Serna 2022 MitoFit

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Serna 2022 MitoFit

Publications in the MiPMap
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.

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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:

Graphical abstract

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


ORCID:ORCID.png Serna Julian DC, ORCID.png Ramos Vitor M, ORCID.png Cabral-Costa Joao V, ORCID.png Vilas-Boas Eloisa A, ORCID.png Amaral Andressa G, ORCID.png Ohya Georgia, ORCID.png da Silva Camille CC, ORCID.png Kowaltowski Alicia J

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 

Bioblast 2022