2022/05/24 by Cualcialpud Serna, Julian David, de Miranda Ramos, Vitor, Cabral-Costa, João Victor +5
paper · doi:10.26124/mitofit:2022-0021
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.