Serna 2022 MitoFit
| 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. MitoFit Preprints 2022.21. https://doi.org/10.26124/mitofit:2022-0021 |
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Measuring mitochondrial Ca2+ efflux in isolated mitochondria and permeabilized cells
Serna Julian DC, Ramos Vitor M, Cabral-Costa Joao V, Vilas-Boas Eloisa A, Amaral Andressa G, Ohya Georgia, da Silva Camille CC, Kowaltowski Alicia J (2022-05-24) MitoFit Prep
Abstract:
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
ORCID:
Serna Julian DC, Ramos Vitor M, Cabral-Costa Joao V, Vilas-Boas Eloisa A, Amaral Andressa G, Ohya Georgia, da Silva Camille CC, Kowaltowski Alicia J
Labels: MiParea: Exercise physiology;nutrition;life style, Pharmacology;toxicology
Bioblast 2022
