Abstract
The enteric nervous system (ENS) orchestrates a broad range of important gastrointestinal functions such as intestinal motility and gastric secretion. The ENS can be affected by environmental factors, diet and disease. Changes due to these alterations are often hard to evaluate in detail when whole gut samples are used. Analyses based on pure ENS tissue can more effectively reflect the ongoing changes during pathological processes. Here, we present an optimized approach for the isolation of pure myenteric plexus (MP) from adult mouse and human. To do so, muscle tissue was individually digested with a purified collagenase. After incubation and a gentle mechanical disruption step, MP networks could be collected with anatomical integrity. These tissues could be stored and used either for immediate genomic, proteomic or in vitro approaches, and enteric neurospheres could be generated and differentiated. In a pilot experiment, the influence of bacterial lipopolysaccharide on human MP was analyzed using 2-dimensional gel electrophoresis. The method also allows investigation of factors that are secreted by myenteric tissue in vitro. The isolation of pure MP in large amounts allows new analytical approaches that can provide a new perspective in evaluating changes of the ENS in experimental models, human disease and aging.
Highlights
Correspondence and requests for materials should be addressed to Isolation of high-purity myenteric plexus from adult human and mouse gastrointestinal tract
Adult neurogenesis occurs in the mouse intestine in vivo in response to 5-Hydroxytriptamine 4 (5HT4) receptor activation or intestinal disruption and injury when glial cells transdifferentiate into neurons[20,21,22]
The tissues from the individual gut segments vary significantly, the muscle layer can be removed in all segments of the gastrointestinal tract (GIT)
Summary
Correspondence and requests for materials should be addressed to Isolation of high-purity myenteric plexus from adult human and mouse gastrointestinal tract. The isolation of pure MP in large amounts allows new analytical approaches that can provide a new perspective in evaluating changes of the ENS in experimental models, human disease and aging. The ENS is responsible for the regulation of homeostatic key features in the gastrointestinal tract, such as gastric secretion, absorption and motility[1,2] It consists of several intricate neural networks within the gut wall, which harbors up to several hundred million neurons and enteric glial cells, as well as neuronal progenitor cells, throughout its lifetime[3,4,5]. Enteric glial cells consist of several subpopulations that can be discriminated by their anatomical localization, morphologies or marker proteins such as GFAP, S100b, etc Their divergent functional properties have recently begun to be examined in detail[3]. As recently shown in the adult mouse model, enteric glia cells can proliferate during both steady state and after intestinal injury in vivo[20,21]
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