Goudie 2022 Sci Rep

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Goudie L, Mancini NL, Shutt TE, Holloway GP, Mu C, Wang A, McKay DM, Shearer J (2022) Impact of experimental colitis on mitochondrial bioenergetics in intestinal epithelial cells. https://doi.org/10.1038/s41598-022-11123-w

Β» Sci Rep 12:7453. PMID: 35523978 Open Access

Goudie Luke, Mancini Nicole L, Shutt Timothy E, Holloway Graham P, Mu Chunglong, Wang Arthur, McKay Derek M, Shearer Jane (2022) Sci Rep

Abstract: Intestinal homeostasis is highly dependent on optimal epithelial barrier function and permeability. Intestinal epithelial cells (IEC) regulate these properties acting as cellular gatekeepers by selectively absorbing nutrients and controlling the passage of luminal bacteria. These functions are energy demanding processes that are presumably met through mitochondrial-based processes. Routine methods for examining IEC mitochondrial function remain sparse, hence, our objective is to present standardized methods for quantifying mitochondrial energetics in an immortalized IEC line. Employing the murine IEC4.1 cell line, we present adapted methods and protocols to examine mitochondrial function using two well-known platforms: the Seahorse Extracellular Flux Analyzer and Oxygraph-2 k. To demonstrate the applicability of these protocols and instruments, IEC were treated with and without the murine colitogenic agent, dextran sulfate sodium (DSS, 2% w/v). Profound impairments with DSS treatment were found with both platforms, however, the Oxygraph-2 k allowed greater resolution of affected pathways including short-chain fatty acid metabolism. Mitochondrial functional analysis is a novel tool to explore the relationship between IEC energetics and functional consequences within the contexts of health and disease. The outlined methods offer an introductory starting point for such assessment and provide the investigator with insights into platform-specific capabilities.

β€’ Bioblast editor: Plangger M β€’ O2k-Network Lab: Holloway Graham P, CA Calgary Shearer J

Labels: MiParea: Respiration, Instruments;methods 

Organism: Mouse  Tissue;cell: Endothelial;epithelial;mesothelial cell  Preparation: Permeabilized cells 

Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, CIV, ROX  HRR: Oxygraph-2k 

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