Analysis of transcript abundance of uterine genes after application of bovine pregnancy associated glycoproteins to endometrial explants
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Pregnancy-associated glycoproteins (PAGs) belong to the aspartic protease family and are expressed exclusively by trophoblasts of even-toed ungulates. This study was designed to provide insight into some of the biological roles of bovine PAGs in ruminant pregnancy by measuring changes in transcript abundance in cultured endometrial explants from pregnant and non-pregnant heifers treated with PAGs. A mixture of PAG-4, -6, and -9 were purified from mid-gestation bovine placental extracts. Heifers were synchronized and bred by artificial insemination to either high fertility semen (n = 14) or dead semen (n = 6), then slaughtered at day 18 post-breeding. Endometrial explants were collected and split between 4 groups: pregnant + 15 μg/ml PAG (n = 10), pregnant + no PAG (n = 10), non-pregnant + 15 μg/ml PAG (n = 10), and non-pregnant + no PAG (n = 10). The transcript abundance for several tissue remodeling related target genes [extracellular matrix metalloproteinase inducer (EMMPRIN), matrix metalloproteinases (MMP) 1, 2, 3, 7, 8, and 9, plaminogen activator urokinase (PLAU), osteopontin (SPP1), tissue inhibitors of MMP 1 and 2 (TIMP1, TIMP2)], were analyzed in endometrial tissue by quantitative PCR. We observed consistent changes in transcript abundance between pregnancy groups for MMP1, 3, 7, 8, PLAU, SPP1 at 24 hours and MMP1, 3, 8, PLAU, SPP1 at 96 hours. These results indicate that the PAGs used in this experiment are capable of inducing changes in transcripts associated with matrix remodeling in bovine endometrial explants. This model system may be useful for assessing PAG function in the uterine stroma.
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