A tandem activity-based sensing and labeling strategy enables imaging of transcellular hydrogen peroxide signaling.
Reactive oxygen species (ROS) like hydrogen peroxide (H2O2) are transient species that have broad actions in signaling and stress, but spatioanatomical understanding of their biology remains insufficient. Here, we report a tandem activity-based sensing and labeling strategy for H2O2 imaging that enables capture and permanent recording of localized H2O2 fluxes. Peroxy Green-1 Fluoromethyl (PG1-FM) is a diffusible small-molecule probe that senses H2O2 by a boronate oxidation reaction to trigger dual release and covalent labeling of a fluorescent product, thus preserving spatial information on local H2O2 changes. This unique reagent enables visualization of transcellular redox signaling in a microglia-neuron coculture cell model, where selective activation of microglia for ROS production increases H2O2 in nearby neurons. In addition to identifying ROS-mediated cell-to-cell communication, this work provides a starting point for the design of chemical probes that can achieve high spatial fidelity by combining activity-based sensing and labeling strategies.
['Iwashita|Hidefumi|H|0000-0001-6709-0897', 'Castillo|Erika|E|0000-0003-2492-901X', 'Messina|Marco S|MS|0000-0003-2827-118X', 'Swanson|Raymond A|RA|0000-0002-3664-5359', 'Chang|Christopher J|CJ|0000-0001-5732-9497']
D000818:Animals D001897:Boronic Acids D002450:Cell Communication D002540:Cerebral Cortex D018920:Coculture Techniques D004622:Embryo, Mammalian D005456:Fluorescent Dyes D006367:HeLa Cells D006801:Humans D006861:Hydrogen Peroxide D051379:Mice D017628:Microglia D015335:Molecular Probes D009474:Neurons D010084:Oxidation-Reduction D010269:Paraquat D000067996:RAW 264.7 Cells D015398:Signal Transduction D013194:Staining and Labeling
activity-based sensing redox signaling NADPH oxidase fluorescent hydrogen peroxide probe oxidative stress