|Serna JDC, Ramos VM, Cabral-Costa JV, Vilas-Boas EA, Amaral AG, Ohya G, da Silva CCC, Kowaltowski AJ (2022) Measuring mitochondrial Ca2+ efflux in isolated mitochondria and permeabilized cells. https://doi.org/10.26124/mitofit:2022-0021|
» MitoFit Preprints 2022.21.
Serna 2022 Abstract Bioblast: Mitochondrial Ca2+ efflux is essential for mitochondrial and cell Ca2+ homeostasis. Mitochondrial inner membrane Ca2+/H+ and Na+/Li+/Ca2+ (NCLX) exchangers are known today to be plastic transporters, with important roles in physiological responses and pathological states. Until now, however, no consensus protocols were available to measure mitochondrial Ca2+ efflux, and we find that some published protocols may induce mitochondrial permeability transition, underestimating the effects of these exchangers. In this work we describe a method to measure Na+-sensitive and insensitive mitochondrial Ca2+ efflux activity in isolated mitochondria and permeabilized cells using the Ca2+ Green indicator and a fluorimeter. A checklist is provided to avoid artefacts as well as pinpoint adaptations necessary in specific experimental models. • Keywords: mitochondria, Ca2+ efflux, NCLX, mPTP, liver • Bioblast editor: Cecatto C • O2k-Network Lab: BR Sao Paulo Kowaltowski AJ
Labels: MiParea: Instruments;methods
Stress:Permeability transition Organism: Human, Mouse Tissue;cell: Liver, Other cell lines Preparation: Permeabilized cells, Isolated mitochondria
Regulation: Calcium, Ion;substrate transport Coupling state: LEAK, OXPHOS Pathway: S, ROX HRR: Oxygraph-2k