Quantitative estimation and chemical coding of spiny type I neurons in human intestines

Previous studies have shown that most human myenteric neurons co-staining for vasoactive intestinal peptide (VIP), neuronal nitric oxide synthase (nNOS) and neurofilaments (NF) display the morphology of spiny type I neurons displaying a descending projection pattern. Here, we estimated the proportions of spiny neurons in human intestines, the amount of congruence of VIP/nNOS-immunoreactive with spiny neurons and whether galanin (GAL) is co-localized with VIP. Three sets of colchicine-pretreated and fixed whole mounts of 21 patients or body donors (median age 65 years; 10 female, 11 male) were stained for VIP, nNOS and NF, for VIP, nNOS and the human neuronal protein Hu C/D (HU) as well as for VIP, nNOS and GAL. The majority of VIP/nNOS-co-reactive neurons were spiny neurons (79/80% in small/large intestine, respectively) and the majority of spiny neurons co-stained for VIP and nNOS (82/69%). Neurons co-immunoreactive for VIP/nNOS/HU amounted to 7 and 4%, respectively. GAL/VIP-co-immunoreactivity was demonstrated in 69 and 27% of spiny neurons, respectively. We conclude that the number of neurons displaying co-reactivity for VIP and nNOS is a quantitative indicator of spiny neurons in both small and large intestine and that the proportion of spiny neurons is about 7% in small and 4% in large intestines. Since nerve fibres co-staining for NF/VIP/nNOS were found mainly in the circular muscle layer but not the surrounding perikarya of spiny neurons, we suggest that they may represent inhibitory motor neurons rather than descending interneurons.