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.
Since several years, the enteric nervous system (ENS) is getting more and more in the focus of gastrointestinal research. While the main interest was credited for years to the enteric neurons and their functional properties, less attention has been paid on the enteric glial cells (EGCs). Although the similarity of EGCs to central nervous system (CNS) astrocytes has been demonstrated a long time ago, EGCs were investigated in more detail only recently. Similar to the CNS, there is not "the" EGC, but also a broad range of diversity. Based on morphology and protein expression, such as glial fibrillary acidic protein (GFAP), S100, or Proteolipid-protein-1 (PLP1), several distinct glial types can be differentiated. Their heterogeneity in morphology, localization, and transcription as well as interaction with surrounding cells indicate versatile functional properties of these cells for gut function in health and disease. Although NG2 is found in a subset of CNS glial cells, it did not colocalize with the glial marker S100 or GFAP in the ENS. Instead, it in part colocalize with PDGFRα, as it does in the CNS, which do stain fibroblast-like cells in the gastrointestinal tract. Moreover, there seem to be species dependent differences. While GFAP is always found in the rodent ENS, this is completely different for the human gut.Only the compromised human ENS shows a significant amount of GFAPpositive glial cells. So, in general we can conclude that the EGC population is species specific and as complex as CNS glia. Anat Rec, 302:1333Rec, 302: -1344Rec, 302: , 2019.
The enteric nervous system has to adapt to altering dietary or environmental conditions and presents an enormous plasticity that is conserved over the whole lifespan. It harbours neural-crest-derived neurons, glial cells and their precursors. Based on a nestin-green fluorescent protein (NGFP) transgenic model, a histological inventory has been performed to deliver an overview of neuronal and glial markers for the various parts of the gastrointestinal tract in newborn (postnatal day 7) and adult mice under homeostatic conditions. Whereas NGFP-positive glial cells can be found in all parts of the gut at any individual age, a specific NGFP population is present with both neuronal morphology and marker expression in the myenteric plexus (nNGFP). These cells appear in variable quantities, depending on age and location. Their overall abundance decreases from newborn to adults and their spatial distribution is also age-dependent. In newborn gut, nNGFP cells are found in similar quantities throughout the gut, with a significantly lower presence in the duodenum. Their expression increases in the adult mouse from the stomach to the colon. All of these nNGFP cells expressed either (but not both) of the glia markers S100 or glial fibrillary acidic protein (GFAP). In the S100-positive glia population, a subset of cells also shows a neuronal morphology (nS100), without expressing nestin. Thus, the presence of premature neurons that express NGFP demonstrates that neurogenesis takes place far beyond birth. In enteric neurons, NGFP acts as a marker for neuronal plasticity showing the differentiation and change in the phenotype of neuronal precursor cells.
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