Flow cytometric measurement of the cellular propagation of TDP-43 aggregation
Taylor & Francis
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Zeineddine, R., Whiten, D., Farrawell, N., McAlary, L., Hanspal, M., Kumita, J., Wilson, M., & et al. (2017). Flow cytometric measurement of the cellular propagation of TDP-43 aggregation. Prion, 11 (3), 195-204. https://doi.org/10.1080/19336896.2017.1314426
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.
ALS, FloIT, Flow cytometry, Prion, propagation, protein aggregation, TDP-43
This work was supported by the NHMRC under Grant 1084144 and 1095215.
External DOI: https://doi.org/10.1080/19336896.2017.1314426
This record's URL: https://www.repository.cam.ac.uk/handle/1810/265201