Synergistic Triggering of Superoxide Flashes by Mitochondrial Ca2+ Uniport and Basal Reactive Oxygen Species Elevation

Mitochondrial superoxide flashes reflect a quantal, bursting mode of reactive oxygen species (ROS) production that arises from stochastic, transient opening of the mitochondrial permeability transition pore (mPTP) in many types of cells and in living animals. However, the regulatory mechanisms and the exact nature of the flash-coupled mPTP remain poorly understood. Here we demonstrate a profound synergistic effect between mitochondrial Ca 2+ uniport and elevated basal ROS production in triggering superoxide flashes in intact cells. Hyperosmotic stress potently augmented the flash activity while simultaneously elevating mitochondrial Ca 2+ and ROS. Blocking mitochondrial Ca 2+ transport by knockdown of MICU1 or MCU, newly identified components of the mitochondrial Ca 2+ uniporter, or scavenging mitochondrial basal ROS markedly diminished the flash response. More importantly, whereas elevating Ca 2+ or ROS production alone was inefficacious in triggering the flashes, concurrent physiological Ca 2+ and ROS elevation served as the most powerful flash activator, increasing the flash incidence by an order of magnitude. Functionally, superoxide flashes in response to hyperosmotic stress participated in the activation of JNK and p38. Thus, physiological levels of mitochondrial Ca 2+ and ROS synergistically regulate stochastic mPTP opening and quantal ROS production in intact cells, marking the flash as a coincidence detector of mitochondrial Ca 2+ and ROS signals. Background: Superoxide flashes are newly discovered elemental mitochondrial ROS signaling events and reflect transient openings of mPTP. Results: Mitochondrial Ca 2+ uniport and basal ROS elevation synergistically trigger superoxide flashes. Conclusion: Superoxide flashes are regulated by physiological levels of Ca 2+ and ROS. Significance: Demonstrating Ca 2+ - and ROS-induced flash activation sheds new light on physiological regulation and possible roles of mPTP.