Hypoxic-Ischemic Injury in the Developing Brain: The Role of Reactive Oxygen Species Originating in Mitochondria

Figure 2

(a) H₂O₂ emission rate from brain mitochondria isolated from p10 naïve mouse and supported either with succinate (FAD-linked substrate) or malate-glutamate (NAD-linked substrates). Time-points when mitochondria (mito, 0.05 mg/mL) or rotenone (1 μM) were added are indicated. Cerebral nonsynaptic mitochondria were isolated and mitochondrial H₂O₂ fluorescence was measured using Amplex-ultra-red and horse radish peroxidase assay as described in [31]. (b–e), a schematic mechanism for ROS generation in MRC fueled with NAD-linked substrate, before (b) and after inhibition of complex I with rotenone (c), or FAD-linked substrate, before (d) and after rotenone supplementation (e). RET: reverse electron trasport, FET: forward electron transport.