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Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates

Nature Cell Biology volume 3, pages 466–472 (2001)Cite this article

Abstract

Mechanical forces play a major role in the regulation of cell adhesion and cytoskeletal organization. In order to explore the molecular mechanism underlying this regulation, we have investigated the relationship between local force applied by the cell to the substrate and the assembly of focal adhesions. A novel approach was developed for real-time, high-resolution measurements of forces applied by cells at single adhesion sites. This method combines micropatterning of elastomer substrates and fluorescence imaging of focal adhesions in live cells expressing GFP-tagged vinculin. Local forces are correlated with the orientation, total fluorescence intensity and area of the focal adhesions, indicating a constant stress of 5.5 ± 2 nNμm-2. The dynamics of the force-dependent modulation of focal adhesions were characterized by blocking actomyosin contractility and were found to be on a time scale of seconds. The results put clear constraints on the possible molecular mechanisms for the mechanosensory response of focal adhesions to applied force.

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Figure 1: Fabrication and calibration of the patterned elastomer.
Figure 2: Cells plated on the patterned elastomer create distortions.
Figure 3: Visualization of forces and focal adhesions.
Figure 4: Distribution of forces in a cardiac myocyte.
Figure 5: Correlation between force and focal adhesion structure.

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Acknowledgements

We wish to thank E. Zamir and E. Moses for illuminating discussions and technical assistance. This study was supported by the Israel Science Foundation, administrated by the Israel Academy of Science and the Minerva Foundation. B.G. holds the E. Neter chair for Cell and Tumor Biology. L.A. is incumbent of the Dorothy and Patrick Gorman Professorial chair of Biological Ultrastructure. U.S.S. was supported by the Minerva Foundation. A. B. holds the J. Moss chair of Biomedical Research.

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Authors and Affiliations

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

    Nathalie Q. Balaban, Polina Goichberg, Gila Tzur, Ilana Sabanay, Alexander Bershadsky & Benjamin Geiger

  2. Department of Materials and Interfaces, The Weizmann Institute of Science, Rehovot, Israel

    Ulrich S. Schwarz & Sam Safran

  3. Laboratoire de Spectrometrie Physique, Universite Joseph Fourier, Grenoble, France

    Daniel Riveline

  4. Braun Center for Submicron Semiconductor Research, The Weizmann Institute of Science, Rehovot, Israel

    Diana Mahalu

  5. Department of Structural Biology, The Weizmann Institute of Science, Rehovot, Israel

    Lia Addadi

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  1. Nathalie Q. Balaban
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Correspondence to Benjamin Geiger.

Supplementary information

Movie 1

Two beating cardiac myocytes on the micropatterned elastomer. Beating is reconstructed from pictures acquired either in the contracted or the relaxed phase. The border between patterned and non-patterned elastomer can be seen in the lower part of the picture. (MOV 292 kb)

Movie 2

A cardiac myocyte plated on the micropatterned elastic substrate. Reconstruction from pictures acquired sequentially either in the relaxed or the contracted phase. White crosses indicate regions of contraction; black cross in the upper left shows a non-disturbed location. (MOV 809 kb)

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Balaban, N., Schwarz, U., Riveline, D. et al. Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates. Nat Cell Biol 3, 466–472 (2001). https://doi.org/10.1038/35074532

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