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α-Synuclein Protects Neurons from Apoptosis Downstream of Free-Radical Production Through Modulation of the MAPK Signalling Pathway

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Abstract

α-Synuclein is a pre-synaptic chaperone and its accumulation contributes to differential cell loss in Parkinson’s disease. Cytoplasmic expression of α-synuclein can directly modulate apoptotic pathways and contribute to cell survival, whereas induced over-expression of the protein causes oxidative stress through mitochondrial and cytosolic free-radical production. This study aimed to clarify the contribution of endogenous α-synuclein to oxidative stress and its association with cell death. Primary cortical neurons were derived from α-synuclein knock-out (Snca-/-) and wild-type (C57BL/6; WT) mice and treated with in vitro models of oxidative-stress, complex I inhibition and excitotoxicity. Mitochondrial free radical production was determined in isolated mitochondria derived from each mouse strain. Snca-/- derived cortical cultures were more susceptible (P < 0.05) to oxidative-stress, but not excitotoxicity. This result was determined by significant increases in cell death (Propidium-Iodide staining) after 6 h treatment in Snca-/- (45 % ± 2.7 SEM), relative to WT (33 % ± 3.9 SEM) cultures. α-Synuclein also confers significant (P < 0.05) resistance to low-dose (5 nM) rotenone toxicity, with a twofold reduction in cell death in WT, compared with Snca-/- cortical neurons. The expression of α-synuclein had no effect on cortical glutathione levels, or the production of reactive oxygen intermediates in isolated mitochondria. These data indicate that endogenous levels of α-synuclein confer resistance to oxidative stress downstream of free radical production and scavenging. The current data suggest that α-synuclein prevents cytochrome c release and apoptosis through inhibition of the MAPK signalling pathway.

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Acknowledgments

We would like to acknowledge Dr Rob Gasperini for his assistance with mitochondrial isolation and function assays. This study was funded by National Health and Medical Research Council (NHMRC), NHMRC Career Development Award and Select Foundation Fellowship to TD, Wicking Dementia Research and Education Centre, Motor Neuron Disease Research Institute of Australia Bill Gole Fellowship and Alzheimer’s Association of Australia Fellowship to AK. Laser Scanning microscopy was performed on a Leica LSM 510, provided by the Central Science Laboratory (CSL) of the University of Tasmania.

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  1. Menzies Research Institute Tasmania, University of Tasmania, Private Bag 23, Hobart, TAS, 7000, Australia

    Ruth E. J. Musgrove, Anna E. King & Tracey C. Dickson

  2. Wicking Dementia Research and Education Centre, University of Tasmania, Hobart, TAS, 7000, Australia

    Anna E. King

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  1. Ruth E. J. Musgrove
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  2. Anna E. King
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Correspondence to Tracey C. Dickson.

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Musgrove, R.E.J., King, A.E. & Dickson, T.C. α-Synuclein Protects Neurons from Apoptosis Downstream of Free-Radical Production Through Modulation of the MAPK Signalling Pathway. Neurotox Res 23, 358–369 (2013). https://doi.org/10.1007/s12640-012-9352-5

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  • DOI: https://doi.org/10.1007/s12640-012-9352-5

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