In vitro and in vivo imaging of peroxynitrite by a ratiometric boronate-based fluorescent probe

Biosens Bioelectron. 2017 May 15:91:849-856. doi: 10.1016/j.bios.2017.01.027. Epub 2017 Jan 23.

Abstract

Peroxynitrite (ONOO-) is an important species involved in many physiopathological processes. Progresses have been made in developing novel fluorescent probes to detect peroxynitrite with relatively high sensitivity and specificity. Herein, we report the synthesis, characterization and biological applications of a new boronate-based fluorescent probe, 4-MB. The studies showed that 4-MB exhibits a dual ratiometric and calorimetric response toward peroxynitrite due to ONOO--triggered oxidative reaction. A possible mechanism of the oxidation reaction was proposed and the reaction product was isolated and characterized using different spectroscopic methods. We have thoroughly demonstrated the utility of 4-MB for intracellular peroxynitrite imaging. Further, we showed that 4-MB can be potentially employed to visualize exogenous and endogenous peroxynitrite in RAW264.7 macrophages, EAhy926 cells, zebrafish and in live tissues from a high-fat diet-induced obese mouse model.

Keywords: Fluorescence; Imaging; Peroxynitrite; Sensor.

MeSH terms

  • Animals
  • Biosensing Techniques / methods
  • Boronic Acids / chemistry*
  • Cell Line
  • Fluorescent Dyes / chemistry*
  • Macrophages / chemistry
  • Macrophages / cytology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Obesity / pathology
  • Optical Imaging / methods*
  • Peroxynitrous Acid / analysis*
  • RAW 264.7 Cells
  • Spectrometry, Fluorescence / methods

Substances

  • Boronic Acids
  • Fluorescent Dyes
  • Peroxynitrous Acid