Elucidation of characters and neurotoxic mechanisms of alpha-synuclein oligomers and its application to molecular targeted therapy

点击翻译按钮获取中文摘要

We established the preparing method of alpha-synuclein (A-syn) oligomers and protofibrils by incubating high-concentration solution, 3 mg/mL, of recombinant A-syn with stirring for 7 days at 37 C. Although recombinant A-syn molecules and natural A-syn molecules derived from human CSF were eluted from gel-filtration chromatography at around fractions of 60-70 kDa, those molecules were not dissociated with 8M urea, and accordingly have been demonstrated to exist as monomers in aqueous solution. In cultured cells, intracellular aggregates were formed by adding mixture of recombinant A-syn oligomers and protofibrils into the culture medium, and their degradation was dependent on the autophagy pathway.