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Matrix metalloproteinase‐2 (MMP‐2) expression and regulation by tumor necrosis factor alpha (TNFα) in the bovine corpus luteum
Author(s) -
Zhang Bo,
Yan Li,
Tsang Paul C.W.,
Moses Marsha A.
Publication year - 2005
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.20196
Subject(s) - biology , corpus luteum , angiogenesis , luteal phase , matrix metalloproteinase , estrous cycle , endocrinology , microbiology and biotechnology , medicine , northern blot , cdna library , gene expression , andrology , ovary , gene , cancer research , follicular phase , biochemistry
Angiogenesis and tissue remodeling events in the corpus luteum (CL) are mediated by matrix metalloproteinases (MMPs). We have recently reported the cloning of bovine membrane‐type 1 metalloproteinase (MT1‐MMP) and have shown that active MT1‐MMP is correlated to MMP‐2 activity in the CL during the estrous cycle. Given the important role that MMP‐2 plays in neovascularization, we became interested in understanding the role of this enzyme in the CL, a system in which angiogenesis is exquisitely regulated in the course of its lifespan. The aims of the present study were to clone bovine MMP‐2 cDNA, to investigate its temporal and spatial expression in three stages of CL during the estrous cycle and to study its regulation by TNFα, a key cytokine regulator of CL physiology. Bovine MMP‐2 cDNA was isolated from a UNI‐ZAP II bovine capillary endothelial cell cDNA library and sequenced. This gene encoded a protein of 662 amino acids. Luteal tissues were collected from non‐lactating dairy cows on days 4, 10, and 16 of the estrous cycle. Northern and Western blotting revealed that the levels of MMP‐2 mRNA (3.1 kb) and immunoreactive pro‐MMP‐2 protein (68 kDa) did not differ ( P > 0.05) in any age of CL examined. In addition to large luteal cells, MMP2 was localized to endothelial cells in all ages of CL by immunohistochemistry. Studies using in vitro luteal cell cultures showed that MMP‐2 mRNA, protein expression and activity was upregulated by TNFα in a dose‐ and time‐dependent manner. The present study suggests that MMP‐2 is predominantly produced by large luteal cells and endothelial cells, and that it plays an essential role in luteal remodeling and angiogenesis. These data also suggest that cytokines such as TNFα may modulate these processes by regulating MMP‐2 expression. Mol. Reprod. Dev. 70: 122–132, 2005. © 2005 Wiley‐Liss, Inc.