Molecular visualizing and quantifying immune-associated peroxynitrite fluxes in phagocytes and mouse inflammation model.

Abstract

Reactions of peroxynitrite (ONOO) with biomolecules can lead to cytotoxic and cytoprotective events. Due to the difficulty of directly and unambiguously measuring its levels, most of the beneficial effects associated with ONOOin vivo remain controversial or poorly characterized. Recently, optical imaging has served as a powerful noninvasive approach to studying ONOOin living systems. However, ratiometric probes for ONOOare currently lacking. Herein, we report the design, synthesis, and biological evaluation of F, a novel fluorescence indicator that relies on ONOO-induced diene oxidation. The remarkable sensitivity, selectivity, and photostability of Fenabled us to visualize basal ONOOin immune-stimulated phagocyte cells and quantify its generation in phagosomes by high-throughput flow cytometry analysis. With the aid of in vivo ONOOimaging in a mouse inflammation model assisted by F, we envision that Fwill find widespread applications in the study of the ONOObiology associated with physiological and pathological processes in vitro and in vivo.