes in the properties of the outer layer of the zona pellucida.