Environmental Effects on Reactive Oxygen Species Detection-Learning from the Phagosome.

Significance: Reactive oxygen species (ROS) fulfill numerous roles in biology ranging from signal transduction to the induction of cell death. To advance our understanding of these sometimes contradictory roles, quantitative, specific, and sensitive ROS measurements are required.

Recent Advances: Several organic or genetically encoded probes were successfully developed for ROS detection.

Critical Issues: In some cases, ROS production occurs in a harsh environment such as low pH or high concentration of proteases. However, the ROS sensor may be sensitive to such environmental conditions and therefore becomes inaccurate. While the sensitivity of many ROS sensors to pH is known, many other environmental conditions remain unexplored. This article illustrates the interference between ROS sensors and their environment using the phagosome as an example. In the phagosome, pH changes, high concentration of ROS, and the presence of many proteases generate a hostile and rapidly changing environment.

Future Directions: Difficulties due to cell movement and continuous formation of new phagosomes can be reduced by ratio measurements, if appropriate dyes are identified. For detection in live cells and subcellular locations, fluorescent proteins (FPs) offer several advantages and are used to create biosensors for ROS. Some FPs are directly sensitive to certain ROS as shown here. Although this may compromise their use in an environment with high levels of ROS, it can also be exploited for ROS measurement directly with the FPs themselves. For all types of ROS detection, we suggest a set of basic guidelines for testing the environmental sensitivity of an ROS sensor. Antioxid. Redox Signal. 25, 564-576.