The zona pellucida binds the mature form of an oviductal glycoprotein (oviductin).
The use of a monoclonal antibody (MAb) specific for the oviductal zona pellucida (ZP) of the hamster has demonstrated that a new antigen (oviductin) is acquired by the ZP during transit of the oocyte in the oviduct. The epitope that is recognized by the MAb bears a terminal N-acetyl-D-galactosamine residue. We conducted a study in order to determine whether this immunoreactivity of the oviductal ZP results from the addition of the terminal sugar residue to a preformed ZP protein or from the transfer of the mature glycoprotein produced by oviductal secretory cells. We measured the incorporation of [35S]methionine into proteins using four different incubation systems: cumulus oophorus (CO) alone, CO in the presence of oviductal fluid, CO co-incubated with empty oviducts, and CO within intact oviducts. At the end of the incubation period, the ZP, vitelli, dispersed cumulus without oocyte, oviducts, and culture medium were isolated and analyzed for their protein content by sodiumdodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), autoradiography, and immunodetection. The cumulus cells synthesized several proteins, independently of the oviductal environment; however, none of these proteins corresponded to oviductin. The ZP and the vitelli of cumulus oophorus that were incubated either alone or in the presence of oviductal fluid did not contain radioactive oviductin. When the oviduct (empty or intact) was present in the incubation system, radiolabeled oviductin was synthesized and secreted into the incubation medium. The ZP picked up a detectable amount of radioactive antigen only in the system in which intact oviducts were incubated.(ABSTRACT TRUNCATED AT 250 WORDS)
St-Jacques
S
S
Department of Immunology, University of Montreal, Quebec, Canada.
Malette
B
B
Chevalier
S
S
Roberts
K D
KD
Bleau
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D006224:Cricetinae; D000939:Epitopes / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D008647:Mesocricetus; D010057:Oviducts / Q000378:metabolism*; D015044:Zona Pellucida / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins