Secretion of proteins by cultured bovine oviducts collected from estrus through early diestrus.
To characterize in vitro protein secretion by the oviduct throughout early stages of the estrous cycle, 16 cows received a luteolytic dose of PGF2 alpha and were randomly assigned to be killed on day (D) 0, 2, 5, or 8 after subsequent estrus. Explant cultures of oviducts (ampulla and isthmus) were incubated for 24 h at 39 degrees C in a modified Eagle's minimal essential medium supplemented with 50 microCi L-[4,5-3H]leucine. Oviductal secretion of de novo synthesized protein measured by incorporation of [3H]leucine into nondialyzable radioactivity in culture supernatants was greatest at D 0 and declined thereafter. Incorporation of [3H]leucine into TCA-precipitable macromolecules in tissue homogenates was also greatest at D 0. Analysis of culture supernatants by one-dimensional polyacrylamide gel electrophoresis revealed four major bands of radiolabeled proteins at greater than 97 kDa, 85-97 kDa, 55 kDa, and 30 kDa. Analysis of individual polypeptides resolved by two-dimensional polyacrylamide gel electrophoresis indicated that five of 32 individual polypeptides examined were secreted at significantly greater rates at estrus than at other times examined. One of these five polypeptides, a 97-kDa peptide with an apparent pI of 5.0, was the major secretory product at estrus and accounted for 18% of total radioactivity recovered from two-dimensional gels. Two of 32 polypeptides examined were secreted at significantly greater rates by explants of the oviduct contralateral to the side of ovulation. In summary, estrus is associated with an elevation in total protein secretion by the bovine oviduct. This increase is due to selective amplification of secretion of several but not all secretory proteins.
Malayer
J R
JR
Dairy Science Department, University of Florida, Gainesville 32611.
Hansen
P J
PJ
Buhi
W C
WC
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004971:Estrus / Q000378:metabolism*; D005260:Female; D009924:Organ Culture Techniques; D010057:Oviducts / Q000378:metabolism*; D010779:Photofluorography; D014176:Protein Biosynthesis