Immunohistochemical techniques were used to examine the presence and co-localisation of a range of putative neurotransmitters and other neuronal markers in the myenteric plexus of the small and large intestine of the mouse. Distinct sub-populations of myenteric neurons were identified, based on the combinations of substances they contained and the distribution of their fibres. In the small intestine, there were two major classes of circular muscle motor neurons; one class was characterised by the presence of nitric oxide synthase, vasoactive intestinal peptide plus neuropeptide Y (NOS/VIP/NPY), and the second class contained calretinin plus substance P (CalR/SP). There were seven classes of neurons that innervated myenteric ganglia; these contained nos, vip, nos/vip, npy, calr/calbindin (calb), sp or 5-ht. In the large intestine, there were five major classes of motor neurons that contained nos, nos/vip, gaba, sp, or calr/sp, and seven major classes of neurons that innervated myenteric ganglia and contained nos, vip, calr/calb, calr, sp, gaba or 5-ht. Although some aspects of the patterns of co-localisation are similar to those in other species, this study re-inforces recent analyses that indicate significant species differences in neurochemical patterns in the enteric neurons of different species.

Background: The recent availability of antisera to the vesicular acetylcho-line transporter (VAChT) and choline acetyltransferase (ChAT) that demonstrate peripheral cholinergic neurons has made possible the anatomical identification of cholinergic neurons in the enteric nervous system. In this study, we localised cholinergic neurons in the mouse small and large intestine and identified which substances are found colocalised in the cholinergic neurons. Methods: Immunohistochemical single and double staining techniques were used on whole mount preparations and frozen sections to examine the localisation and chemical coding of cholinergic neurons in the small and large intestine of the mouse. Cholinergic neurons were identified using antisera to ChAT or VAChT. Results: In both the small and large intestine, numerous ChAT-immunoreactive nerve cell bodies were present in the myenteric and submu-cous ganglia, and ChAT-and VAChT-immunoreactive nerve terminals were abundant in the myenteric and submucous plexuses and the external muscle. Previous studies have identified two major classes of myenteric neurons in the small intestine of the mouse-those containing calretinin plus substance P, and those containing nitric oxide synthase (NOS) plus vasoactive intestinal peptide (VIP). Double-label studies showed that the vast majority of the calretinin/substance P neurons were cholinergic neurons, whereas only a small proportion of the NOS/VIP cells were cholinergic; the noncholinergic NOS/VIP neurons were motor neurons or interneurons, whereas the choliner-gic NOS/VIP neurons appeared to be exclusively interneurons. In the small intestine, all of the 5-HT-loaded neurons and a subpopulation of the calbindin neurons were also cholinergic. In the large intestine, there was a pattern of overlaps similar to that found in the small intestine, except that in the large intestine approximately 25% of the calretinin cells were not cholinergic. Only approximately one third of the GABA-loaded neurons in the large intestine were cholinergic. Conclusions: Large subpopulations of motor neurons and interneurons in the mouse small intestine are cholinergic neurons. Anat.

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The projections of different subpopulations of myenteric neurons in the mouse small and large intestine were examined by combining immunohistological techniques with myotomy and myectomy operations. The myotomies were used to examine the polarity of neurons projecting within the myenteric plexus and showed that neurons containing immunoreactivity for nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), calbindin and 5-HT projected anally, while neurons with substance P (SP)-immunoreactivity projected orally, in both the small and large intestine. Neurons containing neuropeptide Y (NPY)-and calretininimmunoreactivity projected locally. In the large intestine, GABA-immunoreactive neurons projected both orally and anally, with more axons tending to project anally. Myectomy operations revealed that circular muscle motor neurons containing NOS\VIP\pNPY and calretinin neurons projected anally both in the small and large intestine, while SP-immunoreactive circular muscle motor neurons projected orally. In the large intestine, GABA-IR circular muscle motor neurons projected both orally and anally. This study showed that although some neurons, such as the NOS\VP inhibitory motor neurons and interneurons, SP excitatory motor neurons and 5-HT interneurons had similar projections to those in other species, the projections of other chemical classes of neurons in the mouse intestine differed from those reported in other species.