Bovine membrane-type 1 matrix metalloproteinase: molecular cloning and expression in the corpus luteum.
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 ( approximately 63 kDa) and the active form ( approximately 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.
Zhang
Bo
B
Department of Animal and Nutritional Sciences, University of New Hampshire, Durham, NH 03824, USA.
Yan
Li
L
Moses
Marsha A
MA
Tsang
Paul C W
PC
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002417:Cattle; D003001:Cloning, Molecular; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D003720:Densitometry; D030762:Estrous Cycle / Q000502:physiology; D005260:Female; D005780:Gelatin / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D007150:Immunohistochemistry; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis* / Q000235:genetics; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014018:Tissue Distribution
D018076:DNA, Complementary; D012333:RNA, Messenger; D005780:Gelatin; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1