Rate of ROS production in mitochondria from control mice and after t-BH exposure.

<p>A-B: Representative raw traces of the ROS-recordings. The rate of hydrogen peroxide (H₂O₂) production was followed spectrofluorometrically by its reaction with Amplex Red in the presence of horseradish peroxidase. The rate of ROS production was recorded in three consecutive steps: After addition of mitochondria alone, after addition of substrates (5 mM glutamate and 5 mM malate) and after addition of one submaximal dose of Ca²⁺. Traces are shown under control conditions (no additions; black), in the presence of 500 µmol/L ADP (blue), 0.2 µmol/L CsA (green), and 1mmol/L ATP + 10 mmol/L creatine (Cr) (red). A: control mice; B: after exposure to 5 μmol/L t-BH. C-D: Column diagrams of the averaged results under the conditions indicated below each column. Steady-state H₂O₂ production was recorded in the mitochondria alone (rate between 1-3 min; white columns), after addition of substrates (rate between 6.5-8.5 min; light grey columns), and after addition Ca²⁺ (rate between 11-15 min; dark grey columns). Treatments within each group are indicated below the columns. All values are mean ± SEM. † and †† denote significant effect of substrates and/or Ca²⁺, P < 0.05, P < 0.01 respectively. *, ** denote significant difference P < 0.05, and P < 0.01, respectively, between control and different treatments within the group. <b><i>#</i></b>and ^## denote significant difference (P < 0.05 and P<0.01, respectively) between control and t-BH with the same treatment. The number of experiments is indicated below.