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Quantifying autophagy using novel LC3B and p62 TR-FRET assays
Alberto Bresciani1, Maria Carolina Spiezia1, Roberto Boggio2
1IRBM Science Park, Pomezia, Rome, Italy.
Plos One
|March 20, 2018
Summary
Novel assays measuring autophagy, a key cellular process, were developed. These tools aid in discovering new treatments for neurodegenerative diseases by identifying autophagy upregulators.
Area of Science:
- Cell Biology
- Biochemistry
- Neuroscience
Background:
- Autophagy is a cellular process vital for energy generation and clearing misfolded proteins.
- Upregulating autophagy is a potential therapeutic strategy for neurodegenerative diseases.
Purpose of the Study:
- To develop novel assays for detecting changes in autophagy.
- To facilitate high-throughput screening for autophagy upregulators.
Main Methods:
- Development of time-resolved fluorescence resonance energy transfer (TR-FRET) assays using LC3B-II and p62.
- Assays detect autophagy changes without exogenous labels in cells and tissues.
Main Results:
- Novel TR-FRET assays successfully measure autophagy markers LC3B-II and p62.
- Demonstrated varying cellular responses to pharmacological autophagy interventions.
- Highlighted the necessity of multiple readouts for accurate autophagy assessment.
Conclusions:
- The developed TR-FRET assays are effective tools for autophagy research.
- These assays can accelerate the discovery of novel therapeutic agents for neurodegenerative conditions.
- Comprehensive assessment using multiple readouts is crucial for understanding autophagy modulation.
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