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Published on: July 17, 2016
P2X(7) receptors on microglial cells mediate injury to cortical neurons in vitro
Stephen D Skaper1, Laura Facci, Ainsley A Culbert
1Neurology and GI Centre of Excellence for Drug Discovery, GlaxoSmithKline Research and Development Limited, New Frontiers Science Park, Third Avenue, Harlow CM19 5AW, Essex, United Kingdom. Stephen_D_Skaper@gsk.com
Abstract:
The P2X(7) receptor has been implicated in the release of cytokines and in the induction of cell death, and is up-regulated in a transgenic mouse model of Alzheimer's disease. Using cocultures of rat cortical neurons and microglia, we show that ATP and the more potent P2X(7) agonist benzoylbenzoyl-ATP (BzATP) cause neuronal cell injury. The deleterious effects of BzATP-treated microglia were prevented by nonselective P2X antagonists (PPADS and oxidized ATP) and by the more selective P2X(7) antagonist Brilliant Blue G. Similar concentrations of BzATP caused release of superoxide and nitric oxide from isolated microglia, and neuronal cell injury was attenuated by a superoxide dismutase mimetic and by a peroxynitrite decomposition catalyst, suggesting a role for reactive oxide species. Cocultures composed of wild-type cortical neurons, and microglia from P2X(7) receptor-deficient mice failed to exhibit neuronal cell injury in the presence of BzATP, but retained sensitivity to injury when microglia were derived from genotypically matched normal (P2X(7) (+/+) mice), thereby establishing P2X(7) involvement in the injury process. P2X(7) receptor activation on microglia thus appears necessary for microglial-mediated injury of neurons, and proposes that targeting P2X(7) receptors may constitute a novel approach for the treatment of acute and chronic neurodegenerative disorders where a microglial component is evident.
Insights
Activation of the P2X(7) receptor on microglia causes neuronal cell injury, suggesting it as a therapeutic target for neurodegenerative diseases. This study demonstrates P2X(7) receptor involvement in microglial-mediated neuronal damage.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- The P2X(7) receptor is involved in cytokine release and cell death.
- P2X(7) receptor is upregulated in Alzheimer's disease models.
- Microglia play a role in neuroinflammation and neurodegeneration.
Purpose of the Study:
- To investigate the role of the P2X(7) receptor in microglial-mediated neuronal injury.
- To determine if P2X(7) receptor activation on microglia leads to neuronal cell damage.
- To explore P2X(7) receptor antagonists as potential therapeutics for neurodegenerative disorders.
Main Methods:
- Coculture of rat cortical neurons and microglia.
- Treatment with ATP and benzoylbenzoyl-ATP (BzATP), a P2X(7) agonist.
- Use of P2X(7) receptor antagonists (PPADS, oxidized ATP, Brilliant Blue G).
- Assessment of neuronal cell injury and reactive oxygen species (ROS) release.
- Experiments using microglia from P2X(7) receptor-deficient mice.
Main Results:
- BzATP induced neuronal cell injury in cocultures.
- P2X(7) receptor antagonists prevented BzATP-induced neuronal injury.
- BzATP stimulated superoxide and nitric oxide release from microglia.
- Neuronal injury was attenuated by antioxidants, indicating a role for ROS.
- Microglia from P2X(7) receptor-deficient mice did not cause neuronal injury upon BzATP stimulation.
Conclusions:
- P2X(7) receptor activation on microglia is necessary for microglial-mediated neuronal injury.
- Reactive oxygen species are involved in the P2X(7) receptor-mediated neuronal damage.
- Targeting P2X(7) receptors may offer a novel therapeutic strategy for neurodegenerative diseases with a microglial component.
