To investigate expression of monocyte chemoattractant protein-1 (MCP-1) in the ovine corpus luteum, a partial cDNA was produced by reverse transcription-polymerase chain reaction. This cDNA was 89% identical to that reported for bovine MCP-1 mRNA. In experiment 1, steady-state concentrations of mRNA encoding MCP-1 were measured in pools of luteal tissue collected on Days 3, 6, 9, 12, and 15 of the estrous cycle (estrus = O; n = 4/day). There were no differences in mRNA concentrations for MCP-1 among any of the days studied (p = 0.43). In experiment 2, midluteal-phase corpora lutea were collected from ewes at 0 (untreated), 2, 4, 8, and 16 h after administration of a luteolytic dose of prostaglandin F2alpha (PGF2alpha; n = 4/time point). Concentrations of MCP-1 mRNA were undetectable in untreated controls, were detectable at 2 h post-treatment, had increased 4 and 8 h after administration of PGF2alpha when compared to those at 2 h (p < 0.05), and were decreased 16 h after administration of PGF2alpha when compared to those at 4 h (p < 0.05). In situ hybridization for MCP-1 mRNA combined with immunocytochemical labeling of tissue inhibitor of metalloproteinase-1 (TIMP-1) in large luteal cells was used to determine whether the steroidogenic cells that have PGF2alpha receptors express MCP-1 mRNA in response to PGF2alpha. Messenger RNA encoding MCP-1 and TIMP-1 were not colocalized, indicating that MCP-1 was not expressed by large steroidogenic luteal cells during luteolysis.