The luteolytic potency of fenprostalene (a PGF2alpha analog) is about 20-times that of naturally produced PGF2alpha. The objective of this research was to investigate the uterokinetic effects of fenprostalene at a luteolytic dosage (1.0 mg) in the cyclic and early postpartum cow, and in the isolated uterine horn. Uterine motility measurements were conducted on two consecutive days in each cow. Experimental protocol on Day 1 was: spontaneous motility was recorded for 1 h; fenprostalene was injected (1.0 mg i.m.), after which motility was recorded for 2 h; fenprostalene was injected (1.0 mg i.v.) and motility was recorded for 30 min; and oxytocin was injected (40 U i.v.), followed by a 30-min recording period. On Day 2, the treatment sequence was reversed: spontaneous motility was recorded for 1 h; oxytocin was injected (100 U i.m.), after which motility was recorded for 2 h; fenprostalene was injected (1.0 mg i.v.) and motility recorded for 30 min; and oxytocin was injected (40 U i.v.), followed by a 30-min recording period. In the in vitro experiment, different dosages of fenprostalene (5.9, 11.8, 17.6, and 29.4 ng/ml bath solutions) and oxytocin (0.06, 0.12, 0.18, and 0.60 mU/ml bath solutions) were tested in pairs for 1 h. The treatment was then repeated. In a different group, fenprostalene (5.9 ng/ml bath solution) and oxytocin (0.06 mU/ml bath solution) treatments were alternated. Fenprostalene (at luteolytic dosage) was not uterokinetic in either the cyclic or postpartum cow. However, fenprostalene and oxytocin had a significant uterokinetic effect (five- to six-fold pretreatment value) on the isolated uterine horn preparation at all dosages studied. Peak motility occurred between 10 to 15 min, followed by a gradual decrease to 40% at 60 min. When the treatments were repeated at 60 min, oxytocin but not fenprostalene caused a minute, transient contraction. However, fenprostalene-desensitized (by exposure to fenprostalene) uteri reacted significantly to oxytocin, and vice versa.