1 Whole cell patch clamp techniques were used on thin brainstem slices to investigate the effects of 5-hydroxytryptamine (5-HT) on gastrointestinal-projecting dorsal motor nucleus of the vagus (DMV) neurones. Neurones were identified as projecting to the stomach (n=122) or intestine (n=84) if they contained the fluorescent tracer Dil after it had been applied to the gastric fundus, corpus or antrum/pylorus or to the duodenum or caecum. 2 A higher proportion of intestinal neurones (69%) than gastric neurones (47%) responded to 5-HT with a concentration-dependent inward current which was antagonized fully by the 5-HT2A receptor antagonist ketanserin (1 microM). 3 Stimulation of the nucleus tractus solitarius (NTS) induced inhibitory synaptic currents that were reduced in amplitude by application of the 5-HT1A receptor agonist 8-OHDPAT (1 microM) or the 5-HT1A/1B receptor agonist TFMPP (1 microM) in 61% and 52% of gastric- and intestinal-projecting neurones, respectively. 5-HT also significantly reduced the frequency but not the amplitude of spontaneous inhibitory currents. 4 These data show that 5-HT excites directly a larger proportion of intestinal projecting neurones than gastric-projecting neurones, as well as inhibiting synaptic transmission from the NTS to the DMV. These data imply that the response to DMV neurones to 5-HT may be determined and classified by their specific projections.