A Nuclear-Localized Fluorescent Hydrogen Peroxide Probe for Monitoring Sirtuin-Mediated Oxidative Stress Responses In Vivo

SummaryHydrogen peroxide (H2O2) can serve as a beneficial signaling agent or toxin depending on its concentration and location within a cell or organism. Methods to measure the localized accumulation of H2O2 in living specimens remain limited. Motivated to meet this need, we have developed a nuclear-localized fluorescent probe for H2O2, Nuclear Peroxy Emerald 1 (NucPE1), to selectively interrogate ROS fluxes within this sensitive organelle. NucPE1 selectively accumulates in the nuclei of a variety of mammalian cell lines as well as in whole model organisms like Caenorhabditis elegans, where it can respond to subcellular changes in H2O2 fluxes. Moreover, in vivo NucPE1 imaging reveals a reduction in nuclear H2O2 levels in worms overexpressing sir-2.1 compared with wild-type congeners, supporting a link between this longevity-promoting sirtuin protein and enhanced regulation of nuclear ROS pools.

Graphical AbstractFigure optionsDownload high-quality image (211 K)Download as PowerPoint slideHighlights
► NucPE1 selectively responds to H2O2 with a turn-on fluorescent response
► NucPE1 can detect H2O2 in the nuclei of a variety of common cell lines
► NucPE1 can detect H2O2 in vivo within the nuclei of C. elegans
► C. elegans overexpressing sir-2.1 have enhanced nuclear protection against H2O2