The present study determined the ovarian cellular localization of the mRNA for the tissue inhibitors of metalloproteinases (TIMPs) during pseudopregnancy in the rat. Pseudopregnancy was induced by eCG/hCG stimulation. At Day 1 of pseudopregnancy, intense reaction product for TIMP-1 mRNA was observed surrounding the developing corpus luteum (CL), with less intense expression present in granulosa-lutein cells. With continued luteal development, the TIMP-1 mRNA encircling the CL was lost, although low levels of expression were found within the CL. For TIMP-2 mRNA, intense reaction product was observed surrounding the developing CL but, unlike TIMP-1, was present in granulosa-lutein cells, with high levels near the center of the CL. The localization pattern of TIMP-2 mRNA was unchanged through the latter stages of pseudopregnancy. TIMP-3 mRNA expression was strikingly different from the other TIMPs. At Day 1 of pseudopregnancy, intense reaction product for TIMP-3 mRNA was observed in granulosa-lutein cells of certain developing CL, whereas adjacent follicles did not express TIMP-3 mRNA. With continued luteal development, there was a homogenous, intense localization of TIMP-3 mRNA throughout the CL, which was unchanged during pseudopregnancy. To understand the induction of TIMP-3 mRNA in the developing CL, a series of experiments was performed to compare markers of follicular maturity with the presence of TIMP-3 mRNA. TIMP-3 mRNA appears to be switched on in granulosa cells of follicles destined to ovulate. The distinct pattern of expression of the three TIMPs suggests that each inhibitor may regulate either the site and extent of proteolytic action or specific matrix metalloproteinases at different periods of the luteal life span.
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