Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Journal of Neuroinflammation
Published in: Journal of Neuroinflammation 1/2010

Open Access 01-12-2010 | Research

Extracellular ATP and the P2X7receptor in astrocyte-mediated motor neuron death: implications for amyotrophic lateral sclerosis

Authors: Mandi Gandelman, Hugo Peluffo, Joseph S Beckman, Patricia Cassina, Luis Barbeito

Published in: Journal of Neuroinflammation | Issue 1/2010

Login to get access

Abstract

Background

During pathology of the nervous system, increased extracellular ATP acts both as a cytotoxic factor and pro-inflammatory mediator through P2X7 receptors. In animal models of amyotrophic lateral sclerosis (ALS), astrocytes expressing superoxide dismutase 1 (SOD1G93A) mutations display a neuroinflammatory phenotype and contribute to disease progression and motor neuron death. Here we studied the role of extracellular ATP acting through P2X7 receptors as an initiator of a neurotoxic phenotype that leads to astrocyte-mediated motor neuron death in non-transgenic and SOD1G93A astrocytes.

Methods

We evaluated motor neuron survival after co-culture with SOD1G93A or non-transgenic astrocytes pretreated with agents known to modulate ATP release or P2X7 receptor. We also characterized astrocyte proliferation and extracellular ATP degradation.

Results

Repeated stimulation by ATP or the P2X7-selective agonist BzATP caused astrocytes to become neurotoxic, inducing death of motor neurons. Involvement of P2X7 receptor was further confirmed by Brilliant blue G inhibition of ATP and BzATP effects. In SOD1G93A astrocyte cultures, pharmacological inhibition of P2X7 receptor or increased extracellular ATP degradation with the enzyme apyrase was sufficient to completely abolish their toxicity towards motor neurons. SOD1G93A astrocytes also displayed increased ATP-dependent proliferation and a basal increase in extracellular ATP degradation.

Conclusions

Here we found that P2X7 receptor activation in spinal cord astrocytes initiated a neurotoxic phenotype that leads to motor neuron death. Remarkably, the neurotoxic phenotype of SOD1G93A astrocytes depended upon basal activation the P2X7 receptor. Thus, pharmacological inhibition of P2X7 receptor might reduce neuroinflammation in ALS through astrocytes.
Appendix
Available only for authorised users
Literature
1.
Rowland LP, Shneider NA: Amyotrophic lateral sclerosis. N Engl J Med. 2001, 344: 1688-1700. 10.1056/NEJM200105313442207.CrossRefPubMed
2.
Rosen DR: Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature. 1993, 364: 362.PubMed
3.
Beckman JS, Estevez AG, Crow JP, Barbeito L: Superoxide dismutase and the death of motoneurons in ALS. Trends Neurosci. 2001, 24: S15-20. 10.1016/S0166-2236(00)01981-0.CrossRefPubMed
4.
Harraz MM, Marden JJ, Zhou W, Zhang Y, Williams A, Sharov VS, Nelson K, Luo M, Paulson H, Schoneich C, Engelhardt JF: SOD1 mutations disrupt redox-sensitive Rac regulation of NADPH oxidase in a familial ALS model. J Clin Invest. 2008, 118: 659-670.PubMedCentralPubMed
5.
Papadimitriou D, Le Verche V, Jacquier A, Ikiz B, Przedborski S, Re DB: Inflammation in ALS and SMA: Sorting out the good from the evil. Neurobiol Dis. 2009
6.
Kikuchi H, Almer G, Yamashita S, Guegan C, Nagai M, Xu Z, Sosunov AA, McKhann GM, Przedborski S: Spinal cord endoplasmic reticulum stress associated with a microsomal accumulation of mutant superoxide dismutase-1 in an ALS model. Proc Natl Acad Sci USA. 2006, 103: 6025-6030. 10.1073/pnas.0509227103.PubMedCentralCrossRefPubMed
7.
Rothstein JD, Tsai G, Kuncl RW, Clawson L, Cornblath DR, Drachman DB, Pestronk A, Stauch BL, Coyle JT: Abnormal excitatory amino acid metabolism in amyotrophic lateral sclerosis. Ann Neurol. 1990, 28: 18-25. 10.1002/ana.410280106.CrossRefPubMed
8.
Wong PC, Pardo CA, Borchelt DR, Lee MK, Copeland NG, Jenkins NA, Sisodia SS, Cleveland DW, Price DL: An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria. Neuron. 1995, 14: 1105-1116. 10.1016/0896-6273(95)90259-7.CrossRefPubMed
9.
Bruijn LI, Houseweart MK, Kato S, Anderson KL, Anderson SD, Ohama E, Reaume AG, Scott RW, Cleveland DW: Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1. Science. 1998, 281: 1851-1854. 10.1126/science.281.5384.1851.CrossRefPubMed
10.
Ilieva H, Polymenidou M, Cleveland DW: Non-cell autonomous toxicity in neurodegenerative disorders: ALS and beyond. J Cell Biol. 2009, 187: 761-72. 10.1083/jcb.200908164.PubMedCentralCrossRefPubMed
11.
McGeer PL, McGeer EG: Inflammation and the degenerative diseases of aging. Ann N Y Acad Sci. 2004, 1035: 104-116. 10.1196/annals.1332.007.CrossRefPubMed
12.
Barbeito LH, Pehar M, Cassina P, Vargas MR, Peluffo H, Viera L, Estevez AG, Beckman JS: A role for astrocytes in motor neuron loss in amyotrophic lateral sclerosis. Brain Res Brain Res Rev. 2004, 47: 263-274. 10.1016/j.brainresrev.2004.05.003.CrossRefPubMed
13.
Bruijn LI, Miller TM, Cleveland DW: Unraveling the mechanisms involved in motor neuron degeneration in ALS. Annu Rev Neurosci. 2004, 27: 723-749. 10.1146/annurev.neuro.27.070203.144244.CrossRefPubMed
14.
Vargas MR, Pehar M, Cassina P, Beckman JS, Barbeito L: Increased glutathione biosynthesis by Nrf2 activation in astrocytes prevents p75NTR-dependent motor neuron apoptosis. J Neurochem. 2006, 97: 687-696. 10.1111/j.1471-4159.2006.03742.x.CrossRefPubMed
15.
Di Giorgio FP, Boulting GL, Bobrowicz S, Eggan KC: Human embryonic stem cell-derived motor neurons are sensitive to the toxic effect of glial cells carrying an ALS-causing mutation. Cell Stem Cell. 2008, 3: 637-648. 10.1016/j.stem.2008.09.017.CrossRefPubMed
16.
Di Giorgio FP, Carrasco MA, Siao MC, Maniatis T, Eggan K: Non-cell autonomous effect of glia on motor neurons in an embryonic stem cell-based ALS model. Nat Neurosci. 2007, 10: 608-614. 10.1038/nn1885.PubMedCentralCrossRefPubMed
17.
Marchetto MC, Muotri AR, Mu Y, Smith AM, Cezar GG, Gage FH: Non-cell-autonomous effect of human SOD1 G37R astrocytes on motor neurons derived from human embryonic stem cells. Cell Stem Cell. 2008, 3: 649-657. 10.1016/j.stem.2008.10.001.CrossRefPubMed
18.
Nagai M, Re DB, Nagata T, Chalazonitis A, Jessell TM, Wichterle H, Przedborski S: Astrocytes expressing ALS-linked mutated SOD1 release factors selectively toxic to motor neurons. Nat Neurosci. 2007, 10: 615-622. 10.1038/nn1876.PubMedCentralCrossRefPubMed
19.
Cassina P, Cassina A, Pehar M, Castellanos R, Gandelman M, de Leon A, Robinson KM, Mason RP, Beckman JS, Barbeito L, Radi R: Mitochondrial dysfunction in SOD1G93A-bearing astrocytes promotes motor neuron degeneration: prevention by mitochondrial-targeted antioxidants. J Neurosci. 2008, 28: 4115-4122. 10.1523/JNEUROSCI.5308-07.2008.PubMedCentralCrossRefPubMed
20.
Cassina P, Pehar M, Vargas MR, Castellanos R, Barbeito AG, Estevez AG, Thompson JA, Beckman JS, Barbeito L: Astrocyte activation by fibroblast growth factor-1 and motor neuron apoptosis: implications for amyotrophic lateral sclerosis. J Neurochem. 2005, 93: 38-46. 10.1111/j.1471-4159.2004.02984.x.CrossRefPubMed
21.
Vargas MR, Pehar M, Cassina P, Martinez-Palma L, Thompson JA, Beckman JS, Barbeito L: Fibroblast growth factor-1 induces heme oxygenase-1 via nuclear factor erythroid 2-related factor 2 (Nrf2) in spinal cord astrocytes: consequences for motor neuron survival. J Biol Chem. 2005, 280: 25571-25579. 10.1074/jbc.M501920200.CrossRefPubMed
22.
Hensley K, Abdel-Moaty H, Hunter J, Mhatre M, Mou S, Nguyen K, Potapova T, Pye QN, Qi M, Rice H, Stewart C, Stroukoff K, West M: Primary glia expressing the G93A-SOD1 mutation present a neuroinflammatory phenotype and provide a cellular system for studies of glial inflammation. J Neuroinflammation. 2006, 3: 2-10.1186/1742-2094-3-2.PubMedCentralCrossRefPubMed
23.
Pehar M, Vargas MR, Robinson KM, Cassina P, England P, Beckman JS, Alzari PM, Barbeito L: Peroxynitrite transforms nerve growth factor into an apoptotic factor for motor neurons. Free Radic Biol Med. 2006, 41: 1632-1644. 10.1016/j.freeradbiomed.2006.08.010.CrossRefPubMed
24.
Burnstock G: Purinergic signalling and disorders of the central nervous system. Nat Rev Drug Discov. 2008, 7: 575-590. 10.1038/nrd2605.CrossRefPubMed
25.
Wang X, Arcuino G, Takano T, Lin J, Peng WG, Wan P, Li P, Xu Q, Liu QS, Goldman SA, Nedergaard M: P2X7 receptor inhibition improves recovery after spinal cord injury. Nat Med. 2004, 10: 821-827. 10.1038/nm1082.CrossRefPubMed
26.
Phillis JW, O'Regan MH, Perkins LM: Adenosine 5'-triphosphate release from the normoxic and hypoxic in vivo rat cerebral cortex. Neurosci Lett. 1993, 151: 94-96. 10.1016/0304-3940(93)90054-O.CrossRefPubMed
27.
Melani A, Turchi D, Vannucchi MG, Cipriani S, Gianfriddo M, Pedata F: ATP extracellular concentrations are increased in the rat striatum during in vivo ischemia. Neurochem Int. 2005, 47: 442-448. 10.1016/j.neuint.2005.05.014.CrossRefPubMed
28.
Piccini A, Carta S, Tassi S, Lasiglie D, Fossati G, Rubartelli A: ATP is released by monocytes stimulated with pathogen-sensing receptor ligands and induces IL-1beta and IL-18 secretion in an autocrine way. Proc Natl Acad Sci USA. 2008, 105: 8067-8072. 10.1073/pnas.0709684105.PubMedCentralCrossRefPubMed
29.
30.
Franke H, Gunther A, Grosche J, Schmidt R, Rossner S, Reinhardt R, Faber-Zuschratter H, Schneider D, Illes P: P2X7 receptor expression after ischemia in the cerebral cortex of rats. J Neuropathol Exp Neurol. 2004, 63: 686-699.PubMed
31.
Lovatt D, Sonnewald U, Waagepetersen HS, Schousboe A, He W, Lin JH, Han X, Takano T, Wang S, Sim FJ, Goldman SA, Nedergaard M: The transcriptome and metabolic gene signature of protoplasmic astrocytes in the adult murine cortex. J Neurosci. 2007, 27: 12255-12266. 10.1523/JNEUROSCI.3404-07.2007.CrossRefPubMed
32.
Narcisse L, Scemes E, Zhao Y, Lee SC, Brosnan CF: The cytokine IL-1beta transiently enhances P2X7 receptor expression and function in human astrocytes. Glia. 2005, 49: 245-258. 10.1002/glia.20110.PubMedCentralCrossRefPubMed
33.
John GR, Simpson JE, Woodroofe MN, Lee SC, Brosnan CF: Extracellular nucleotides differentially regulate interleukin-1beta signaling in primary human astrocytes: implications for inflammatory gene expression. J Neurosci. 2001, 21: 4134-4142.PubMed
34.
Panenka W, Jijon H, Herx LM, Armstrong JN, Feighan D, Wei T, Yong VW, Ransohoff RM, MacVicar BA: P2X7-like receptor activation in astrocytes increases chemokine monocyte chemoattractant protein-1 expression via mitogen-activated protein kinase. J Neurosci. 2001, 21: 7135-7142.PubMed
35.
Ryu JK, McLarnon JG: Block of purinergic P2X(7) receptor is neuroprotective in an animal model of Alzheimer's disease. Neuroreport. 2008, 19: 1715-1719. 10.1097/WNR.0b013e3283179333.CrossRefPubMed
36.
Matute C, Torre I, Perez-Cerda F, Perez-Samartin A, Alberdi E, Etxebarria E, Arranz AM, Ravid R, Rodriguez-Antiguedad A, Sanchez-Gomez M, Domercq M: P2X(7) receptor blockade prevents ATP excitotoxicity in oligodendrocytes and ameliorates experimental autoimmune encephalomyelitis. J Neurosci. 2007, 27: 9525-9533. 10.1523/JNEUROSCI.0579-07.2007.CrossRefPubMed
37.
Diaz-Hernandez M, Diez-Zaera M, Sanchez-Nogueiro J, Gomez-Villafuertes R, Canals JM, Alberch J, Miras-Portugal MT, Lucas JJ: Altered P2X7-receptor level and function in mouse models of Huntington's disease and therapeutic efficacy of antagonist administration. FASEB J. 2009, 23: 1893-1906. 10.1096/fj.08-122275.CrossRefPubMed
38.
Peng W, Cotrina ML, Han X, Yu H, Bekar L, Blum L, Takano T, Tian GF, Goldman SA, Nedergaard M: Systemic administration of an antagonist of the ATP-sensitive receptor P2X7 improves recovery after spinal cord injury. Proc Natl Acad Sci USA. 2009, 106: 12489-12493. 10.1073/pnas.0902531106.PubMedCentralCrossRefPubMed
39.
Yiangou Y, Facer P, Durrenberger P, Chessell IP, Naylor A, Bountra C, Banati RR, Anand P: COX-2, CB2 and P2X7-immunoreactivities are increased in activated microglial cells/macrophages of multiple sclerosis and amyotrophic lateral sclerosis spinal cord. BMC Neurol. 2006, 6: 12-10.1186/1471-2377-6-12.PubMedCentralCrossRefPubMed
40.
Casanovas A, Hernandez S, Tarabal O, Rossello J, Esquerda JE: Strong P2X4 purinergic receptor-like immunoreactivity is selectively associated with degenerating neurons in transgenic rodent models of amyotrophic lateral sclerosis. J Comp Neurol. 2008, 506: 75-92. 10.1002/cne.21527.CrossRefPubMed
41.
D'Ambrosi N, Finocchi P, Apolloni S, Cozzolino M, Ferri A, Padovano V, Pietrini G, Carri MT, Volonte C: The proinflammatory action of microglial P2 receptors is enhanced in SOD1 models for amyotrophic lateral sclerosis. J Immunol. 2009, 183: 4648-4656. 10.4049/jimmunol.0901212.CrossRefPubMed
42.
Cassina P, Peluffo H, Pehar M, Martinez-Palma L, Ressia A, Beckman JS, Estevez AG, Barbeito L: Peroxynitrite triggers a phenotypic transformation in spinal cord astrocytes that induces motor neuron apoptosis. J Neurosci Res. 2002, 67: 21-29. 10.1002/jnr.10107.CrossRefPubMed
43.
Henderson CE, Bloch-Gallego E, Camu W: Purification and culture of embryonic motor neurons. 1995, Oxford: IRL Press
44.
Estevez AG, Sahawneh MA, Lange PS, Bae N, Egea M, Ratan RR: Arginase 1 regulation of nitric oxide production is key to survival of trophic factor-deprived motor neurons. J Neurosci. 2006, 26: 8512-8516. 10.1523/JNEUROSCI.0728-06.2006.PubMedCentralCrossRefPubMed
45.
Wink MR, Braganhol E, Tamajusuku AS, Casali EA, Karl J, Barreto-Chaves ML, Sarkis JJ, Battastini AM: Extracellular adenine nucleotides metabolism in astrocyte cultures from different brain regions. Neurochem Int. 2003, 43: 621-628. 10.1016/S0197-0186(03)00094-9.CrossRefPubMed
46.
Teng RJ, Ye YZ, Parks DA, Beckman JS: Urate produced during hypoxia protects heart proteins from peroxynitrite-mediated protein nitration. Free Radic Biol Med. 2002, 33: 1243-1249. 10.1016/S0891-5849(02)01020-1.CrossRefPubMed
47.
Santos CX, Anjos EI, Augusto O: Uric acid oxidation by peroxynitrite: multiple reactions, free radical formation, and amplification of lipid oxidation. Arch Biochem Biophys. 1999, 372: 285-294. 10.1006/abbi.1999.1491.CrossRefPubMed
48.
Neary JT, Kang Y: Signaling from P2 nucleotide receptors to protein kinase cascades induced by CNS injury: implications for reactive gliosis and neurodegeneration. Mol Neurobiol. 2005, 31: 95-103. 10.1385/MN:31:1-3:095.CrossRefPubMed
49.
Rathbone MP, Middlemiss PJ, Kim JK, Gysbers JW, DeForge SP, Smith RW, Hughes DW: Adenosine and its nucleotides stimulate proliferation of chick astrocytes and human astrocytoma cells. Neurosci Res. 1992, 13: 1-17. 10.1016/0168-0102(92)90030-G.CrossRefPubMed
50.
Ciccarelli R, Di Iorio P, D'Alimonte I, Giuliani P, Florio T, Caciagli F, Middlemiss PJ, Rathbone MP: Cultured astrocyte proliferation induced by extracellular guanosine involves endogenous adenosine and is raised by the co-presence of microglia. Glia. 2000, 29: 202-211. 10.1002/(SICI)1098-1136(20000201)29:3<202::AID-GLIA2>3.0.CO;2-C.CrossRefPubMed
51.
Cotrina ML, Nedergaard M: Physiological and pathological functions of P2X7 receptor in the spinal cord. Purinergic Signal. 2009, 5: 223-232. 10.1007/s11302-009-9138-2.PubMedCentralCrossRefPubMed
52.
Surprenant A: Functional properties of native and cloned P2X receptors. Ciba Found Symp. 1996, 198: 208-219. discussion 219-222PubMed
53.
Bianchi BR, Lynch KJ, Touma E, Niforatos W, Burgard EC, Alexander KM, Park HS, Yu H, Metzger R, Kowaluk E, Jarvis MF, van Biesen T: Pharmacological characterization of recombinant human and rat P2X receptor subtypes. Eur J Pharmacol. 1999, 376: 127-138. 10.1016/S0014-2999(99)00350-7.CrossRefPubMed
54.
Skaper SD, Facci L, Culbert AA, Evans NA, Chessell I, Davis JB, Richardson JC: P2X(7) receptors on microglial cells mediate injury to cortical neurons in vitro. Glia. 2006, 54: 234-242. 10.1002/glia.20379.CrossRefPubMed
55.
Braun N, Zhu Y, Krieglstein J, Culmsee C, Zimmermann H: Upregulation of the enzyme chain hydrolyzing extracellular ATP after transient forebrain ischemia in the rat. J Neurosci. 1998, 18: 4891-4900.PubMed
56.
Nedeljkovic N, Bjelobaba I, Lavrnja I, Stojkov D, Pekovic S, Rakic L, Stojiljkovic M: Early temporal changes in ecto-nucleotidase activity after cortical stab injury in rat. Neurochem Res. 2008, 33: 873-879. 10.1007/s11064-007-9529-0.CrossRefPubMed
57.
Yamanaka K, Chun SJ, Boillee S, Fujimori-Tonou N, Yamashita H, Gutmann DH, Takahashi R, Misawa H, Cleveland DW: Astrocytes as determinants of disease progression in inherited amyotrophic lateral sclerosis. Nat Neurosci. 2008, 11: 251-253. 10.1038/nn2047.PubMedCentralCrossRefPubMed
Metadata
Title
Extracellular ATP and the P2X7receptor in astrocyte-mediated motor neuron death: implications for amyotrophic lateral sclerosis
Authors
Mandi Gandelman
Hugo Peluffo
Joseph S Beckman
Patricia Cassina
Luis Barbeito
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Journal of Neuroinflammation / Issue 1/2010
Electronic ISSN: 1742-2094
DOI
https://doi.org/10.1186/1742-2094-7-33

Other articles of this Issue 1/2010

Journal of Neuroinflammation 1/2010 Go to the issue

Research

Cerebral FDG-PET scanning abnormalities in optimally treated HIV patients

Review

Interleukin 18 in the CNS

Research

Cerebral ischemia induces microvascular pro-inflammatory cytokine expression via the MEK/ERK pathway

Short report

A role for spleen monocytes in post-ischemic brain inflammation and injury

Research

Hemin inhibits NO production by IL-1β-stimulated human astrocytes through induction of heme oxygenase-1 and reduction of p38 MAPK activation

Research

A neurotoxic regimen of methamphetamine exacerbates the febrile and neuroinflammatory response to a subsequent peripheral immune stimulus