The effects of motilin on gastrointestinal muscles show great variations in different organs and different species. For a precise regional differentiation, we recorded the mechanical activity of longitudinal and circular strips from fundus, corpus and antrum and of circular preparations from the inner and outer layer of the pyloric sphincter and from the duodenum (20 human stomachs). Motilin produced excitatory effects on the mechanical activity of the circular muscle strips from all regions of the human stomach including the pylorus. The effects on longitudinal preparations and on duodenal strips were weak. The most striking effect was an increase of phasic activity (amplitude) in circular antrum preparations, which exceeded the acetylcholine- and bombesin-induced activity. In pylorus preparations, a strong stimulation of phasic activity was observed with a transition to tonic activity in the inner layer of the pyloric ring at high motilin concentrations. The motilin-induced activity of the pyloric preparations was greater than the acetylcholine-induced contractions and even exceeded the bombesin-induced responses in the outer pylorus. The responses of the muscle strips of the proximal stomach (fundus and corpus) were weaker and did not exceed the acetylcholine-induced activity. All effects remained unaltered by atropine and tetrodotoxin application. The study confirms that motilin can interact directly with the smooth muscle of human stomach.