Identification of matrix metalloproteinases and metalloproteinase inhibitors in bovine corpora lutea and their variation during the estrous cycle.

Corpora lutea (CL) were collected from cattle to study key physiologic events in angiogenesis. Our objective was to evaluate the activity of matrix metalloproteinases (MMP) and endogenous inhibitors. Corpora lutea were collected 2, 4, 6, 8, 10, 12, 14, and 16 d (n = 3/d) after estrus was first detected. In zymograms, a band of protein migrating at a relative molecular mass (M(r)) of 98 kDa was increased early in the cycle; a M(r) = 88 kDa band was detectable on all days. The molecular weights of these proteins are consistent with the MMP-9 family members. In all samples, a band of enzyme activity was detected at M(r) = 62 kDa, and another band of lesser density was detected at M(r) = 60 kDa. The molecular weights of these proteins are consistent with the MMP-2 family members. An immunoreactive band, detected in all samples with equal density, migrated between M(r) = 27 and 29 kDa, as did the tissue inhibitor of metalloproteinase (TIMP-1) standard. A second band, which was less dense in samples from d 2 through 6, migrated at M(r) = 19 kDa, as did the TIMP-2 standard. A third band was detected in all samples; it migrated at M(r) = 35 kDa, as did the cartilage-derived inhibitor (CDI) standard, and was less dense in d 8 and d 12 through 16 samples. In summary, MMP (gelatinases) and MMP inhibitors are present in developing luteal tissue, and the M(r) = 98 kDa enzyme, CDI, and TIMP-2 varied during the estrous cycle.