H2O2: A Chemoattractant?

Methods Enzymol. 2013;528:237-55. doi: 10.1016/B978-0-12-405881-1.00014-8.

Abstract

H2O2 is a relatively stable, rapidly diffusing reactive oxygen species that has been recently implicated as a mediator of leukocyte recruitment to epithelial wounds and transformed cells in zebrafish. Whether H2O2 activates the innate immune response by acting as a bona fide chemoattractant, enhancing chemoattractant sensing, or triggering production of other chemoattractive ligands remains largely unclear. Here, we describe the basic experimental procedures required to study these questions. We present a detailed protocol of the zebrafish tail fin wounding assay and explain how to use it for analyzing leukocyte chemotaxis in vivo. We further outline a method for H2O2 measurement in live zebrafish larvae using the genetically encoded sensor HyPer on a wide-field and a spinning disk confocal microscope. These methods provide a basis for dissecting the role of H2O2 in leukocyte chemotaxis in a vertebrate animal.

Keywords: Chemotaxis; DUOX; H(2)O(2); HyPer; Leukocyte; Wound; Zebrafish.

Publication types

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

MeSH terms

  • Animal Fins / drug effects*
  • Animal Fins / immunology
  • Animal Fins / injuries
  • Animals
  • Biosensing Techniques*
  • Chemotactic Factors / analysis
  • Chemotactic Factors / metabolism*
  • Chemotactic Factors / pharmacology
  • Chemotaxis, Leukocyte / drug effects*
  • Genes, Reporter
  • Hydrogen Peroxide / analysis
  • Hydrogen Peroxide / metabolism*
  • Hydrogen Peroxide / pharmacology
  • Image Processing, Computer-Assisted
  • Immunity, Innate
  • Inflammation
  • Larva / cytology
  • Larva / immunology
  • Leukocytes / cytology
  • Leukocytes / drug effects*
  • Leukocytes / immunology
  • Microscopy, Confocal
  • Plasmids
  • Wounds and Injuries / immunology
  • Wounds and Injuries / pathology
  • Zebrafish / immunology
  • Zebrafish / injuries

Substances

  • Chemotactic Factors
  • Hydrogen Peroxide