Laser-capture microdissection for layer-specific analysis of enteric ganglia

Abstract

The enteric nervous system (ENS) is the division of the autonomic nervous system that innervates the gastrointestinal (GI) tract and controls central intestinal functions such as peristalsis and fluid movement. Enteric nerve cell bodies (neurons and glia) are predominantly organized in ganglionated networks that are present along the entire length of the GI tract in multiple tissue layers. Most cell bodies are organized in the myenteric plexus allocated between the longitudinal and the circular muscle layers or in the submucosal plexus between muscle tissue and mucosa. The site-specific characteristics of these enteric nerve cells have traditionally been analyzed via imaging techniques. Laser-capture microdissection (LCM) offers the prospect of site-specifically analyzing the gene expression profiles of these different subpopulations. This protocol addresses critical aspects of handling intestinal tissue for ENS dissection, such as the optimal quick-staining procedure, suitable laser settings, and limits of tissue material required to successfully dissect and analyze tissue layers for gene expression.