The A53T alpha-synuclein mutation increases iron-dependent aggregation and toxicity.
Parkinson's disease (PD) is the most common motor disorder affecting the elderly. PD is characterized by the formation of Lewy bodies and death of dopaminergic neurons. The mechanisms underlying PD are unknown, but the discoveries that mutations in alpha-synuclein can cause familial PD and that alpha-synuclein accumulates in Lewy bodies suggest that alpha-synuclein participates in the pathophysiology of PD. Using human BE-M17 neuroblastoma cells overexpressing wild-type, A53T, or A30P alpha-synuclein, we now show that iron and free radical generators, such as dopamine or hydrogen peroxide, stimulate the production of intracellular aggregates that contain alpha-synuclein and ubiquitin. The aggregates can be identified by immunocytochemistry, electron microscopy, or the histochemical stain thioflavine S. The amount of aggregation occurring in the cells is dependent on the amount of alpha-synuclein expressed and the type of alpha-synuclein expressed, with the amount of alpha-synuclein aggregation following a rank order of A53T > A30P > wild-type > untransfected. In addition to stimulating aggregate formation, alpha-synuclein also appears to induce toxicity. BE-M17 neuroblastoma cells overexpressing alpha-synuclein show up to a fourfold increase in vulnerability to toxicity induced by iron. The vulnerability follows the same rank order as for aggregation. These data raise the possibility that alpha-synuclein acts in concert with iron and dopamine to induce formation of Lewy body pathology in PD and cell death in PD.
Departments of Pharmacology, Loyola University Medical Center, Maywood, Illinois 60153, USA.
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N
Ostrerova-Golts
L
L
Petrucelli
J
J
Hardy
J M
JM
Lee
M
M
Farer
B
B
Wolozin
D052160:Benzothiazoles; D002470:Cell Survival / Q000502:physiology; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism / Q000648:ultrastructure; D007501:Iron / Q000633:toxicity*; D016631:Lewy Bodies / Q000378:metabolism*; D009154:Mutation / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009447:Neuroblastoma; D009474:Neurons / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D018384:Oxidative Stress / Q000502:physiology; D010300:Parkinson Disease / Q000209:etiology / Q000503:physiopathology; D051843:Synucleins; D013844:Thiazoles; D014407:Tumor Cells, Cultured; D014452:Ubiquitins / Q000378:metabolism; D051844:alpha-Synuclein
D052160:Benzothiazoles; D005609:Free Radicals; D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; D051843:Synucleins; D013844:Thiazoles; D014452:Ubiquitins; D051844:alpha-Synuclein; C009462:thioflavin T; D007501:Iron