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Analysis of Luteal Tissue Inhibitor of Metalloproteinase-1, -2, and -3 During Prostaglandin F₂-Induced Luteolysis¹

^a Department of Animal Science, University of Missouri, Columbia, Missouri 65211 ^b Departments of Animal Science and Physiology, Michigan State University, East Lansing, Michigan 48824

Increased matrix metalloproteinase (MMP) expression and activities help to mediate tissue involution through increasing extracellular matrix remodeling and promoting dedifferentiation and, ultimately, apoptosis. Therefore, we hypothesized that prostaglandin (PG) F₂ administration would decrease expression of the tissue inhibitor of metalloproteinase (TIMP)-1, -2, and -3 and effectively increase the MMP:TIMP ratio, leading to glandular involution. In experiment 1, we tested the effects of PGF₂ administration (Day 10 postestrus; Day 0 = estrus) on luteal TIMP-1, -2, and -3 mRNA and protein expression. Corpora lutea were collected at 0, 15, or 30 min or at 1, 2, 4, 6, 12, 24, and 48 h following PGF₂ administration (n = 3–9 animals/time point). Following PGF₂ administration, TIMP-1 mRNA levels decreased (P < 0.05) at 1 and 2 h relative to 0 h (controls), then increased to levels greater than controls at 4 and 6 h. In contrast, TIMP-2 and -3 mRNA levels did not decrease following PGF₂ administration. The TIMP-1, -2, and -3 proteins were localized to large luteal cells (LLCs) within control (untreated) tissues. However, histodepletion of TIMP-1 within LLCs was evident within 30 min (earliest time point collected) following PGF₂ injection and continued through 48 h. Luteal concentration of TIMP-1, as determined by RIA, was decreased (P < 0.05) by 15 min (earliest time point collected) following PGF₂ administration and remained low through 48 h. In contrast, TIMP-2 and -3 immunolocalization was not altered by PGF₂ administration. Experiment 2 was conducted to determine if PGF₂ could initiate the preceding changes in TIMP-1 in early (Day 3) corpora lutea that can bind PGF₂ but are refractory to its luteolytic effects. Serum concentrations of progesterone and luteal concentrations of TIMP-1 mRNA and protein were similar at 0 and 6 h after PGF₂ injection on Day 3 postestrus. These data suggest that an early and sustained effect of PGF₂ is the specific depletion of TIMP-1 within LLCs that are capable of responding to the luteolytic action of PGF₂. This action may increase the MMP:TIMP-1 ratio, creating an environment that favors extracellular matrix degradation and, thereby, facilitates both functional and structural regression.

First decision: 21 May 2001.

¹ Supported by U.S. Department of Agriculture grants 98-35203-6282 (M.F.S.) and 98-35203-6226 (G.W.S.). Contribution from the Missouri Agricultural Experiment Station, Journal Series No. 13, 107.

² Correspondence: Michael F. Smith, Department of Animal Sciences, 160 Animal Science Center, Columbia, MO 65211. FAX: 573 884 7827; smithmf{at}missouri.edu

³ Current address: Department of Anatomy, University of California at San Francisco, San Francisco, CA 94122

⁴ Current address: Bethyl Labs, P.O. Box 850, Montgomery, TX 77356

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