Quantitative gene expression in gastrointestinal tissues following consistent activation of ChAT or TH enteric neurons versus non-activated control mice

Tools to study, and knowledge of, enteric nervous system development and function lag behind brain research. Herein, we deploy recombinant adeno-associated viral (rAAV) vectors with enhanced tropism for the gut to map and activate enteric neurons in mice with spatial and temporal resolution. we employed chemogentics to specifically activate gut neurons that express choline acetyltransferase (ChAT+) or tyrosine hydroxylase (TH+). Targeted activation of ChAT+ or TH+ neuronal populations associated with the gastrointestinal (GI) tract altered the intestinal transcriptome. ChAT-Cre or TH-Cre mice (6-8 weeks of age) were infected with activating DREADDs or a control peripheral nervous system virus expressing a reporter protein instead (AAV-PHP.S:hSYN1-DIO-mRuby2). C21 was administered intraperitoneally (i.p.), once daily for 10 consecutive days to all mice. One hour following the last C21 injection (when mice are 10-13 weeks of age), 1cm of tissue from the distal SI and proximal colon was harvested, and gene expression profiled by QuantSeq, a quantitative 3’ mRNA-sequencing technology.