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Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis
Published on: July 3, 2015
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Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis
Edmund Ong1, Catherine Cahill2
1Department of Anesthesiology and Perioperative Care, University of California Irvine; Department of Biomedical and Molecular Sciences, Queen's University; edmund.ong@uci.edu.
Journal of Visualized Experiments : Jove
|July 14, 2015
Summary
This study presents a new method to track receptor trafficking in cells using immunolabelling and confocal microscopy. This technique quantitatively assesses how drug treatments alter receptor movement within cells.
Area of Science:
- Cell biology
- Molecular biology
- Biochemistry
Background:
- Intracellular receptor trafficking is crucial for cell signaling and responsiveness.
- Receptor trafficking is dynamically regulated by both internal and external cellular conditions.
- Understanding receptor trafficking is key to developing targeted therapies.
Purpose of the Study:
- To develop and validate a robust protocol for quantitatively assessing induced intracellular receptor trafficking.
- To provide a flexible method applicable to cultured cells and tissue slices.
- To enable the analysis of trafficking changes following various interventions, including drug treatments.
Main Methods:
- Utilizes double-immunolabelling for receptors and intracellular compartment markers.
- Employs sequential confocal microscopy for capturing cellular images.
- Applies computerized colocalizational analysis to determine quantitative colocalization scores.
Main Results:
- The protocol yields normalized colocalization scores for statistical analysis.
- Allows for the quantitative assessment of receptor trafficking changes after drug treatment.
- Provides a powerful and flexible technique for induced receptor trafficking studies.
Conclusions:
- This method offers a powerful and flexible approach to quantitatively assess induced receptor trafficking.
- The protocol is optimized for cultured cells but adaptable to tissue slices.
- Enables detailed analysis of how cellular interventions impact receptor localization and signaling.
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