Figure 5

The simultaneous measurement of cytosolic (Fura-2) and mitochondrial (Rhod-2) calcium concentrations in WT and G93A transfected transfected SH-SY5Y cells during FCCP-evoked mitochondrial Ca²⁺ release. A) The kinetic profile of the FCCP-evoked Ca²⁺ release in the WT transfected SH-SY5Y neuroblastoma cells; the cytosolic (Error bar green, black square trace) and mitochondrial (Error bar red, black circle trace) compartment were measured simultaneously. The trace represents the mean of 5 cells in focus stimulated with 2µM FCCP (5 point smoothing). B) The corresponding kinetic profile of the FCCP-evoked Ca²⁺ release in the G93A transfected SH-SY5Y neuroblastoma cells; the cytosolic (Error bar green, black square trace) and mitochondrial (Error bar red, black circle trace) compartment were measured simultaneously. The trace represents the mean of 5 cells in focus stimulated with 2µM FCCP (5 point smoothing). FCCP-evoked [Ca²⁺]mito signals were smaller in amplitude and exhibited slower kinetics in G93A transfected SH-SY5Y cells compared to WT transfected cells and were altered from [Ca²⁺]i efflux. C) A bar diagram of the cytosolic (green bar) and mitochondrial (red bar) fluorescence signals (F/F0) from WT (F/F0 = 0.1569 ± 0.0235 for [Ca²⁺]i and F/F0 = -0.1069 ± 0.0181 for [Ca²⁺]mito; hollow; N=5, n=17) and G93A (F/F0 = 0.1008 ± 0.0248 for [Ca²⁺]i and F/F0 = -0.0486 ± 0.0043 for[Ca²⁺]mito; striped pattern, N=4; n=17) transfected SH-SY5Y neuroblastoma cells. Values represent means ± SD, **p<0.001. N= Number of experiments; n= Number of cells.