Dec 21, 2023
Whole Gut Transit Time, Fecal Water Content, and Fecal Output
- Jessica Griffiths1,
- Bryan Yoo1,
- Sarkis Mazmanian1
- 1California Institute of Technology
Protocol Citation: Jessica Griffiths, Bryan Yoo, Sarkis Mazmanian 2023. Whole Gut Transit Time, Fecal Water Content, and Fecal Output. protocols.io https://dx.doi.org/10.17504/protocols.io.36wgq3p1xlk5/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: 92590
Funders Acknowledgement:
Center for Enironmental and Microbial Interactions
Grant ID: n/a
Aligning Science Across Parkinson's
Grant ID: ASAP-020495
Heritage Medical Research Institute
Grant ID: n/a
Aligning Science Across Parkinson's
Grant ID: ASAP-000325
Department of Defense
Grant ID: PD160030
National Institutes of Health
Grant ID: GM007616 and DK078938
Emerald Foundation
Grant ID: n/a
Abstract
This protocol was used in "Peripheral Neuronal Activation Shapes the Microbiome and Alters Gut Physiology"
Experiment Preparation
Experiment Preparation
1d
6% (w/v) carmine red (Sigma-Aldrich, St. Louis, MO) with 0.5% methylcellulose (Sigma-Aldrich)
was dissolved and autoclaved prior to use.
1d
Experiment
Experiment
Mice were administered C21 (3 mg/kg) intraperitoneally, and subsequently orally
gavaged with 150 µL of carmine red solution.
5m
Following gavage, mice were single-housed with no bedding for the duration of the experiment, and
animals were not fasted beforehand.
Over the 5 hours following gavage, the time of expulsion was recorded for each fecal
pellet, and each pellet was collected in pre-weighed, 1.5 mL microcentrifuge
tube.
5h
Each pellet collected was checked for the presence of carmine red, and the time of
initial carmine red pellet expulsion was recorded as GI transit time.
Sample Processing
Sample Processing
2d
The mass of collected fecal pellets was determined, and pellets were left to dry in
an 80 ºC oven for 2 days before weighing the desiccated pellets and calculating
the pellets’ initial water content.
2d
Data Analysis
Data Analysis
1h
Fecal output rate for each mouse was calculated as the total number of pellets
expelled during the 5 hour time course post-C21 administration divided by the
time the last fecal pellet was expelled.
1h