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Fibroblast polarization is a matrix-rigidity-dependent process controlled by focal adhesion mechanosensing

Nature Cell Biology volume 13, pages 1457–1465 (2011)Cite this article

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Abstract

Cell elongation and polarization are basic morphogenetic responses to extracellular matrix adhesion. We demonstrate here that human cultured fibroblasts readily polarize when plated on rigid, but not on compliant, substrates. On rigid surfaces, large and uniformly oriented focal adhesions are formed, whereas cells plated on compliant substrates form numerous small and radially oriented adhesions. Live-cell monitoring showed that focal adhesion alignment precedes the overall elongation of the cell, indicating that focal adhesion orientation may direct cell polarization. siRNA-mediated knockdown of 85 human protein tyrosine kinases (PTKs) induced distinct alterations in the cell polarization response, as well as diverse changes in cell traction force generation and focal adhesion formation. Remarkably, changes in rigidity-dependent traction force development, or focal adhesion mechanosensing, were consistently accompanied by abnormalities in the cell polarization response. We propose that the different stages of cell polarization are regulated by multiple, PTK-dependent molecular checkpoints that jointly control cell contractility and focal-adhesion-mediated mechanosensing.

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Figure 1: Fibroblasts polarize on rigid, but not on compliant, substrates.
Figure 2: Characterization of focal adhesions in cells on stiff and compliant substrates.
Figure 3: Examples of candidate knockdowns obtained from the siRNA screen.
Figure 4: Morphological focal adhesion characteristics and net contractile moments in PTK-knockdown cells with altered rigidity-dependent polarization.
Figure 5: Focal-adhesion-guided cell polarization, and its regulation by PTKs.

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Acknowledgements

The authors are grateful to B. Morgenstern for expert help in preparing this article for publication. This work was financially supported, in part, by the National Institutes of Health (NIH) Common Fund Nanomedicine Program (PN2 EY016586), the Israel Science Foundation, the Minerva Foundation, the Maurice Janin Fund and the De Benedetti Foundation-Cherasco. B.G. holds the Erwin Neter Professorial Chair in Cell and Tumor Biology. A.D.B. holds the Joseph Moss Professorial Chair in Biomedical Research.

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  1. Masha Prager-Khoutorsky, Ramaswamy Krishnan & Kavitha Rajendran

    Present address: Present addresses: Centre for Research in Neuroscience, McGill University, Montreal QC, H3G1A4, Canada (M.P-K.); Center for Vascular Biology Research, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA (R.K., K.R.),

  2. Masha Prager-Khoutorsky and Alexandra Lichtenstein: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel

    Masha Prager-Khoutorsky, Alexandra Lichtenstein, Avi Mayo, Zvi Kam, Benjamin Geiger & Alexander D. Bershadsky

  2. Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts 02115, USA

    Ramaswamy Krishnan & Kavitha Rajendran

  3. Mechanobiology Institute, National University of Singapore, 117411, Singapore

    Alexander D. Bershadsky

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  1. Masha Prager-Khoutorsky
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Contributions

M.P-K. and A.L. participated in the design of the experiments, carried them out and analysed the data. R.K. and K.R. carried out the traction force microscopy analysis. A.M. and Z.K. contributed to the microscopy analysis, image processing and statistical analysis of the data. A.D.B. and B.G. supervised the project and participated in all of its aspects, including its design, data analysis and manuscript preparation.

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Correspondence to Benjamin Geiger or Alexander D. Bershadsky.

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Prager-Khoutorsky, M., Lichtenstein, A., Krishnan, R. et al. Fibroblast polarization is a matrix-rigidity-dependent process controlled by focal adhesion mechanosensing. Nat Cell Biol 13, 1457–1465 (2011). https://doi.org/10.1038/ncb2370

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