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Identification and Localization of Plasminogen Activator Inhibitor-1 Within the Porcine Oviduct¹

^a Departments of Animal Science, ^b Obstetrics and Gynecology, and ^c Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610-0294

The porcine oviduct synthesizes de novo and secretes a number of proteins into culture medium, many of which are unidentified. The objectives of the present study were to 1) semipurify and identify a M_r 45 000 secreted protein of the oviduct, 2) examine its synthesis within the three functional segments (infundibulum, ampulla, and isthmus), and 3) evaluate its distribution throughout the oviduct. Oviductal tissue was collected during early pregnancy, divided into functional segments, and subsequently cultured. Medium was collected, and the M_r 45 000 protein was concentrated by gel-filtration chromatography. The semipurified protein was transferred onto a polyvinylidene fluoride membrane and subjected to N-terminal amino acid analysis. The 26-amino acid sequence was 96% identical to that of pig plasminogen activator inhibitor (PAI)-1. Analysis by 1-dimensional SDS-PAGE and fluorography of rabbit anti-human PAI-1-immunoprecipitated product confirmed PAI-1. Subsequent 2-dimensional SDS-PAGE and fluorographic analyses of media revealed greater PAI-1 synthesis by the isthmus than by the ampulla or infundibulum. PAI-1 was immunolocalized throughout the oviduct and was heavily concentrated in the apical region of epithelial cells. Immunogold electron microscopy localized PAI-1 within putative secretory granules in the epithelial apical region and also associated with cilia in the isthmus. Isthmic PAI expression suggests a crucial role in protecting the preimplantation embryo from proteolytic degradation as well as in regulation of extracellular matrix turnover and remodeling.

First decision: 23 September 1999.

¹ Supported by USDA grant 95-37203-2308 and 97-35203-4614

² Correspondence: William C. Buhi, P.O. Box 100294, Department of Obstetrics and Gynecology, University of Florida, Gainesville, FL 32610-0294. FAX: 352-392-2808; buhiwc{at}obgyn.ufl.edu

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