Free Radical Research

Mitochondria are one of the major sources of reactive oxygen species (ROS) in mammalian cells. The generation of ROS underlies many physiological and pathophysiological processes that occur within cellular systems. Superoxide () is the proximal ROS generated during electron ‘leakage’ from the mitochondrial electron transport chain (mETC) and is known to be released at mitochondrial complex I and complex III. Monitoring mitochondrial production directly and in real-time offers the potential to improve understanding of the complex mechanisms involved during mitochondrial generation. This study reports the novel application of a cytochrome c functionalized amperometric sensor for monitoring generation in isolated mitochondrial fractions. The non-invasive sensor system described allowed a comparison of production following specific inhibition of complex I and complex III of the mETC to be made directly and in real-time.