Interplay of RhoA and mechanical forces in collective cell migration driven by leader cells

The leading front of a collectively migrating epithelium often destabilizes into multicellular migration fingers where a cell initially similar to the others becomes a leader cell while its neighbours do not alter. The determinants of these leader cells include mechanical and biochemical cues, often under the control of small GTPases. However, an accurate dynamic cartography of both mechanical and biochemical activities remains to be established. Here, by mapping the mechanical traction forces exerted on the surface by MDCK migration fingers, we show that these structures are mechanical global entities with the leader cells exerting a large traction force. Moreover, the spatial distribution of RhoA differential activity at the basal plane strikingly mirrors this force cartography. We propose that RhoA controls the development of these fingers through mechanical cues: the leader cell drags the structure and the peripheral pluricellular acto-myosin cable prevents the initiation of new leader cells.

[1]  P. Chavrier,et al.  Collective migration of an epithelial monolayer in response to a model wound , 2007, Proceedings of the National Academy of Sciences.

[2]  M. Matsuda,et al.  Activity of Rho-family GTPases during cell division as visualized with FRET-based probes , 2003, The Journal of cell biology.

[3]  Michael P. Sheetz,et al.  Keratocytes Pull with Similar Forces on Their Dorsal and Ventral Surfaces , 1999, The Journal of cell biology.

[4]  K. Hahn,et al.  RhoA/ROCK-mediated switching between Cdc42- and Rac1-dependent protrusion in MTLn3 carcinoma cells. , 2008, Experimental cell research.

[5]  Bruno Moulia,et al.  In vivo modulation of morphogenetic movements in Drosophila embryos with femtosecond laser pulses. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Eric Mazur,et al.  Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics. , 2006, Biophysical journal.

[7]  E. Sahai,et al.  Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells , 2007, Nature Cell Biology.

[8]  M. Dembo,et al.  Stresses at the cell-to-substrate interface during locomotion of fibroblasts. , 1999, Biophysical journal.

[9]  Gaudenz Danuser,et al.  Coordination of Rho GTPase activities during cell protrusion , 2009, Nature.

[10]  M. Poujade,et al.  Velocity fields in a collectively migrating epithelium. , 2010, Biophysical journal.

[11]  Gaudenz Danuser,et al.  Protrusion and actin assembly are coupled to the organization of lamellar contractile structures. , 2010, Experimental cell research.

[12]  Yi I. Wu,et al.  Light-mediated activation reveals a key role for Rac in collective guidance of cell movement in vivo , 2010, Nature Cell Biology.

[13]  A. Hall,et al.  Myosin-IXA Regulates Collective Epithelial Cell Migration by Targeting RhoGAP Activity to Cell-Cell Junctions , 2012, Current Biology.

[14]  Jean-Jacques Meister,et al.  Force transmission in migrating cells , 2010, The Journal of cell biology.

[15]  Marion Ghibaudo,et al.  Traction forces and rigidity sensing regulate cell functions , 2008 .

[16]  Y. Hegerfeldt,et al.  Collective cell movement in primary melanoma explants: plasticity of cell-cell interaction, beta1-integrin function, and migration strategies. , 2002, Cancer research.

[17]  K. Burridge,et al.  From mechanical force to RhoA activation. , 2012, Biochemistry.

[18]  S. Atkinson,et al.  Rho GTPase signaling regulates tight junction assembly and protects tight junctions during ATP depletion. , 1998, American journal of physiology. Cell physiology.

[19]  Shirley Mark,et al.  Physical model of the dynamic instability in an expanding cell culture. , 2010, Biophysical journal.

[20]  E. Grasland-Mongrain,et al.  Orientation and polarity in collectively migrating cell structures: statics and dynamics. , 2011, Biophysical journal.

[21]  Paul Martin,et al.  Wound healing recapitulates morphogenesis in Drosophila embryos , 2002, Nature Cell Biology.

[22]  J. Fredberg,et al.  Collective cell guidance by cooperative intercellular forces , 2010, Nature materials.

[23]  M. Matsuda,et al.  Activation of Rac and Cdc42 Video Imaged by Fluorescent Resonance Energy Transfer-Based Single-Molecule Probes in the Membrane of Living Cells , 2002, Molecular and Cellular Biology.

[24]  P. Friedl,et al.  Collective cell migration in morphogenesis, regeneration and cancer , 2009, Nature Reviews Molecular Cell Biology.

[25]  K. Hahn,et al.  Spatiotemporal dynamics of RhoA activity in migrating cells , 2006, Nature.

[26]  A. Ridley Life at the Leading Edge , 2011, Cell.

[27]  T. Meyer,et al.  Modular control of endothelial sheet migration. , 2008, Genes & development.

[28]  Peter Friedl,et al.  Determinants of leader cells in collective cell migration. , 2010, Integrative biology : quantitative biosciences from nano to macro.

[29]  M. Sheetz,et al.  Two distinct modes of myosin assembly and dynamics during epithelial wound closure , 2007, The Journal of cell biology.

[30]  S. Atkinson,et al.  Rho-kinase regulates myosin II activation in MDCK cells during recovery after ATP depletion. , 2001, American journal of physiology. Renal physiology.

[31]  J. Klarlund Dual modes of motility at the leading edge of migrating epithelial cell sheets , 2012, Proceedings of the National Academy of Sciences.

[32]  I M Gelfand,et al.  Rho-dependent formation of epithelial “leader” cells during wound healing , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[33]  M. Parsons,et al.  Collective Chemotaxis Requires Contact-Dependent Cell Polarity , 2010, Developmental cell.

[34]  P. Rørth Collective guidance of collective cell migration. , 2007, Trends in cell biology.

[35]  Suliana Manley,et al.  A role for actin arcs in the leading-edge advance of migrating cells , 2011, Nature Cell Biology.

[36]  R. Austin,et al.  Force mapping in epithelial cell migration. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Timothy J Mitchison,et al.  Dissecting Temporal and Spatial Control of Cytokinesis with a Myosin II Inhibitor , 2003, Science.

[38]  E. Fuchs,et al.  Actin cable dynamics and Rho/Rock orchestrate a polarized cytoskeletal architecture in the early steps of assembling a stratified epithelium. , 2002, Developmental cell.

[39]  J. Xu,et al.  Differential RhoA Dynamics in Migratory and Stationary Cells Measured by FRET and Automated Image Analysis , 2008, PloS one.

[40]  William I. Weis,et al.  Deconstructing the Cadherin-Catenin-Actin Complex , 2005, Cell.

[41]  P. Friedl,et al.  Classifying collective cancer cell invasion , 2012, Nature Cell Biology.

[42]  C. McCulloch,et al.  Force activates smooth muscle α-actin promoter activity through the Rho signaling pathway , 2007, Journal of Cell Science.

[43]  J. Thiery,et al.  A scatter factor-like factor is produced by a metastatic variant of a rat bladder carcinoma cell line. , 1994, Journal of cell science.

[44]  Shu Chien,et al.  Cooperative effects of Rho and mechanical stretch on stress fiber organization. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[45]  E. Sahai,et al.  Collective cell migration requires suppression of actomyosin at cell-cell contacts mediated by DDR1 and the cell polarity regulators Par3 and Par6 , 2010, Nature Cell Biology.

[46]  Pascal Silberzan,et al.  Automated velocity mapping of migrating cell populations (AVeMap) , 2012, Nature Methods.

[47]  Rizwan U. Farooqui,et al.  Multiple rows of cells behind an epithelial wound edge extend cryptic lamellipodia to collectively drive cell-sheet movement , 2005, Journal of Cell Science.

[48]  G. Edwards,et al.  Forces for Morphogenesis Investigated with Laser Microsurgery and Quantitative Modeling , 2003, Science.

[49]  Srivatsan Raghavan,et al.  Cell polarity triggered by cell-cell adhesion via E-cadherin , 2009, Journal of Cell Science.

[50]  Kazuhiro Aoki,et al.  Visualization of small GTPase activity with fluorescence resonance energy transfer-based biosensors , 2009, Nature Protocols.

[51]  M. Matsuda,et al.  Localized RhoA activation as a requirement for the induction of membrane ruffling. , 2005, Molecular biology of the cell.

[52]  David A. Weitz,et al.  Physical forces during collective cell migration , 2009 .

[53]  K. Hahn,et al.  Imaging and photobleach correction of Mero-CBD, sensor of endogenous Cdc42 activation. , 2006, Methods in enzymology.

[54]  Pascal Silberzan,et al.  Is the mechanical activity of epithelial cells controlled by deformations or forces? , 2005, Biophysical journal.