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Prion. 2017 May 4;11(3):195-204. doi: 10.1080/19336896.2017.1314426. Epub 2017 May 9.

Flow cytometric measurement of the cellular propagation of TDP-43 aggregation.

Author information

1
a Illawarra Health and Medical Research Institute , Wollongong , NSW , Australia.
2
b School of Biological Sciences, Science Medicine and Health Faculty , University of Wollongong, Wollongong , NSW , Australia.
3
c Department of Chemistry , University of Cambridge , Cambridge , UK.
4
d Centre for Misfolding Diseases , Cambridge , UK.

Abstract

Amyotrophic lateral sclerosis is a devastating neuromuscular degenerative disease characterized by a focal onset of motor neuron loss, followed by contiguous outward spreading of pathology including TAR DNA-binding protein of 43 kDa (TDP-43) aggregates. Previous work suggests that TDP-43 can move between cells. Here we used a novel flow cytometry technique (FloIT) to analyze TDP-43 inclusions and propagation. When cells were transfected to express either mutant G294A TDP-43 fused to GFP or wild type TDP-43fused to tomato red and then co-cultured, flow cytometry detected intact cells containing both fusion proteins and using FloIT detected an increase in the numbers of inclusions in lysates from cells expressing wild type TDP-43-tomato. Furthermore, in this same model, FloIT analyses detected inclusions containing both fusion proteins. These results imply the transfer of TDP-43 fusion proteins between cells and that this process can increase aggregation of wild-type TDP-43 by a mechanism involving co-aggregation with G294A TDP-43.

KEYWORDS:

ALS; FloIT; Flow cytometry; Prion; Propagation; Protein Aggregation; TDP-43

PMID:
28486039
PMCID:
PMC5480386
DOI:
10.1080/19336896.2017.1314426
[Indexed for MEDLINE]
Free PMC Article

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