Frontiers in Chemistry (Feb 2020)

Sensing Peroxynitrite in Different Organelles of Murine RAW264.7 Macrophages With Coumarin-Based Fluorescent Probes

  • Maria Weber,
  • Maria Weber,
  • Namiko Yamada,
  • Xue Tian,
  • Steven D. Bull,
  • Masafumi Minoshima,
  • Kazuya Kikuchi,
  • Kazuya Kikuchi,
  • Amanda B. Mackenzie,
  • Amanda B. Mackenzie,
  • Tony D. James

DOI
https://doi.org/10.3389/fchem.2020.00039
Journal volume & issue
Vol. 8

Abstract

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The elucidation of biological processes involving reactive oxygen species (ROS) facilitates a better understanding of the underlying progression of non-communicable diseases. Fluorescent probes are a powerful tool to study various ROS and have the potential to become essential diagnostic tools. We have developed a series of coumarin fluorescent probes for the selective and sensitive detection of peroxynitrite (ONOO−), a key ROS. Coumarin based probes exhibit good photostability, large Stokes shift and high quantum yields. The three ratiometric probes all contain a boronate ester motif for the detection of ONOO− and a distinctive organelle targeting group. The study of ONOO− generation in a particular organelle will allow more precise disease profiling. Hence, targeting groups for the mitochondria, lysosome and endoplasmic reticulum were introduced into a coumarin scaffold. The three ratiometric probes displayed sensitive and selective detection of ONOO− over other ROS species. All three coumarin probes were evaluated in murine RAW264.7 macrophages for detection of basal and stimulated ONOO− formation.

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