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  • Marker-based assays for studying gut transit in gnotobiotic and conventional mouse models.

Marker-based assays for studying gut transit in gnotobiotic and conventional mouse models.

STAR protocols (2021-11-23)
Sean T Koester, Naisi Li, Daniel M Lachance, Neelendu Dey
ABSTRACT

Gastrointestinal motility is regulated by a variety of environmental factors including gut microbes and metabolites. The ability to interrogate mouse models of gut motility has enabled elucidation of these relationships. Here we describe integration of the red carmine dye and fluorescence isothiocyanate-dextran marker-based assays for characterizing gut transit with spatial resolution, along with an optional intracolonic infusion protocol for studying the effects of metabolites on colonic transit. These protocols can be adapted for use in gnotobiotic and conventional mouse models. For complete details on the use and execution of this protocol, please refer to Li et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fluorescein isothiocyanate–dextran, average mol wt 70,000, (FITC:Glucose = 1:250)
Sigma-Aldrich
Cholic acid, from bovine and/or ovine, ≥98%
Sigma-Aldrich
Carmine, powder, ≥42.0% (UV-vis)
Sigma-Aldrich
Lithocholic acid, ≥95%
Sigma-Aldrich
Methyl cellulose, viscosity: 4,000 cP
Sigma-Aldrich
Taurocholic acid sodium salt hydrate, ≥95% (HPLC)
Sigma-Aldrich
β-Muricholic Acid, ≥98% (HPLC)
Sigma-Aldrich
Deoxycholic acid, ≥99.0% (T)