Direct Observation of α-Synuclein Amyloid Aggregates in Endocytic Vesicles of Neuroblastoma Cells

Mihaela M Apetri, Rolf Harkes, Vinod Subramaniam, Gerard W Canters, Thomas Schmidt, Thijs J Aartsma

    Research output: Contribution to JournalArticleAcademicpeer-review

    Abstract

    Aggregation of α-synuclein has been linked to both familial and sporadic Parkinson's disease. Recent studies suggest that α-synuclein aggregates may spread from cell to cell and raise questions about the propagation of neurodegeneration. While continuous progress has been made characterizing α-synuclein aggregates in vitro, there is a lack of information regarding the structure of these species inside the cells. Here, we use confocal fluorescence microscopy in combination with direct stochastic optical reconstruction microscopy, dSTORM, to investigate α-synuclein uptake when added exogenously to SH-SY5Y neuroblastoma cells, and to probe in situ morphological features of α-synuclein aggregates with near nanometer resolution. We demonstrate that using dSTORM, it is possible to follow noninvasively the uptake of extracellularly added α-synuclein aggregates by the cells. Once the aggregates are internalized, they move through the endosomal pathway and accumulate in lysosomes to be degraded. Our dSTORM data show that α-synuclein aggregates remain assembled after internalization and they are shortened as they move through the endosomal pathway. No further aggregation was observed inside the lysosomes as speculated in the literature, nor in the cytoplasm of the cells. Our study thus highlights the super-resolution capability of dSTORM to follow directly the endocytotic uptake of extracellularly added amyloid aggregates and to probe the morphology of in situ protein aggregates even when they accumulate in small vesicular compartments.

    Original languageEnglish
    Pages (from-to)e0153020
    JournalPLoS ONE
    Volume11
    Issue number4
    DOIs
    Publication statusPublished - 2016

    Keywords

    • Amyloid
    • Cell Line, Tumor
    • Endocytosis
    • Humans
    • Microscopy, Atomic Force
    • Neuroblastoma
    • alpha-Synuclein
    • Journal Article

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