title: Vinculin is required for cell polarization, migration, and extracellular matrix remodeling in 3D collagen.
abstract: Vinculin is filamentous (F)-actin-binding protein enriched in integrin-based adhesions to the extracellular matrix (ECM). Whereas studies in 2-dimensional (2D) tissue culture models have suggested that vinculin negatively regulates cell migration by promoting cytoskeleton-ECM coupling to strengthen and stabilize adhesions, its role in regulating cell migration in more physiologic, 3-dimensional (3D) environments is unclear. To address the role of vinculin in 3D cell migration, we analyzed the morphodynamics, migration, and ECM remodeling of primary murine embryonic fibroblasts (MEFs) with cre/loxP-mediated vinculin gene disruption in 3D collagen I cultures. We found that vinculin promoted 3D cell migration by increasing directional persistence. Vinculin was necessary for persistent cell protrusion, cell elongation, and stable cell orientation in 3D collagen, but was dispensable for lamellipodia formation, suggesting that vinculin-mediated cell adhesion to the ECM is needed to convert actin-based cell protrusion into persistent cell shape change and migration. Consistent with this finding, vinculin was necessary for efficient traction force generation in 3D collagen without affecting myosin II activity and promoted 3D collagen fiber alignment and macroscopical gel contraction. Our results suggest that vinculin promotes directionally persistent cell migration and tension-dependent ECM remodeling in complex 3D environments by increasing cell-ECM adhesion and traction force generation.
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA ithievessen@biomed.uni-erlangen.de.
authors.forename: Ingo
authors.initials: I
authors.lastname: Thievessen
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Nikta
authors.initials: N
authors.lastname: Fakhri
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Julian
authors.initials: J
authors.lastname: Steinwachs
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Viola
authors.initials: V
authors.lastname: Kraus
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: R Scott
authors.initials: RS
authors.lastname: McIsaac
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Liang
authors.initials: L
authors.lastname: Gao
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Bi-Chang
authors.initials: BC
authors.lastname: Chen
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Baird
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Betzig
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Oldenbourg
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Clare M
authors.initials: CM
authors.lastname: Waterman
authors.affiliation: *Laboratory of Cell and Tissue Morphodynamics, Cell Biology and Physiology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA; Biophysics Group, Department of Physics, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany; Physiology Course and Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA; Third Physics Institute-Biophysics, Georg-August-University, Göttingen, Germany; Physics of Living Systems, Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA; California Life Company, South San Francisco, California, USA; **Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA; Department of Chemistry, Stony Brook University, Stony Brook, New York, USA; Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan; Department of Biological Science, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Ben
authors.initials: B
authors.lastname: Fabry
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology*; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism; D011554:Pseudopodia / Q000235:genetics / Q000378:metabolism; D016596:Vinculin / Q000235:genetics / Q000378:metabolism*
chemical_list: D016596:Vinculin; D003094:Collagen; D024462:Myosin Type II
keywords: 3D cell migration; cell morphodynamics; integrin; traction force generation