Neural and glial phenotypic expression by neural crest cells in culture: Effects of control and presumptive aganglionic bowel from ls/ls mice

Abstract

The enteric nervous system is formed by cells that migrate to the bowel from the neural crest. Previous experiments have established that avian crest cells in vitro will colonize explants of murine bowel and there give rise to neurons. It has been proposed that phenotypic expression by the crest‐ derived precursors of enteric neurons and glia is critically influenced by the microenvironment these cells encounter within the gut. To test this hypothesis, quail crest cells were cocultured with explants of control or presumptive aganglionic bowel from the ls/ls mutant mouse, and the effects of the enteric tissue on five phenotypic markers of crest cell development were followed. Aganglionosis develops in the terminal region of the colon of the ls/ls mouse because viable crest‐ derived neural and glial precursors fail to colonize this tissue. Expression of the phenotypic markers in the cocultures was compared with that in cultures of crest alone, crest plus neural tube, and gut grown alone. The markers examined were melanogenesis and immunostaining with antisera to 5‐ hydroxytryptamine (5‐ HT) and tyrosine hydroxylase (TH) and the monoclonal antibodies, NC‐ 1 and GlN1. Explants of control, but not presumptive aganglionic ls/ls gut were found to increase the incidence of the expression of 5‐ HT and NC‐ 1 immunoreactivities; moreover, especially near the gut, the assumption of a neuronal morphology by 5‐ HT‐, NC‐1‐, and GlN1‐ immunoreactive cells was also increased. Coincidence of expression of 5‐ HT with NC‐ 1 and GlN1 immunoreactivities was observed. The effect of the bowel was selective in that the expression of TH immunoreactivity, which is not a marker of mature enteric neurons, was reduced rather than enhanced. The effect of enteric explants on crest cell development was specific in that it was not mimicked by explants of metanephros, which inhibited expression of 5‐ HT immunoreactivity and the acquisition of a neuritic form by NC‐ 1‐ immunoreactive cells. It is concluded that theenteric microenvironment affects the phenotypic expression of subsets of crest cells and that this action of the bowel. Is manifested in vitro. The inability of presumptive aganglionic gut from ls/ls mice to influence neural phenotypic expression may be due to the failure of this tissue to produce putative factor(s) required for theeffector to the inability of the crest‐ derived precursor cells to migrate into the abnormal enteric tissue.