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Measuring Cell-Edge Protrusion Dynamics during Spreading using Live-Cell Microscopy

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Subcellular and Dynamic Coordination between Src Activity and Cell Protrusion in Microenvironment.

Yue Zhuo¹, Tongcheng Qian¹, Yiqian Wu²

¹Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.

Scientific Reports

|August 12, 2015

Summary

This summary is machine-generated.

Endothelial cell migration, crucial for wound healing and angiogenesis, involves Src kinase. This study reveals Src deactivation coordinates with cell edge protrusion during migration initiation, particularly in polarized cells.

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Area of Science:

Cell Biology
Biophysics
Biochemistry

Background:

Endothelial cell migration is vital for wound healing and angiogenesis.
Src kinase activity is implicated in the protrusions of migrating endothelial cells.
The precise spatiotemporal regulation of Src activity during cell edge protrusion is not fully understood.

Purpose of the Study:

To investigate the spatiotemporal coordination between Src kinase activity and endothelial cell edge protrusion at the initiation of cell migration.
To elucidate the role of physical constraints in regulating these coordinated events.
To develop a new model for Src kinase regulation during endothelial migration.

Main Methods:

Integration of microfabrication techniques to control physical constraints.
Utilization of fluorescence resonance energy transfer (FRET)-based biosensors to monitor Src kinase activity in real-time.
Application of automated computational image analysis for quantitative assessment of cell migration dynamics.

Main Results:

The release of physical restrictive micropatterns led to a significant decrease in Src kinase activity at the endothelial cell protrusive edge.
Computational cross-correlation analysis demonstrated that Src activity decrease preceded and coordinated with cell edge protrusion in polarized cells.
This coordination was not observed in non-polarized cells, highlighting the role of cell polarization.

Conclusions:

Src kinase activity is spatiotemporally controlled and coordinated with cell polarization and protrusion during endothelial cell migration following the release of physical constraints.
This finding offers insights into endothelial cell behavior in contexts like angiogenesis within stiff tumor microenvironments.
The study presents a novel model where Src deactivation is coupled with membrane protrusion, advancing the understanding of endothelial migration regulation.