The localization of reduced nicotinamide adenine dinucleotide phosphate diaphorase in the submucous plexus of duodenum, jejunum, ileum, proximal colon, distal colon and rectum in the guinea-pig was examined histochemically by light and electron microscopy. The majority of reactive submucous neurons displayed features common to either Dogiel type I or type II neurons; some were closely adherent to the outer walls of lymphatic vessels. The use of 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT) at the ultrastructural level showed that nicotinamide adenine dinucleotide phosphate diaphorase is a membrane-associated protein widely distributed in the cells, including the rough endoplasmic reticulum, Golgi apparatus and synaptic vesicles in the axon terminals associated with submucous neurons. On the basis of their diaphorase reactivity or the lack of it, the submucous neuronal somata and their associated terminals were observed to form several different kinds of synaptic configurations. The present quantitative analysis showed that the frequency of reactive submucous neurons in the large intestine was significantly higher than in the small intestine. Based on the ultrastructural localization of the diaphorase reaction product in positive cells, it is speculated that nitric oxide might be synthesized within the neurons. The demonstration of different synaptic configurations in the submucous ganglia suggests that the functional interaction between submucous neurons is extremely complex. Finally, the higher frequency of diaphorase reactive submucous neurons in the large intestine than in the small intestine indicates that submucous neurons in these two gut regions may not play equivalent roles.