Open AccessSingle cell imaging and quantification of TDP-43 and α-synuclein intercellular propagation
Author(s) -
Sivan Peled,
Dorin Sade,
Yaron Bram,
Ziv Porat,
Topaz Kreiser,
Michael Mimouni,
Alexandra Lichtenstein,
Daniel Segal,
Ehud Gazit
Publication year - 2017
Publication title -
scientific reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.24
H-Index - 213
ISSN - 2045-2322
DOI - 10.1038/s41598-017-00657-z
Subject(s) - flow cytometry , cell , confocal microscopy , intracellular , microbiology and biotechnology , microvesicles , protein subcellular localization prediction , confocal , biology , biophysics , chemistry , computational biology , biochemistry , gene , microrna , geometry , mathematics
The intercellular spreading of protein assemblies is a major factor in the progression of neurodegenerative disorders. The quantitative study and visualization of cell-to-cell propagation using tagged-proteins is challenging due to the steric effect of relatively large fluorescence tags and the risk of ‘false positive’ identification when analyzing these rare transmission events. Here, we established a cell culture model to characterize the cell-to-cell transmission of TAR DNA-binding protein and α-synuclein, involved in amyotrophic lateral sclerosis and Parkinson’s disease, respectively, using the small nine amino acid influenza hemagglutinin tag. The novel use of single cell resolution imaging flow cytometry allowed the visualization and quantification of all individual transmission events. Cell-level analysis of these events indicated that the degree of transfer is lower than previously reported based on conventional flow cytometry. Furthermore, our analysis can exclude ‘false positive’ events of cellular overlap and extracellular debris attachment. The results were corroborated by high-resolution confocal microscopy mapping of protein localization.