title: H2O2: a chemoattractant?
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.
authors.affiliation: Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA.
authors.forename: Balázs
authors.initials: B
authors.lastname: Enyedi
authors.forename: Philipp
authors.initials: P
authors.lastname: Niethammer
mesh_terms: D058500:Animal Fins / Q000187:drug effects* / Q000276:immunology / Q000293:injuries; D000818:Animals; D015374:Biosensing Techniques; D002630:Chemotactic Factors / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D017930:Genes, Reporter; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D007113:Immunity, Innate; D007249:Inflammation; D007814:Larva / Q000166:cytology / Q000276:immunology; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D018613:Microscopy, Confocal; D010957:Plasmids; D014947:Wounds and Injuries / Q000276:immunology / Q000473:pathology; D015027:Zebrafish / Q000276:immunology / Q000293:injuries
chemical_list: D002630:Chemotactic Factors; D006861:Hydrogen Peroxide
keywords: Chemotaxis; DUOX; H(2)O(2); HyPer; Leukocyte; Wound; Zebrafish