Oxidative Medicine and Cellular Longevity / 2015 / Article / Fig 3

Research Article

Increased Susceptibility of Gracilinanus microtarsus Liver Mitochondria to Ca2+-Induced Permeability Transition Is Associated with a More Oxidized State of NAD(P)

Figure 3

Effect of mitochondrial permeability transition (MPT) inhibitors on the Ca2+ retention capacity of mouse and marsupial liver mitochondria. Isolated mouse and marsupial liver mitochondria (LM; 0.5 mg/mL) were incubated in standard reaction medium supplemented with NADH-linked (2 mM malate, 1 mM pyruvate, 1 mM α-ketoglutarate, and 1 mM glutamate) respiratory substrates and 0.2 μM Calcium Green-5N. Some experiments were conducted in the presence of the MPT inhibitors: 1 μM cyclosporin A (CsA), 200 μM ADP, or 200 μM ATP plus 3 mM MgCl2, as indicated in the figure. (a) Representative experiments for estimation of calcium retention capacity of mouse and marsupial liver mitochondria in the presence of ADP. (b) To assess the mitochondrial Ca2+ retention capacity, pulses of Ca2+ (5 μM for control conditions or 30 μM for the conditions in the presence of MPT inhibitors CsA, ADP, or ATP plus Mg2+) were added until mitochondrial Ca2+ release occurred. The sum of Ca2+ pulses prior to MPT pore opening was taken as the mitochondrial Ca2+ retention capacity ( for all conditions except for “ATP+Mg2+,” where ). versus respective condition in mouse mitochondria.

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