Abstract : Mitochondrial Ca2+ homeostasis is crucial for regulating vital functions such as respiration or apoptosis. Targeted aequorins are excellent probes to measure subcellular Ca2+. Ca2+ concentration in mitochondria ([Ca2+]M) is low at rest (about 10-7 M) and can increase to the micromolar or even approach the millimolar range, upon cell activation. Here we describe a new quantitative luminescent protocol to directly measure mitochondrial Ca2+ uptake, optimized for high throughput. The sensitivity of the method allows detection of changes in either the capacity or the affinity of mitochondrial Ca2+ transport.
Maria Teresa Alonso, Paloma Navas-Navarro, Javier Garcia-Sancho. A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake. Methods in Molecular Biology, 1567, pp.245-253, 2017, ⟨10.1007/978-1-4939-6824-4_15⟩. ⟨hal-02179310⟩