Dec 21, 2023

Public workspaceColonic Migrating Motor Complexes

DOI
dx.doi.org/10.17504/protocols.io.n92ldm61nl5b/v1
  • 1California Institute of Technology
Jessica Griffiths
Open access
DOI: dx.doi.org/10.17504/protocols.io.n92ldm61nl5b/v1
Protocol Citation: Jessica Griffiths, Bryan Yoo, Sarkis Mazmanian 2023. Colonic Migrating Motor Complexes. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldm61nl5b/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 20, 2023
Last Modified: December 21, 2023
Protocol Integer ID: 92583
Funders Acknowledgement:
Center for Environmental and Microbial Interactions
Grant ID: n/a
Emerald Foundation
Grant ID: n/a
Heritage Medical Research Institute
Grant ID: n/a
Department of Defense
Grant ID: PD160030
National Institutes of Health
Grant ID: GM007616 and DF078938
Aligning Science Across Parkinson's
Grant ID: ASAP-000375
Aligning Science Against Parkinson's
Grant ID: ASAP-020495
Abstract
Protocol used in "Peripheral Neuronal Activation Shapes the Microbiome and Alters Gut Physiology"
Sample Preparation
Sample Preparation
1h 5m
Intact colons were dissected from cervically-dislocated mice.

10m
Colons were flushed and placed in pre-oxygenated (95% O2, 5% CO2) Krebs-Henseleit solution at RT
10m
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.
10m
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.
5m
The system was allowed to equilibrate for 30 minutes before recording.
30m
Data Aquisition
Data Aquisition
1h
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.
1h