H2O2: A Chemoattractant?
- Authors
- Enyedi, B., and Niethammer, P.
- ID
- ZDB-PUB-130722-13
- Date
- 2013
- Source
- Methods in enzymology 528: 237-255 (Chapter)
- Registered Authors
- Enyedi, Balázs, Niethammer, Philipp
- Keywords
- H2O2, chemotaxis, HyPer, leukocyte, zebrafish, wound, DUOX
- 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
- PubMed
- 23849869 Full text @ Methods Enzymol.
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.