Familial Parkinson's Disease Mutant E46K α-Synuclein Localizes to Membranous Structures, Forms Aggregates, and Induces Toxicity in Yeast Models
Figure 3
E46K α-synuclein exhibits strain-specific aggregation and endomembrane localization in fission yeast. (a and b) Live cell GFP microscopy of TCP1 cells (a) and SP3 cells (b) expressing WT, E46K, A30P, or A53T α-synuclein, and GFP alone, at 24 and 48 hours of expression (left). Quantification: ~750 cells of each type were scored for these localization patterns: cytoplasmically diffuse, aggregate, endomembrane, endomembrane/diffuse, endomembrane/aggregate (right). Phenotypes were plotted as a percent of total cells that fluoresced . In TCP1, E46K α-synuclein mostly aggregated in TCP1, like WT and A53T. In SP3, E46K α-synuclein became prominenty localized to endomembrane systems, more so than WT and A53T. A30P was cytoplasmically diffuse like GFP in both strains. (c) E46K aggregates mostly varied from 1–5 per cell, but as many as 10 aggregates per cell were observed. (d) DAPI staining indicated that E46K localization in both strains (TCP1: top; SP3: bottom) was outside the nucleus. (e) E46K cells often exhibited prevacuolar localization and vesicular accumulation.
