Characterization of projections of longitudinal muscle motor neurons in human colon

Background The enteric nervous system contains inhibitory and excitatory motor neurons which modulate smooth muscle contractility. Cell bodies of longitudinal muscle motor neurons have not been identified in human intestine. Methods We used retrograde tracing ex vivo with DiI, with multiple labeling immunohistochemistry, to characterize motor neurons innervating tenial and inter‐tenial longitudinal muscle of human colon. Key Results The most abundant immunohistochemical markers in the tertiary plexus were vesicular acetylcholine transporter, nitric oxide synthase (NOS), and vasoactive intestinal polypeptide (VIP). Of retrogradely traced motor neurons innervating inter‐tenial longitudinal muscle, 95% were located within 6mm oral or anal to the DiI application site. Excitatory motor neuron cell bodies, immunoreactive for choline acetyltransferase (ChAT), were clustered aborally, whereas NOS‐immunoreactive cell bodies were distributed either side of the DiI application site. Motor neurons had small cell bodies, averaging 438 + 18µm2 in cross‐sectional area, similar for ChAT‐ and NOS‐immunoreactive subtypes. Motor neurons innervating the tenia had slightly longer axial projections, with 95% located within 9mm. ChAT‐immunoreactive excitatory motor neurons to tenia were clustered aborally, whereas NOS‐immunoreactive inhibitory motor neurons had both ascending and descending projections. VIP immunoreactivity was rarely present without NOS immunoreactivity in motor neurons. Conclusions and Inferences Tenial and inter‐tenial motor neurons innervating the longitudinal muscle have short projections. Inhibitory motor neurons have less polarized projections than cholinergic excitatory motor neurons. Longitudinal and circular muscle layers are innervated by distinct local populations of excitatory and inhibitory motor neurons. A population of human enteric neurons that contribute significantly to colonic motility has been characterized. Identification of longitudinal muscle motor neurons in human colon. Three longitudinal muscle motor neurons (arrows) in a single myenteric ganglion of human colon, retrogradely labelled by DiI (red) applied to the tertiary plexus. All 3 neurons are immunoreactive for choline acetyltransferase (green) but not nitric oxide synthase (blue) and hence likely to be excitatory motor neurons.