Spatially-resolved Intracellular Sensing of Hydrogen Peroxide in Living Cells

Sci Rep. 2015 Nov 20;5:16929. doi: 10.1038/srep16929.

Abstract

Understanding intracellular redox chemistry requires new tools for the site-specific visualization of intracellular oxidation. We have developed a spatially-resolved intracellular sensor of hydrogen peroxide, HyPer-Tau, for time-resolved imaging in live cells. This sensor consists of a hydrogen peroxide-sensing protein tethered to microtubules. We demonstrate the use of the HyPer-Tau sensor for three applications; dose-dependent response of human cells to exogenous hydrogen peroxide, a model immune response of mouse macrophages to stimulation by bacterial toxin, and a spatially-resolved response to localized delivery of hydrogen peroxide. These results demonstrate that HyPer-Tau can be used as an effective tool for tracking changes in spatially localized intracellular hydrogen peroxide and for future applications in redox signaling.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Cell Line
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / metabolism*
  • Hydrogen Peroxide / pharmacology
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism*
  • Lipopolysaccharides / pharmacology
  • Macrophages / cytology
  • Macrophages / drug effects
  • Macrophages / metabolism*
  • Mice
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Oxidants / metabolism
  • Oxidants / pharmacology
  • Oxidation-Reduction
  • Plasmids / genetics
  • Plasmids / metabolism
  • Time-Lapse Imaging / methods
  • tau Proteins / genetics
  • tau Proteins / metabolism

Substances

  • Lipopolysaccharides
  • Oxidants
  • tau Proteins
  • Hydrogen Peroxide