Subcellular ROS imaging methods: Relevance for the study of calcium signaling

Abstract

Recent advances in genetically encoded fluorescent probes have dramatically increased the toolkit available for imaging the intracellular environment. Perhaps the biggest improvements have been made in sensing specific reactive oxygen species (ROS) and redox changes under physiological conditions. The new generation of probes may be targeted to a wide range of subcellular environments. By targeting such probes to compartments and organelle surfaces they may be exposed to environments, which support local signal transduction and regulation. The close apposition of the endoplasmic reticulum (ER) with mitochondria and other organelles forms such a local environment where Ca(2+) dynamics are greatly enhanced compared to the bulk cytosol. We describe here how newly developed genetically encoded redox indicators (GERIs) might be used to monitor ROS and probe their interaction with Ca(2+) at both global and local level.

Keywords: Grx1-roGFP2; H(2)O(2); HyPer; Redox; SypHer.