Krycer 2017 Biochem Biophys Res Commun
|Krycer JR, Fisher-Wellman KH, Fazakerley DJ, Muoio DM, James DE (2017) Bicarbonate alters cellular responses in respiration assays. Biochem Biophys Res Commun 489:399-403.|
Abstract: Metabolic assay buffers often omit bicarbonate, which is susceptible to alkalinisation in an open environment. Here, we assessed the effect of including bicarbonate in respirometry experiments. By supplementing HEPES-buffered media with low concentrations of bicarbonate, we found increased respiration in adipocytes and hepatocytes, but not myotubes. This was observed across multiple respirometry platforms and was independent of effects on enhanced insulin sensitivity, pH drift, or mitochondrial function. Permeabilised cell experiments suggest that bicarbonate increases substrate availability, likely by acting as a cofactor for carboxylase enzymes. This emphasises the importance of buffer choice in experimental biology.
Copyright © 2017 Elsevier Inc. All rights reserved. • Keywords: Bicarbonate, Buffer, HEPES, Mitochondria, Respiration • Bioblast editor: Plangger M • O2k-Network Lab: AU Sydney James D, US NC Durham Koves TR
Labels: MiParea: Respiration
Organism: Mouse Tissue;cell: Liver, Fat Preparation: Permeabilized cells, Intact cells
Coupling state: LEAK, OXPHOS Pathway: N, NS HRR: Oxygraph-2k