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Bovine Membrane-Type 1 Matrix Metalloproteinase: Molecular Cloning and Expression in the Corpus Luteum¹

^a Department of Animal and Nutritional Sciences, University of New Hampshire, Durham, New Hampshire 03824 ^b Laboratory for Surgical Research, Department of Surgery, Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Matrix metalloproteinase-2 (MMP-2) is produced as a zymogen, which is subsequently activated by membrane-type 1 metalloproteinase (MT1-MMP). The objectives of the present study were to clone bovine MT1-MMP and to investigate its expression in the corpus luteum. Corpora lutea were harvested from nonlactating dairy cows on Days 4, 10, and 16 of the estrous cycle (Day 0 = estrus; n = 3 for each age). The bovine MT1-MMP cDNA contained an open reading frame of 1749 base pairs, which encoded a predicted protein of 582 amino acids. Northern blotting revealed no differences (P > 0.05) in MT1-MMP mRNA levels between any ages of corpora lutea. Western blotting demonstrated that two species of MT1-MMP, the latent form (63 kDa) and the active form (60 kDa), were present in corpora lutea throughout the estrous cycle. Active MT1-MMP was lower (P < 0.05) in early stages of the corpus luteum than the mid and late stages, where MMP-2 activity, as revealed by gelatin zymography, was also elevated. Furthermore, immunohistochemistry revealed that MT1-MMP was localized in endothelial, large luteal, and fibroblast cells of the corpus luteum at different stages. Taken together, the differential expression and localization of MT1-MMP in the corpus luteum suggest that it may have multiple functions throughout the course of the estrous cycle, including activation of pro-MMP-2.

First decision: 3 October 2001.

¹ This work was supported by a research grant from American Cancer Society (ACS-RPG-97-013-CB) to M.A.M., the Northeast Regional Project NE-161 to P.C.W.T., and the U.S. Department of Agriculture (98-35208-6654). This work was presented in part at the 33rd Annual Meeting of the Society of the Study for Reproduction at Madison, Wisconsin. This is scientific contribution 2104 from the New Hampshire Agricultural Experiment Station.

² Correspondence. FAX: 603 862 3758; pct{at}cisunix.unh.edu

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