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Updated: May 25, 2026
Imaging G-protein Coupled Receptor (GPCR)-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
Published on: September 20, 2011
Imaging the impact of chemically inducible proteins on cellular dynamics in vivo
Hon S Leong1, Michael M Lizardo, Amber Ablack
1Translational Prostate Cancer Research Group, London Regional Cancer Program, London, Ontario, Canada.
Abstract:
The analysis of dynamic events in the tumor microenvironment during cancer progression is limited by the complexity of current in vivo imaging models. This is coupled with an inability to rapidly modulate and visualize protein activity in real time and to understand the consequence of these perturbations in vivo. We developed an intravital imaging approach that allows the rapid induction and subsequent depletion of target protein levels within human cancer xenografts while assessing the impact on cell behavior and morphology in real time. A conditionally stabilized fluorescent E-cadherin chimera was expressed in metastatic breast cancer cells, and the impact of E-cadherin induction and depletion was visualized using real-time confocal microscopy in a xenograft avian embryo model. We demonstrate the assessment of protein localization, cell morphology and migration in cells undergoing epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors. This technique allows for precise control over protein activity in vivo while permitting the temporal analysis of dynamic biophysical parameters.
Insights
This study introduces a novel intravital imaging technique to track dynamic changes in the tumor microenvironment. The method allows real-time visualization and modulation of protein activity, offering new insights into cancer progression.
Area of Science:
- Oncology
- Cell Biology
- Biophysics
Background:
- Analyzing dynamic events in the tumor microenvironment during cancer progression is challenging due to complex in vivo imaging models.
- Current methods struggle with rapid modulation and real-time visualization of protein activity and its in vivo consequences.
Purpose of the Study:
- To develop an intravital imaging approach for real-time modulation and visualization of protein activity in human cancer xenografts.
- To assess the impact of protein perturbations on cell behavior, morphology, and migration in vivo.
Main Methods:
- Developed a method for rapid induction and depletion of target protein levels within human cancer xenografts.
- Utilized a conditionally stabilized fluorescent E-cadherin chimera in metastatic breast cancer cells.
- Employed real-time confocal microscopy in a xenograft avian embryo model for visualization.
Main Results:
- Demonstrated visualization of protein localization, cell morphology, and migration during epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors.
- Showcased precise control over protein activity in vivo.
- Enabled temporal analysis of dynamic biophysical parameters.
Conclusions:
- The developed intravital imaging technique overcomes limitations in studying dynamic tumor microenvironment events.
- This approach allows for precise temporal control and analysis of protein activity and its effects on cancer cell behavior in vivo.
