In this experimental study, we aimed to investigate the role of melatonin on uterine myoelectrical activity of non-pregnant rats. Forty-six female rats were assigned to six groups: (1) control; (0.2 mL 0.9% NaCl was injected intravenously (IV), n=6); (2) melatonin applied as 0.4 mg/kg/IV (n=8); (3) melatonin applied as 4 mg/kg/IV (n=8); (4) single dose of oxytocin (100 mU/kg) injected IV (n=8); (5) melatonin (0.4 mg/kg) plus oxytocin (100 mU/kg) (n=8); and (6) melatonin (4 mg/kg) plus oxytocin (100 mU/kg) injected IV (n=8). Each rat underwent a laparotomy, and uterine myoelectrical signals were recorded. The mean spectrum, averaged over the spectral content of signals in each group, was compared. Melatonin induced uterine myoelectrical activity in a dose-dependent manner. Treatment of melatonin after oxytocin suppressed the mean power of the signals. Serum melatonin concentrations were significantly higher in melatonin-treated rats. Melatonin itself at two different dose levels was found to be equally effective in stimulating the uterine electrical signals, although oxytocin-induced uterine electrical activity was suppressed by melatonin. These findings merit further investigations on the possible beneficial role of melatonin in the treatment of conditions associated with abnormal uterine activity.