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Sensitive Detection of Proteopathic Seeding Activity with FRET Flow Cytometry
Published on: December 8, 2015
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A seeding based cellular assay of tauopathy.
Yin Xu1, Heidi Martini-Stoica1,2,3, Hui Zheng4,5,6
1Huffington Center on Aging, Baylor College of Medicine, Houston, TX, USA.
Molecular Neurodegeneration
|April 27, 2016
Summary
Researchers developed a novel cellular model for tauopathy by mimicking prion-like mechanisms. This model effectively replicates neurofibrillary tangles in vitro, aiding the study of tau protein aggregation and the identification of potential therapeutic modulators.
Area of Science:
- Neuroscience
- Biochemistry
Background:
- Tauopathy is a neurodegenerative disease defined by neurofibrillary tangles of hyperphosphorylated tau.
- Existing in vitro cellular models do not adequately replicate neurofibrillary tangles.
- Neurofibrillary tangle formation is hypothesized to occur via a prion replication mechanism.
Purpose of the Study:
- To establish a seeding-based cellular model that mimics neurofibrillary tangle pathology in vitro.
- To investigate the role of autophagy in the formation of insoluble tau.
- To develop a high-content screening platform for identifying tau modulators.
Main Methods:
- Utilized brain lysate from rTg4510 mice and synthetic preformed tau fibrils (PFFs) as seeds.
- Converted soluble tau to insoluble tau within the cellular model.
- Manipulated autophagy levels using genetic and pharmacological approaches.
- Quantified insoluble tau using thioflavin-S staining.
Main Results:
- The cellular model successfully generated insoluble tau with characteristics of neurofibrillary tangles.
- Inhibition of autophagy increased insoluble tau, while enhancement of autophagy reduced it.
- The model is amenable to high-content screening for tau modulators.
Conclusions:
- The developed seeding-based cellular model accurately reproduces in vitro neurofibrillary tangle pathology.
- This model serves as a valuable tool for tauopathy research.
- The platform facilitates the identification of therapeutic agents targeting tau pathology.
