A preparation of isolated small intestine from the guinea-pig was studied in which reflex responses of the circular muscle were recorded intracellularly when sensory receptors in the mucosa were stimulated mechanically. This preparation was used to examine the properties of mucosa to muscle reflexes that involve non-cholinergic motor neurons innervating the circular muscle. Reproducible stimulation of the mucosa was achieved by stroking with a motor-driven brush. Gentle brush-strokes applied to the mucosa typically evoked inhibitory junction potentials anal to the stimulus and excitatory junction potentials at recording sites oral to the stimulus. Both events were rapid in onset and up to 25 mV in amplitude. The reflexes were blocked by tetrodotoxin (0.5 microM). Junction potentials declined in amplitude with distance from the stimulus, the amplitude of the excitation 15 mm oral to the stimulus was half that at 5 mm from the stimulus, whereas the amplitude of the inhibitory potential at 40-45 mm was about 60% of that at 5-10 mm anal to the stimulus. Hexamethonium (100-200 microM) blocked the ascending excitation but only slightly reduced the descending inhibition. Ascending excitation was blocked by antagonists for substance P receptors in the muscle, and inhibition was substantially reduced by apamin (0.2 microM), both before and after hexamethonium. Both responses were abolished by removal of the mucosa from the stimulus site and when lesions were made through the myenteric plexus between the stimulation and recording sites, but persisted when similar lesions were made through the submucous plexus. It is concluded that there are neurons with mechanoreceptive nerve endings in the mucosa. Stimulation of such sensory neurons leads to activation of pathways in the myenteric plexus that excite motor neurons to the muscle both oral and anal to the stimulation site. The demonstration that mucosa to muscle reflexes can be consistently evoked in the small intestine in vitro provides an opportunity for close analysis of the reflex pathways. Such analysis is not so readily achieved when reflexes are initiated by distension that, by moving the intestine, can dislodge the recording electrode.