Subcellular and Dynamic Coordination between Src Activity and Cell Protrusion in Microenvironment.
Migration of endothelial cells is essential for wound healing and angiogenesis. Src kinase activity plays important roles at the protrusions of migrating endothelial cells. However, the spatiotemporal coordination between Src kinase activity and the protrusion of cell edge remains unclear. Therefore, we investigate these coordinated molecular events at the initiation of cell migration, by integrating microfabrication, fluorescence resonance energy transfer (FRET)-based biosensors, and automated computational image analysis. We demonstrate that the physical release of restrictive micropattern triggered a significant decrease of Src activity at the protrusive edge of endothelial cells. Computational cross-correlation analysis reveals that the decrease of Src activity occurred earlier in time, and was well-coordinated with the protrusion of cell edge in polarized cells, but not in non-polarized cells. These results suggest that the spatiotemporal control of Src kinase activity is well-coordinated with cell polarization and protrusion in endothelial cells upon the release of physical constraint, as that experienced by endothelial cells sprouting from stiff tumor micro-environment during angiogenesis. Therefore, our integrative approach enabled the discovery of a new model where Src is de-activated in coordination with membrane protrusion, providing important insights into the regulation of endothelial migration and angiogenesis.
Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Yue
Y
Zhuo
Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Tongcheng
T
Qian
Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093-0435.
Yiqian
Y
Wu
1] Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 [2] Center for Neuro-Medicine, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, South Korea.
Jihye
J
Seong
Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093-0435.
Ya
Y
Gong
Suzhou Institute of Nano-Tech and Nano-Bionics, Suzhou, China.
Hongwei
H
Ma
1] Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 [2] Neuroscience Program, Beckman Institute for Advanced Science and Technology, Department of Molecular and Integrative Physiology and University of Illinois at Urbana-Champaign, Urbana, IL 61801 [3] Center of Biophysics and Computational Biology, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 [4] Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093-0435.
Yingxiao
Y
Wang
1] Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 [2] Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093-0435.
Shaoying
S
Lu
D002465:Cell Movement / Q000235:genetics; D016764:Cell Polarity / Q000235:genetics; D022081:Cell Surface Extensions / Q000235:genetics; D060833:Cellular Microenvironment / Q000235:genetics*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D018919:Neovascularization, Physiologic / Q000235:genetics; D014945:Wound Healing / Q000235:genetics*; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
D019061:src-Family Kinases