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

Biosens Bioelectron. 2017 Apr 15:90:75-82. doi: 10.1016/j.bios.2016.11.036. Epub 2016 Nov 15.

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 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.

Keywords: Inflammation; Knoevenagel condensation; ONOO(−); Phagocytes; Quantifying; Visualizing.

MeSH terms

  • Animals
  • Biosensing Techniques*
  • Disease Models, Animal
  • Inflammation / diagnosis*
  • Inflammation / immunology
  • Mice
  • Peroxynitrous Acid / chemistry
  • Peroxynitrous Acid / isolation & purification*
  • Phagocytes / chemistry
  • Phagocytes / immunology

Substances

  • Peroxynitrous Acid