Accelerated α-synuclein aggregation after differentiation of SH-SY5Y neuroblastoma cells

• Published in: Brain research Vol. 1013; no. 1; pp. 51 - 59
• Main Authors: Hasegawa, Takafumi, Matsuzaki, Michiko, Takeda, Atsushi, Kikuchi, Akio, Akita, Hirotoshi, Perry, George, Smith, Mark A, Itoyama, Yasuto
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 02.07.2004; Amsterdam Elsevier; New York, NY
• Subjects: Aggregation; alpha-Synuclein; Biological and medical sciences; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Line, Tumor; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; Humans; Iron; Lewy body; Medical sciences; Microtubules - metabolism; Microtubules - pathology; Nerve Tissue Proteins - biosynthesis; Nerve Tissue Proteins - metabolism; Neuroblastoma; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neurology; Parkinson's disease; Synucleins; α-Synuclein; Disorders of the nervous system; α-Synuclein; Parkinson's disease; UCH-L1; AGPC; Iron; Neuroblastoma; WB; Aggregation; DMEM; TUNEL; FBS; RT-PCR; WT; BDNF; MT; RA; mAb; ECL; pAb; PD; α-syn; CAT; LBs; GAPDH; Lewy body; Nervous system diseases; Pathogenesis; Parkinson disease; Cerebral disorder; Cell line; Neuron; Central nervous system disease; Degenerative disease; Differentiation; Extrapyramidal syndrome
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2004.04.018

α-Synuclein (α-syn) is a major component of inclusion bodies in Parkinson's disease (PD) and other synucleinopathies. To clarify the possible roles of α-syn in the molecular pathogenesis of neurodegenerative diseases, we have established a novel cellular model based on the differentiation of SH-SY5Y cells that overexpress α-syn. In the presence of ferrous iron, differentiation of the cells led to the formation of large perinuclear inclusion bodies, which developed from scattered small aggregates seen in undifferentiated cells. The iron-induced α-syn-positive inclusions co-localized largely with ubiquitin, and some of them were positive for nitrotyrosine, lipid, γ-tubulin and dynein. Notably, treatment with nocodazole, a microtubule depolymerizing agent, interrupted the aggregate formation but led to a concomitant increase of apoptotic cells. Therefore, it appears that an intracellular retrograde transport system via microtubules plays a crucial role in the aggregate formation and also that the aggregates may represent a cytoprotective response against noxious stimuli. This cellular model will enable better understanding of the molecular pathomechanisms of synucleinopathy.