1. The contractile effects of platelet activating factor (PAF) were compared in the myometrium isolated from non-pregnant and pregnant rats. 2. In the non-pregnant myometrium, PAF, at a concentration of 0.1 microM, did not change muscle tension and induced only a small transient contraction at 10 microM. 3. The contractile responses to PAF increased with the progress of gestation. In the late pregnant myometrium (21 day after gestation), PAF (0.1 nM-10 microM) induced large and relatively sustained contractions. The threshold concentration of PAF was decreased by approximately 10,000 times and the maximum contraction was increased 5 times by day 21 of gestation. 4. PAF (10 microM) increased the cytosolic Ca2+ concentration ([Ca2+] i) and muscle contraction to levels higher than those induced by high K+ in the pregnant rat myometrium (day 21). Verapamil (10 microM), a voltage-dependent Ca2+ channel blocker, decreased the stimulated [Ca2+] i and muscle tension to 49.6% and 22.7%, respectively, while the same concentration of verapamil completely inhibited the high K (+)-induced responses. 5. PAF (10 microM) induced a transient increase in [Ca2+] i with no contraction in the absence of external Ca2+ in the pregnant myometrium (day 21). 6. These results suggest that PAF induces contraction in rat myometrium by increasing Ca2+ influx. Although PAF released Ca2+ from stored sites, this Ca2+ does not seem to contribute to the PAF-induced contraction. Our major finding is that the sensitivity of the myometrium to PAF increased after gestation and that this may play a role in delivery.