Oligomeric α-synuclein inhibits tubulin polymerization
Section snippets
Experimental procedures
Neurotoxicity induced by oligomeric α-synuclein. Oligomeric α-synuclein was produced using a modified protocol published by us recently [13]. Essentially, 100 μL of human recombinant α-synuclein (rPeptide, Athens, GA) was aged in vitro for 12 h with constant agitation using a magnetic stir bar at 37 °C. MES cells were cultured with the methods utilized routinely in our lab [14], and viability was tested with trypan blue (Invitrogen, Carlsbad, CA) viability assay at 1-, 2-, and 3-day intervals.
Toxicity of oligomeric α-synuclein and internalization of α-synuclein
We first investigated the effects of oligomeric α-synuclein on dopaminergic neurons. As shown in Fig. 1A, oligomeric α-synuclein decreased remaining viable neurons significantly over 3 days after treatment. More importantly, the majority of MES cells, though still viable as determined by trypan blue exclusion assay, displayed apparent morphological changes, including shrinkage of cell bodies and dendritic beading after a few hours of treatment. Representative MES cells exposed to oligomeric
Acknowledgments
The study is partially supported by NIH Grants to J.Z. (R01AG025327 and R01ES012703).
References (27)
- et al.
p25alpha Stimulates alpha-synuclein aggregation and is co-localized with aggregated alpha-synuclein in alpha-synucleinopathies
J. Biol. Chem.
(2005) - et al.
Analysis of alpha-synuclein-associated proteins by quantitative proteomics
J. Biol. Chem.
(2004) - et al.
Protein–protein interactions of alpha-synuclein in brain homogenates and transfected cells
Brain Res. Mol. Brain Res.
(2001) - et al.
Tubulin seeds alpha-synuclein fibril formation
J. Biol. Chem.
(2002) - et al.
Proteomic Identification of a stress protein, mortalin/mthsp70/GRP75: relevance to Parkinson’s disease
Mol. Cell Proteomics
(2006) - et al.
Mechanisms of 4-hydroxynonenal-induced neuronal microtubule dysfunction
Brain Res.
(2005) - et al.
FuGENE 6 transfection reagent: the gentle power
Methods
(2004) Effect of ATP on the kinetics of microtubule assembly
J. Biol. Chem.
(1982)DJ-1 up-regulates glutathione synthesis during oxidative stress and inhibits A53T alpha-synuclein toxicity
J. Biol. Chem.
(2005)- et al.
Roles of Drosophila DJ-1 in survival of dopaminergic neurons and oxidative stress
Curr. Biol.
(2005)