RNA-Seq of FACS sorted Enteric Glia cells from the LMMP after T-cell activation via CD3

The Experiment was performed to understand the changes in expression profile of enteric glia cells in context of intestinal inflammation. Enteric glia are crucial regulators of gastrointestinal motility, however enteric glia alterations in IBD are poorly characterized. Acute T-cell activation rapidly impacts enteric glia activation, and induces cell death pathways, which negatively impacts small intestinal transit. This experiment has been crucial to identify IFN?- and TNFa -driven necrosis in activated glia cell subsets. In order to sequence pure enteric glia, a mouse strain of Bl6 mice with a Plp1CreERT insert for glial specificity crossed with mice with a flox controlled tdTomato fluorescent protein on the ROSA26 Locus. The mice received a peritoneal injection of an anti-CD3 antibody or its isotype control antibody. At 6 hours, the small intestine was collected and the longitudinal muscle layer with the myenteric plexus stripped off. After enzymatic dissociation and trituration, the enteric glia were FACS sorted and finally the RNA isolated.