Abstract
Parkinson’s disease (PD) and dementia with Lewy bodies (DLB) are characterized by intracellular inclusions composed mostly of α-synuclein (Baba et al., Am J Pathol 152:879–884, 1998). How inclusion formation impacts neuronal function prior to death is key to understanding disease progression and identifying therapeutic windows. In the α-synuclein fibril model, exposure of primary neurons to α-synuclein fibrils induces endogenously expressed α-synuclein to form inclusions which closely resembles pathologic mechanisms in humans with PD and DLB (Volpicelli-Daley et al., Neuron 72, 57–71, 2011). In this model, at 7 days after exposure of neurons to fibrils, when there is no neuron death, inclusions in the axon selectively impair axonal transport of endosomes carrying the TrkB receptor and LC3-positive autophagosomes (Volpicelli-Daley et al., Mol Biol Cell 25:4010–4023, 2014). In addition, the frequency and amplitude of spontaneous Ca2+ transients are reduced in neurons 7 days after fibril exposure. Here we discuss protocols for plating primary hippocampal neurons, generating fibrils and measuring axonal transport and Ca2+ transients. These assays provide additional assays of neurotoxicity allowing researchers to determine if a therapeutic intervention can prevent neuronal defects before intractable neurodegeneration.
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References
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Volpicelli-Daley, L.A. (2019). Assays for Neuronal Defects Caused by Early Formation of α-Synuclein Inclusions in Primary Cultured Neurons. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_1
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DOI: https://doi.org/10.1007/978-1-4939-9124-2_1
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