In order to elucidate systematically the effects of serotonin on gastric motility of guinea-pig, the contractile and electrical responses to serotonin were recorded using four kinds of muscle strips prepared from antral circular, antral longitudinal, fundic circular, and fundic longitudinal muscles. Experiments were performed using various methods including isometric contraction recording, transmural electrical field stimulation, junction potential recording, intracellular microelectrode technique, and partition stimulation method. The results were as follows: 1) The effect of serotonin on spontaneous contractions was inhibitory in the circular muscle strips of the antrum and fundus, while it was excitatory in the longitudinal muscle strips of the antrum and fundus. Serotonin changed mainly phasic contractions of both the circular and longitudinal muscle strips in the antrum, while it changed mainly tonic contractions of both the circular and longitudinal muscle strips in the fundus. 2) On the contractions induced by transmural nerve stimulation, serotonin decreased the amplitude in the circular muscle strips of the antrum, but it increased them in the other three groups of muscle strips(antral longitudinal, fundic circular, and fundic longitudinal). 3) On the contractions induced by direct muscle stimulation, serotonin decreased the amplitude in the circular muscle strips of the antrum and fundus. 4) In the fundic circular muscle strips serotonin potentiated excitatory junction potentials (EJPs), and in the antral circular muscle strips it evoked EJPs after inhibitory junction potentials(IJPS). 5) In the antral circular muscle strips serotonin did not affect the slow wave even at the disappearance of spontaneous contractions. On the contrary it increased the amplitude of the slow wave, when the spike component was potentiated and the second component was inhibited. 6) In the antral circular muscle strips the membrane potential was slightly hyperpolarized, but the membrane resistance was not changed. From the above results following conclusions could be made. 1) Serotonin inhibits spontaneous contractions of the circular muscle layer and it increases those of the longitudinal one, irrespective of the gastric region. 2) In the guinea-pig stomach there exists a serotoninergic facilitatory neuromodulation system which exerts its effect on cholinergically mediated contraction. 3) The excitation-contraction decoupling was observed in the effect of serotonin.