Original ContributionNucleotide receptor signaling in murine macrophages is linked to reactive oxygen species generation
Section snippets
Reagents
The nucleotides BzATP, ATP, UTP, α, β-methylene-ATP were obtained from Sigma (St. Louis, MO, USA). Lipopolysaccharide (Escherichia coli, serotype 0111:B4), phorbol 12-myristate 13-acetate (PMA), anisomycin, N-acetylcysteine (NAC), ascorbic acid, and methylthiazole tetrazolium (MTT) were also purchased from Sigma. The ROS indicator 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) was obtained from Molecular Probes (Eugene, OR, USA). The cell-permeable peptide JNK inhibitor (L)-JNKI1 and JNK
Extracellular nucleotide-stimulated ROS production
Although P2-receptor-stimulated ROS production has been reported for some cell types such as neutrophils, the participation of P2X7 in nucleotide-dependent ROS production in macrophages is less clear [23], [52]. Because the production of ROS is important for various macrophage functions including bacterial killing, we first sought to establish whether P2X7 agonists can stimulate the production of ROS in murine RAW 264.7 macrophages. In this regard, treatment of these cells with the selective P2X
Discussion
High levels of extracellular nucleotides are present at sites of inflammation, platelet degranulation, and cellular damage/lysis [59]. These extracellular nucleotides activate receptors on various immune cells including monocytes and macrophages [1], [54]. This activation process results in the increased production and release of multiple inflammatory mediators, and in the current study we provide evidence that the nucleotide receptor P2X7 can promote the generation of ROS in murine
Acknowledgments
This work was supported by National Institutes of Health Grants HL56396 and AI50500. Z.A.P. was supported by Biotechnology Training Program NIH 5 T32 GM08349, University of Wisconsin. A.N.G. was supported by Hematology Training Program NIH 5 T32 HL07899, University of Wisconsin.
References (64)
- et al.
Nucleotide receptors: an emerging family of regulatory molecules in blood cells
Blood
(2001) The P2Z purinoceptor: an intriguing role in immunity, inflammation and cell death
Immunol. Today
(1995)- et al.
Altered cytokine production in mice lacking P2X7 receptors
J. Biol. Chem.
(2001) - et al.
ATP-induced killing of mycobacteria by human macrophages is mediated by purinergic P2Z (P2X7) receptors
Immunity
(1997) - et al.
Extracellular ATP triggers superoxide production in human neutrophils
Biochem. Biophys. Res. Commun.
(1989) - et al.
Adenosine triphosphate-induced oxygen radical production and CD11b up-regulation: Ca++ mobilization and actin reorganization in human eosinophils
Blood
(2000) - et al.
P2 purinergic receptors of human eosinophils: characterization and coupling to oxygen radical production
FEBS Lett.
(2000) - et al.
P2X7 mediates superoxide production in primary microglia and is up-regulated in a transgenic mouse model of Alzheimer's disease
J. Biol. Chem.
(2003) - et al.
Inside the neutrophil phagosome: oxidants, myeloperoxidase, and bacterial killing
Blood
(1998) - et al.
Activation of NF-kappa B by the respiratory burst of macrophages
Free Radic. Biol. Med.
(1996)
Activation of mitogen-activated protein kinase by H2O2: role in cell survival following oxidant injury
J. Biol. Chem.
Reactive oxygen species mediate cytokine activation of c-Jun NH2-terminal kinases
J. Biol. Chem.
AP-1 activation through endogenous H2O2 generation by alveolar macrophages
Free Radic. Biol. Med.
Regulation and measurement of oxidative stress in apoptosis
J. Immunol. Methods
p21ras as a common signaling target of reactive free radicals and cellular redox stress
J. Biol. Chem.
Inhibition of PTPs by H2O2 regulates the activation of distinct MAPK pathways
Free Radic. Biol. Med.
Signal transduction by the c-Jun N-terminal kinase (JNK)—from inflammation to development
Curr. Opin. Cell Biol.
AP-1 function and regulation
Curr. Opin. Cell Biol.
Stress-activated protein kinase/JNK activation and apoptotic induction by the macrophage P2X7 nucleotide receptor
J. Biol. Chem.
Signaling through P2X7 receptor in human T cells involves p56lck, MAP kinases, and transcription factors AP-1 and NF-kappa B
J. Biol. Chem.
Molecular basis of interferon-γ and lipopolysaccharide enhancement of phagocyte respiratory burst capability: studies on the gene expression of several NADPH oxidase components
J. Biol. Chem.
The antioxidant action of N-acetylcysteine: its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid
Free Radic. Biol. Med.
Purinergic receptor modulation of lipopolysaccharide signaling and inducible nitric-oxide synthase expression in RAW 264.7 macrophages
J. Biol. Chem.
Molecular physiology of P2X receptors
Physiol. Rev.
Leukocyte P2 receptors: a novel target for anti-inflammatory and antitumor therapy
Curr. Drug Targets Cardiovasc. Haematol. Disord.
Extracellular ATP triggers IL-1 beta release by activating the purinergic P2Z receptor of human macrophages
J. Immunol.
P2X7 nucleotide receptor: modulation of LPS-induced macrophage signaling and mediator production
Drug Dev. Res.
Proteomic and functional evidence for a P2X7 receptor signalling complex
EMBO J.
Cutting edge: the nucleotide receptor P2X7 contains multiple protein- and lipid-interaction motifs including a potential binding site for bacterial lipopolysaccharide
J. Immunol.
The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7)
Science
Platelet enhancement of O2− responses in stimulated human neutrophils: identification of platelet factor as adenine nucleotide
Lab. Invest.
Regulatory effects of adenosine and adenine nucleotides on oxygen radical responses of neutrophils
Lab. Invest.
Cited by (68)
Host P2X<inf>7</inf>R-p<inf>38</inf>MAPK axis mediated intra-macrophage leishmanicidal activity of Spergulin-A
2022, Experimental ParasitologyThe role of NADPH oxidases in infectious and inflammatory diseases
2021, Redox BiologyPurinergic signaling in the modulation of redox biology
2021, Redox BiologyHow the phagocyte NADPH oxidase regulates innate immunity
2018, Free Radical Biology and MedicineCitation Excerpt :Furthermore, there are definitely situations in which NLRP3 activation potentiates ROS production. Treatment of macrophages with ATP results in production of reactive oxygen species, which stimulate the PI3 kinase pathways and contribute to Akt and ERK1/2 activation [11,58,66]. Conversely, not only does the P2X7-mediated activation of the NLRP3 inflammasome by ATP drive the production of reactive oxygen species but the ROS-driven effects on PI3 kinase are necessary for the activation of caspase 1 and then IL-1β and IL-18 [11].
The phagocyte respiratory burst: Historical perspectives and recent advances
2017, Immunology Letters
- 1
Current address: Department of Chemistry, Lawrence University, Appleton, WI 54912, USA.