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

Publication date: 15 April 2017
Source:Biosensors and Bioelectronics, Volume 90
Author(s): Zan Li, Shi-Hai Yan, Chen Chen, Zhi-Rong Geng, Jia-Yin Chang, Chun-Xia Chen, Bing-Huan Huang, Zhi-Lin. Wang
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 ONOO⁻ in vivo remain controversial or poorly characterized. Recently, optical imaging has served as a powerful noninvasive approach to studying ONOO⁻ in living systems. However, ratiometric probes for ONOO⁻ are currently lacking. Herein, we report the design, synthesis, and biological evaluation of F482, a novel fluorescence indicator that relies on ONOO⁻-induced diene oxidation. The remarkable sensitivity, selectivity, and photostability of F482 enabled us to visualize basal ONOO⁻ in immune-stimulated phagocyte cells and quantify its generation in phagosomes by high-throughput flow cytometry analysis. With the aid of in vivo ONOO⁻ imaging in a mouse inflammation model assisted by F482, we envision that F482 will find widespread applications in the study of the ONOO⁻ biology associated with physiological and pathological processes in vitro and in vivo.



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