We examined whether spontaneous parturition in sheep was associated with tissue-specific changes in prostaglandin H₂ synthase-2 (PGHS-2) expression and/or with altered expression of myometrial EP and FP receptors. Placental and uterine tissues were collected from three groups of chronically catheterized sheep in relation to term spontaneous labor: late pregnancy, not in labor; early labor; and active labor. Expression of PGHS-2 mRNA and protein was determined by in situ hybridization, Western blotting, and immunohistochemistry. Semiquantitative reverse transcription-polymerase chain reaction was used to assess the presence of and changes in prostaglandin (PG) receptor subtypes. In placenta, PGHS-2 mRNA and protein localized to trophoblast uninucleate cells and tended to increase with early labor. PGHS-2 mRNA and protein localized to endometrial epithelium and to myometrium, where PGHS-2 protein levels rose in active labor tissues. Concentrations of PGE₂ in fetal plasma rose progressively with labor, whereas 13,14-dihydro-15-keto-PGF_2α in maternal plasma increased significantly only in active labor. Messenger RNA encoding four EP receptor subtypes and FP receptor were present in myometrium, but levels did not change with labor. We suggest that spontaneous labor in sheep is associated with a progressive increase in PGHS-2 expression in a temporal and tissue-specific manner from trophoblast to maternal tissues, rather than alteration in PG receptor gene expression.