The effects of relaxin on contractility and membrane potential of the longitudinal and circular smooth muscle layers of the uterus have been studied in vitro using oestrogen‐treated, non‐pregnant rats and pregnant rats. Relaxin decreased the amplitude of contractions induced by electrical stimulation of longitudinal myometrium by decreasing the duration of the bursts of action potentials. This effect was transient and tachyphylaxis always developed and was observed following injection of steroids and up to day 17 of pregnancy. There was no inhibition of tissues from rats from day 18 of pregnancy to term. The peptide had no effect on resting membrane potential, space constant or time constant. Action potentials recorded from circular myometrium of non‐pregnant rats pre‐treated with oestrogen consisted of an initial spike or short burst of spikes followed by a prolonged plateau of depolarization. Spontaneous action potentials and associated contractions were abolished within 2 min of exposure to relaxin (10(‐8) g/ml) while contractions of much smaller amplitude could be evoked with depolarizing current pulses. This effect was associated with depression of the plateau component of the action potential whereas the spike component was left intact. Relaxin had no effect on passive membrane properties. The action potentials of circular myometrium of rats up to day 21 of pregnancy were qualitatively similar to those recorded in the same muscle layer from oestrogen‐treated, non‐pregnant rats and the plateau component was also blocked by relaxin in these tissues. Bursts of spikes were observed in circular strips 24‐36 h before parturition, and the effect of the peptide on these was a transient inhibition similar to that observed in longitudinal myometrium. Oxytocin increased the amplitude of the spike and the amplitude and duration of the plateau. Relaxin abolished the plateau in the presence of 10(‐11) and 10(‐10) M‐oxytocin but was ineffective when the concentration of the spasmogen was increased further. Prostaglandin F2 alpha increased the amplitude and duration of the plateau. Relaxin abolished the responses to 10(‐10) and 10(‐9) M‐prostaglandin F2 alpha. The results of this study demonstrate that relaxin specifically inhibits contractions in the circular layer of the myometrium by abolishing the plateau component of the action potential. This action appears to be different from that of other smooth muscle relaxants tested in these experiments (isoprenaline, papaverine and verapamil). All of these abolished simultaneously both the spike and plateau components of the action potential.