After sacrifice, the caudal half of the animal is pinned out. A ventral midline incision is made and the colon is cut at the cecum and separated from the small intestine, which is removed. Then the pelvic nerve is dissected by slowly removing all of the muscle bone and cartilage surrounding the distal colon. Leave the tuft of tiisue surrounding the pelvic nerve intact. For a good reference for the dissection, see: Feng and Gebhart, 2015, J Vis Exp,(95), 52310. The colon plus pelvic nerves are then placed into a sylguard-lined dissection dish filled with oxygenated Krebs-Ringers solution. The colon is then gently flushed of its contents. For all experiments, the colon is perfused with 35-degree, oxygenated Krebs-Ringers for the duration of the experiment. A suction electrode connected to a 10ml syringe and tubing is then placed over tuft of tissue with pelvic nerve, and upward pressure applied to suck in nerve.