Title: P3‐057: A novel approach for monitoring tau aggregation in cell models of tauopathy by fluorescence‐activated cell sorting (FACS)
Abstract: The inhibition or reversal of tau aggregation is of potential therapeutic importance for Alzheimer's Disease and other tauopathies. We have previously generated a cell model of tauopathy based on the expression of an amyloidogenic tau construct in neuroblastoma N2a cells (Khlistunova et al., JBC 2006). Thus far, the quantitation of aggregation was based on fluorescence microscopy of cells stained with the dye thioflavin S. We developed a FACS-based assay of tau aggregation in cells for rapid evaluation of tau aggregation inhibitors and for separating cells in different stages of aggregation for biochemical analysis. Using FACS we have precisely counted N2a cells containing ThS-positive tau aggregates. Moreover FACS allowed us the measurement of the effect of inhibitory compounds on tau aggregation within N2a cells and the establishment of dose response relationships. The growing number of N2a cells containing aggregated tau repeat domain correlated very well with decreasing viability measured by MTT and increasing toxicity evaluated by LDH assays. Moreover, we have applied FACS for the preparative separation of cells containing tau aggregates from non-fluorescent cells without aggregates and will present a biochemical analysis of the protein modifications in both cases. The improvements in handling and separating of large cell numbers in a cell model of tauopathy has allowed us to speed up the design of cell-permeable compounds with low toxicity for the development of novel tau aggregation inhibitors.
Publication Year: 2011
Publication Date: 2011-07-01
Language: en
Type: article
Indexed In: ['crossref']
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