Amyotrophic lateral sclerosis (ALS) is one of the most common degenerative disease of the motor neuron system. The TDP-43 pathology seems to be a dominant type of pathology across sporadic ALS types. Our TDP-43 Stress Granules Assay cell line allows the quantification of pathological TDP43 globs into the nucleus and cytosol. This model also permits to monitor the TDP-43 protein distribution in living cells; studying the protein localization pattern in the space and time, and quantifying the fluorescence aggregation inside the cells.
To perform the assay, we induce fluorescent TDP-43 expression adding IPTG to the cell culture medium. After TDP-43 expression induction and during the assay, we incubate the cells with test compounds and Sodium Arsenite to induce the TDP-43 aggregation. Finally, we quantify TDP43-tGFP nuclear globs using the BD Pathway HCS Reader and Attovision Compartimentalization Software.
- TDP-43 Stress Granules Assay cell line allows the quantification of pathological TDP43 globs.
- This model permits to evaluate the TDP-43 protein distribution in living cells studying the protein localization pattern in the space and time.
- This model provides a strategy to evaluate drugs without cell permeability.
- This cellular model have been adapted to HCS analyses based on image algorithms to test cytosolic and nuclear globs generation process.