de Vries P, Soret R, Suply E, Heloury Y, Neunlist M. Postnatal development of myenteric neurochemical phenotype and impact on neuromuscular transmission in the rat colon. Am J Physiol Gastrointest Liver Physiol 299: G539 -G547, 2010. First published June 3, 2010 doi:10.1152/ajpgi.00092.2010.-Profound changes in intestinal motility occur during the postnatal period, but the involvement of the enteric nervous system (ENS), a key regulator of gastrointestinal (GI) motility, in these modifications remains largely unknown. We therefore investigated the postnatal development of the ENS phenotype and determined its functional repercussion on the neuromuscular transmission in the rat colon. Sprague-Dawley rats were euthanized at postnatal day (P) 1, P3, P5, P7, P14, P21, and P36. Whole mounts of colonic myenteric plexus were stained with antibodies against choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), and HuC/D. Colonic contractile response induced by electrical field stimulation (EFS) was investigated in organ chambers in absence or presence of N-nitro-L-arginine methyl ester (L-NAME) and/or atropine. In vivo motility was assessed by measurement of the colonic bead latency time. Randomly occurring ex vivo contractions appeared starting at P5. Starting at P14, rhythmic phasic contractions occurred whose frequency and amplitude increased over time. In vivo, bead latency was significantly reduced between P14 and P21. Ex vivo, EFS-induced contractile responses increased significantly over time and were significantly reduced by atropine starting at P14 but were sensitive to L-NAME only after P21. The proportion of ChATimmunoreactive (IR) neurons increased time dependently starting at P14. The proportion of nNOS-IR neurons increased as early as P5 compared with P1 but did not change afterward. Our data support a key role for cholinergic myenteric pathways in the development of postnatal motility and further identify them as putative therapeutic target for the treatment of GI motility disorders in the newborn. postnatal development; enteric nervous system; myenteric plexus; rat; motility; neonates THE POSTNATAL PERIOD is a key period of life and is particularly sensitive to the influence of various environmental factors. This period is characterized by the maturation of various organs and in particular of the gut. Indeed, various gastrointestinal (GI) functions such as intestinal barrier function or motility continue their maturation and development after birth. This is particularly true in rodents such as rats or mice, making them good models for studying GI dysfunctions observed in preterm infants, such as constipation (2).Among the key regulators of GI functions is the enteric nervous system (ENS). The ENS has been shown to control GI motility and intestinal barrier functions (22). Cholinergic excitatory motor neurons [identified as choline acetyltransferase (ChAT)-immunoreactive (IR)] and often colocalized with substance P and nitrergic inhibitory motor neurons [identified as neuronal nitric ox...