Colonic Migrating Motor Complexes
Bryan Yoo, Jessica Griffiths, Sarkis Mazmanian
Abstract
Protocol used in "Peripheral Neuronal Activation Shapes the Microbiome and Alters Gut Physiology"
Steps
Sample Preparation
Intact colons were dissected from cervically-dislocated mice.
Colons were flushed and placed in pre-oxygenated (95% O2, 5% CO2)
Krebs-Henseleit solution at RT
Proximal and distal ends were cannulated to 2 mm
diameter tubes and secured in the center of an organ bath with continuously
oxygenated Krebs-Henseleit solution at 37 ºC.
Syringe pumps were connected to the
inlet and outlet tubes to maintain a flow of solution at a rate of 500 µL/min
through the colon.
The system was allowed to equilibrate for 30 minutes before
recording.
Data Aquisition
Baseline recordings were taken for 30 minutes, then the Krebs
solution in the organ bath was briefly removed, mixed with C21 to a final
concentration of 2 µM, and replaced in the organ bath. Recordings were taken
for another 30 minutes.
