Administration of estradiol-17beta (E) to ovariectomized (ovx) sheep results in the synthesis and release of an M(r) 90 000-92 000 glycoprotein into the oviductal lumen and into culture medium of ampullar explants (Biol Reprod 1992; 47:889-902). The objective of this study was to determine when and from what region of the oviduct the M(r) 90 000-92 000 glycoprotein is synthesized and released during early pregnancy. Estrous ewes were bred to intact rams of known fertility, and oviducts were obtained at estrus (Day 0) and at Days 1.5, 2, 3, 4, 6, and 16 of pregnancy. Pregnancy was verified by the presence of a fertilized egg or developing conceptus and a functional corpus luteum. Oviductal secretions were collected by flushing oviducts with saline and by explant culture. The oviductal fimbria, ampulla, and isthmus were individually cultured (24 h) in the presence of H-3-leucine (H-3-leu) or H-3-glucosamine (H-3-glcN). The presence of the M(r) 90 000-92 000 glycoprotein in oviductal flushing and culture medium was determined by fluorography and Western blotting. The M(r) 90 000-92 000 protein was present in SDS gels and blots of oviductal flushing from animals through Days 4-6 of pregnancy, but not in flushing from Day 16 pregnant animals or from ovx, untreated animals. This protein was present in H-3-leu- and H-3-glcN-labeled culture medium of the oviductal ampulla (Days 0, 1. 5, 2, 3, 4, 6, and 16) and fimbria (Days 0, 1. 5, 2, 3, and 4) during early pregnancy. The M(r) 90 000-92 000 protein was not detected in oviductal ampulla or fimbria culture medium obtained from ovx animals, nor was it present in culture medium of the isthmus oviduct obtained from estrous and pregnant animals. These data show that the E-dependent M(r) 90 000-92 000 glycoprotein is synthesized and released by the oviduct in a temporally and regionally specific manner during early pregnancy as the gametes and fertilized egg are traversing the tube and prior to a systemic rise in progesterone.