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Regular Article |
a Department of Animal Science, University of Missouri, Columbia, Missouri 65211
b Departments of Animal Sciences and Physiology, Michigan State University, East Lansing, Michigan 48824
c Department of Animal and Range Sciences and
d Cell Biology Center, Biotechnology Institute, North Dakota State University, Fargo, North Dakota 58105
Members of the matrix metalloproteinase (MMP) family collectively degrade extracellular matrix (ECM) and help regulate luteal function. The objectives of these experiments were to characterize the mRNA expression, localization, and activity of MMPs 2, 9, and 14 in ovine corpora lutea (CL). Ovine CL were collected on Days 2, 4, 10, and 15 of the estrous cycle (Day 0 = estrus). Messenger RNA transcripts for MMPs 2 and 14 were detected using Northern analysis; however, expression of MMP-9 was undetectable. Expression of MMP-14 mRNA (membrane type-1 MMP) was increased (P < 0.05) on Day 4; whereas, expression of MMP-2 mRNA was highest (P < 0.05) on Day 10, which corresponded to the observed increases in gelatinolytic activity in luteal homogenates as measured by a fluroscein-labeled gelatin substrate assay. MMP 2 and 9 proteins were localized predominantly to large luteal cells (LLCs), whereas MMP-14 was localized primarily to cells other than LLCs as demonstrated by immunohistochemistry. Immunolocalization of MMP-2 to putative endothelial cells was also observed on Day 15. Localization of MMP activity was determined using in situ zymography. Luteal tissues contained gelatinolytic activity primarily localized pericellularly to various cell types, including LLCs. These results support the hypothesis that ECM remodeling occurs throughout the luteal phase and may help potentiate cellular migration, differentiation, angiogenesis, and growth factor bioavailability.
1 This work was supported by USDA 98-35203-6282 (M.F.S.) and USDA 98-35203-6226 (G.W.S.) and is a contribution from the Missouri Agricultural Experiment Station Journal Series, no. 13151.
2 Correspondence: Michael F. Smith, Department of Animal Science, 160 Animal Sciences Center, University of Missouri, Columbia, MO 65211. FAX: 573 884 7827; smithmf{at}missouri.edu
3 Current address: Departments of Anatomy and Obstetrics and Gynecology, University of California, San Francisco, CA 94143
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