1 The longitudinal muscle from the uterus of oestrogen-treated rats was quiescent in Mg-free Krebs solution. Electrical stimulation generated phasic contraction, which was depressed to 35% and 18% by 50 mu and 150 mu porcine relaxin, respectively. 2 The phasic contractions were more strongly depressed to 26% by 50 mu relaxin in solution containing 0.6 mm Mg, and the depression lasted for more than 4 h after the removal of relaxin. During the persisting depression, raising the external Ca to 7.5 mm did not restore the contraction, but the contraction was restored by removal of Mg. 3 The depression of the phasic contraction by relaxin, examined in Mg-free solution, was enhanced and reduced by pretreatment of the tissue with 0.6 mm Mg and 0.6 mm Mn, respectively, for about 15min. In contrast, the depression of contraction by isoprenaline or forskolin was enhanced by pretreatment with either Mg or Mn. 4 The cellular content of cyclic AMP was measured in Krebs solution containing 0.6 mm Mg. The values were 1.24 (pmolmg-' protein) in control solution, and 2.31 and 1.56 when the tissues were treated with 150 mu relaxin and 10-9 M isoprenaline, respectively. 5 The cyclic AMP production in response to 10-7M forskolin measured in Mg-free solution was enhanced when the tissue was pretreated with either 0.6mm Mg or Mn for 15min. The cyclic AMP production in response to 100mu relaxin was increased when the tissue was pretreated with 0.6mm Mg, and was unchanged by pretreatment with Mn. The cyclic AMP production in response to 10-9M isoprenaline was unchanged by pretreatment with the divalent cations. 6 The membrane potential of the muscle was -60.8mV in Krebs solution containing 0.3mm Mg, and electrical stimulation induced an action potential which consisted of spike and plateau components. Application of 150mu relaxin reduced the duration of the plateau; the contractions were progressively depressed. The resting membrane potential and membrane resistance were unchanged by application of 150 mu relaxin. The membrane was hyperpolarized by 2.8 mV, accompanied by a decrease in membrane resistance, when I0-9 M isoprenaline was applied. 7 Although there were several differences between the effects of relaxin and isoprenaline, it is probable that some process, which is cyclic AMP-dependent, accelerated by Mg and depressed by Mn, is involved in the depressant action of relaxin on contraction.