ATP modulates acute inflammation in vivo through dual oxidase 1-derived H2O2 production and NF-κB activation.
Dual oxidase 1 (Duox1) is the NADPH oxidase responsible for the H2O2 gradient formed in tissues after injury to trigger the early recruitment of leukocytes. Little is known about the signals that modulate H2O2 release from DUOX1 and whether the H2O2 gradient can orchestrate the inflammatory response in vivo. In this study, we report on a dominant-negative form of zebrafish Duox1 that is able to inhibit endogenous Duox1 activity, H2O2 release and leukocyte recruitment after tissue injury, with none of the side effects associated with morpholino-mediated Duox1 knockdown. Using this specific tool, we found that ATP release following tissue injury activates purinergic P2Y receptors, and modulates Duox1 activity through phospholipase C (PLC) and intracellular calcium signaling in vivo. Furthermore, Duox1-derived H2O2 is able to trigger the NF-κB inflammatory signaling pathway. These data reveal that extracellular ATP acting as an early danger signal is responsible for the activation of Duox1 via a P2YR/PLC/Ca(2+) signaling pathway and the production of H2O2, which, in turn, is able to modulate in vivo not only the early recruitment of leukocytes to the wound but also the inflammatory response through activation of the NF-κB signaling pathway.
Laboratório de Carlota Saldanha, Instituto de Medicina Molecular, 1649-028 Lisbon, Portugal; Instituto de Bioquímica, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisbon, Portugal; Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain;
Sofia
S
de Oliveira
Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain; Instituto Murciano de Investigación Biosanitaria, 30120 Murcia, Spain; and.
Azucena
A
López-Muñoz
Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain; Instituto Murciano de Investigación Biosanitaria, 30120 Murcia, Spain; and.
Sergio
S
Candel
Instituto Murciano de Investigación Biosanitaria, 30120 Murcia, Spain; and Unidad de Inflamación y Cirugía Experimental, Centro de Investigación Biomédica en Red en el Área Temática de Enfermedades Hepáticas y Digestivas, Hospital Universitario Virgen de la Hospital Universitario Virgen de la Arrixaca, 30120 Murcia, Spain.
Pablo
P
Pelegrín
Laboratório de Carlota Saldanha, Instituto de Medicina Molecular, 1649-028 Lisbon, Portugal; Instituto de Bioquímica, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisbon, Portugal;
Ângelo
Â
Calado
Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain; Instituto Murciano de Investigación Biosanitaria, 30120 Murcia, Spain; and vmulero@um.es.
Victoriano
V
Mulero
D000208:Acute Disease; D000255:Adenosine Triphosphate / Q000276:immunology*; D000818:Animals; D020013:Calcium Signaling / Q000276:immunology*; D006861:Hydrogen Peroxide / Q000276:immunology*; D007249:Inflammation; D019255:NADPH Oxidases / Q000276:immunology*; D016328:NF-kappa B / Q000276:immunology*; D058470:Receptors, Purinergic P2Y / Q000276:immunology; D014947:Wounds and Injuries / Q000276:immunology*; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000276:immunology*
D016328:NF-kappa B; D058470:Receptors, Purinergic P2Y; D029961:Zebrafish Proteins; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases