Review: Using isolated mitochondria to investigate mitochondrial hydrogen peroxide metabolism

Mitochondria are recognized as centrally important to cellular reactive oxygen species (ROS), both as a potential source and due to their substantial antioxidant capacity. While much of the initial ROS formed by mitochondria is superoxide, this is rapidly converted to hydrogen peroxide (H₂O₂) which more readily crosses membranes making H₂O₂ important in both redox signalling mechanisms and conditions of oxidative stress. Here I outline our studies on mitochondrial H₂O₂ metabolism with a focus on some of the challenges and strategies involved with developing an integrated model of mitochondria being intrinsic regulators of H₂O₂. This view of mitochondria as regulators of H₂O₂ goes beyond the simpler contention of them being net producers or consumers. Moreover, the integration of both consumption and production can then be tied to a putative mechanism linking energy sensing at the level of the mitochondrial protonmotive force. This mechanism would provide a means of mitochondria communicating their energetic status the extramitochondrial compartment via local H₂O₂ concentrations. I conclude by explaining how these concepts developed using rodent muscle as a model have high relevance and applicability to comparative studies.