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Responses of myenteric AH and S neurons to local application of chemicals to the mucosa of the guinea pig small intestine were obtained using conventional intracellular recording techniques. Preparations were dissected to reveal the myenteric plexus over one-half of the circumference of the gut with intact mucosa on the the half. Neurons were impaled within the exposed one-half, whereas potential stimulants, in buffered saline, were transiently applied to the mucosa, 1-1.5 mm circumferential from the impalement. The stimulants elicited action potentials (AP) in AH neurons that did not arise from synaptic activity. AH neurons also responded with slow excitatory postsynaptic potentials (EPSP). S neurons were activated synaptically, via fast and slow EPSP, but not nonsynaptically. Mucosal application of solutions of a low pH (3-5) or a high pH (9-11) were both effective stimulants. Solutions of a neutral pH, which was also a control for mild mechanical stimulation, were usually ineffective. Both a short-chain fatty acid, acetate (pH 7.2), and 5-hydroxytryptamine elicited responses in each neuronal type. We conclude that myenteric AH neurons of the guinea pig distal ileum are primary afferent neurons that respond to a variety of mucosally applied chemical stimuli with burst of AP. In addition, the physiologically evoked transmission of slow EPSP to AH neurons suggests that primary afferent neurons interconnect in a self-reinforcing network. S neurons are second or higher order neurons in the reflex pathways.