Abstract:
: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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Peled S,Sade D,Bram Y,Porat Z,Kreiser T,Mimouni M,Lichtenstein A,Segal D,Gazit Edoi
10.1038/s41598-017-00657-zsubject
Has Abstractpub_date
2017-03-28 00:00:00pages
544issue
1issn
2045-2322pii
10.1038/s41598-017-00657-zjournal_volume
7pub_type
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