title: Spatio-temporal co-ordination of RhoA, Rac1 and Cdc42 activation during prototypical edge protrusion and retraction dynamics.
abstract: 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.
authors.affiliation: Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
authors.forename: Katrin
authors.initials: K
authors.lastname: Martin
authors.affiliation: Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Reimann
authors.affiliation: Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
authors.forename: Rafael D
authors.initials: RD
authors.lastname: Fritz
authors.affiliation: School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, 151-742, Republic of Korea.|Institute of Advanced Machinery and Design, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Hyunryul
authors.initials: H
authors.lastname: Ryu
authors.affiliation: School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, 151-742, Republic of Korea.|Institute of Advanced Machinery and Design, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Noo Li
authors.initials: NL
authors.lastname: Jeon
authors.affiliation: Dept. of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland.
authors.forename: Olivier
authors.initials: O
authors.lastname: Pertz
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects; D000199:Actins / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D000818:Animals; D015374:Biosensing Techniques; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D008856:Microscopy, Fluorescence; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D000577:Amides; D005456:Fluorescent Dyes; D010982:Platelet-Derived Growth Factor; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein