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Fig. 6. Mitochondrial flux-force-structure diagram. (A-C) The energy state was modulated in living cells grown in a galactose medium (a), by three different means, which include: OXPHOS uncoupling with CCCP (b,c), respiratory chain inhibition with rotenone (d-f) and inhibition of mitochondrial ATP synthesis with oligomycin (g,h). All chemicals were added to the culture medium and incubated at 37°C. Rotenone was used for 4 hours at 2.4, 3.4 (d), 6.9 (e), 9.6 and 12 ng/ml (f). CCCP was used for 30 minutes at 5 and 10 µM (b), and 15 and 20 µM (c). Oligomycin was used at 0.1 and 0.2 ng/ml (g), and 0.5 ng/ml (h) for 3 hours. Staurosporin was used at 1 µM for 6 hours (k). Results for cells grown in glucose medium are also shown (a'). (A) Diagram showing the variation of mitochondrial respiration as a function of 
, expressed as a percentage of the control value and measured on MRC5 cells grown in galactose. (B) ROS steady-state level measured in the cytosol (black bars) or the mitochondrion (gray bars). Letters refer to the experimental conditions described in A. (C) mt-network organization observed in the above described situations labeled a-k. Also shown is the mt-network organization of cells taken from patients with (i) a severe complex-I defect and (j) multiple respiratory-chain deficiency. These cells lines are also positioned on the diagram shown in A, according to their bioenergetic coordinates. Fragmentation of the mt-network was also observed in MRC5 cells treated for 6 hours with staurosporin 1 µM (k). In addition, we discovered the details of mitochondrial ultrastructure in cells treated with rotenone (1) and CCCP (2), compared with the control (3).
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