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Spatio-temporal co-ordination of RhoA, Rac1 and Cdc42 activation during prototypical edge protrusion and retraction dynamics.

  • Martin K Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
  • Reimann A Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
  • Fritz RD Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
  • Ryu H School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, 151-742, Republic of Korea.
  • Jeon NL School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, 151-742, Republic of Korea.
  • Pertz O Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
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  • 2016-02-26
Published in:
  • Scientific reports. - 2016
Actin Cytoskeleton
Actins
Amides
Animals
Biosensing Techniques
Cell Adhesion
Cell Line
Cell Movement
Fibroblasts
Fluorescence Resonance Energy Transfer
Fluorescent Dyes
Microfluidic Analytical Techniques
Microscopy, Fluorescence
Platelet-Derived Growth Factor
Pyridines
Rats
cdc42 GTP-Binding Protein
rac1 GTP-Binding Protein
rho-Associated Kinases
rhoA GTP-Binding Protein
English The three canonical Rho GTPases RhoA, Rac1 and Cdc42 co-ordinate cytoskeletal dynamics. Recent studies indicate that all three Rho GTPases are activated at the leading edge of motile fibroblasts, where their activity fluctuates at subminute time and micrometer length scales. Here, we use a microfluidic chip to acutely manipulate fibroblast edge dynamics by applying pulses of platelet-derived growth factor (PDGF) or the Rho kinase inhibitor Y-27632 (which lowers contractility). This induces acute and robust membrane protrusion and retraction events, that exhibit stereotyped cytoskeletal dynamics, allowing us to fairly compare specific morphodynamic states across experiments. Using a novel Cdc42, as well as previously described, second generation RhoA and Rac1 biosensors, we observe distinct spatio-temporal signaling programs that involve all three Rho GTPases, during protrusion/retraction edge dynamics. Our results suggest that Rac1, Cdc42 and RhoA regulate different cytoskeletal and adhesion processes to fine tune the highly plastic edge protrusion/retraction dynamics that power cell motility.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/124540
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