JOURNAL OF
NEUROINFLAMMATION
Gandelman et al. Journal of Neuroinflammation 2010, 7:33
http://www.jneuroinflammation.com/content/7/1/33
Open Access RESEARCH
© 2010 Gandelman et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Com-
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Research
Extracellular ATP and the P2X
7
receptor in
astrocyte-mediated motor neuron death:
implications for amyotrophic lateral sclerosis
Mandi Gandelman
1,2
, Hugo Peluffo
1,2
, Joseph S Beckman
3
, Patricia Cassina
2
and Luis Barbeito*
1,4
Abstract
Background: During pathology of the nervous system, increased extracellular ATP acts both as a cytotoxic factor and
pro-inflammatory mediator through P2X
7
receptors. In animal models of amyotrophic lateral sclerosis (ALS), astrocytes
expressing superoxide dismutase 1 (SOD1
G93A
) mutations display a neuroinflammatory phenotype and contribute to
disease progression and motor neuron death. Here we studied the role of extracellular ATP acting through P2X
7
receptors as an initiator of a neurotoxic phenotype that leads to astrocyte-mediated motor neuron death in non-
transgenic and SOD1
G93A
astrocytes.
Methods: We evaluated motor neuron survival after co-culture with SOD1
G93A
or non-transgenic astrocytes pretreated
with agents known to modulate ATP release or P2X
7
receptor. We also characterized astrocyte proliferation and
extracellular ATP degradation.
Results: Repeated stimulation by ATP or the P2X
7
-selective agonist BzATP caused astrocytes to become neurotoxic,
inducing death of motor neurons. Involvement of P2X
7
receptor was further confirmed by Brilliant blue G inhibition of
ATP and BzATP effects. In SOD1
G93A
astrocyte cultures, pharmacological inhibition of P2X
7
receptor or increased
extracellular ATP degradation with the enzyme apyrase was sufficient to completely abolish their toxicity towards
motor neurons. SOD1
G93A
astrocytes also displayed increased ATP-dependent proliferation and a basal increase in
extracellular ATP degradation.
Conclusions: Here we found that P2X
7
receptor activation in spinal cord astrocytes initiated a neurotoxic phenotype
that leads to motor neuron death. Remarkably, the neurotoxic phenotype of SOD1
G93A
astrocytes depended upon
basal activation the P2X
7
receptor. Thus, pharmacological inhibition of P2X
7
receptor might reduce neuroinflammation
in ALS through astrocytes.
Background
Amyotrophic lateral sclerosis (ALS) is characterized by
the progressive degeneration of motor neurons in the spi-
nal cord, brainstem and motor cortex, leading to respira-
tory failure and death of affected patients within a few
years of diagnosis [1]. The discovery of mutations in the
gene encoding the antioxidant enzyme Cu/Zn superoxide
dismutase-1 (SOD1) in a subset of patients with familial
ALS has led to the development of transgenic animal
models expressing different SOD1 mutations [2]. These
animal models recapitulate the human disease, exhibiting
aberrant oxidative chemistry [3,4], neuroinflammation
[5], endoplasmic reticulum stress [6], glutamate excito-
toxicity [7], mitochondrial dysfunction [8] and protein
misfolding and aggregation [9]. However, the mecha-
nisms behind motor neuron death are unknown.
Accumulating evidence indicates that non-neuronal
cells contribute to motor neuron dysfunction and death
in ALS by the maintenance of a chronic inflammatory
response [10-12]. Activated microglia accumulate in the
spinal cord, producing inflammatory mediators and reac-
tive oxygen and nitrogen species [11]. Astrocytes, the
most abundant cells in the adult nervous system, also
* Correspondence: barbeito@pasteur.edu.uy
1
Neurodegeneration Laboratory, Institut Pasteur, Montevideo, Uruguay
Full list of author information is available at the end of the article