Epithelial monolayers are one-cell-thick tissue sheets that line most of the body surfaces, separating internal and external environments. As part of their function, they must withstand extrinsic mechanical stresses applied at high strain rates. However, little is known about how monolayers respond to mechanical deformations. Here, by subjecting suspended epithelial monolayers to stretch, we find that they dissipate stresses on a minute timescale and that relaxation can be described by a power law with an exponential cut-off at timescales larger than about 10 s. This process involves an increase in monolayer length, pointing to active remodelling of cellular biopolymers at the molecular scale during relaxation. Strikingly, monolayers consisting of tens of thousands of cells relax stress with similar dynamics to single rounded cells, and both respond similarly to perturbations of the actomyosin cytoskeleton. By contrast, cell–cell junctional complexes and intermediate filaments do not relax tissue stress, but form stable connections between cells, allowing monolayers to behave rheologically as single cells. Taken together, our data show that actomyosin dynamics governs the rheological properties of epithelial monolayers, dissipating applied stresses and enabling changes in monolayer length.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
The Effect of Substrate Stiffness on Elastic Force Transmission in the Epithelial Monolayers over Short Timescales
Cellular and Molecular Bioengineering Open Access 13 July 2023
Nature Communications Open Access 17 August 2022
A hierarchical cellular structural model to unravel the universal power-law rheological behavior of living cells
Nature Communications Open Access 18 October 2021
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
All data supporting the conclusions are available from the authors on reasonable request.
Custom-written code used for data analysis is available from the authors on request.
Heisenberg, C.-P. & Bellaïche, Y. Forces in tissue morphogenesis and patterning. Cell 153, 948–962 (2013).
Martin, A. C., Gelbart, M., Fernandez-Gonzalez, R., Kaschube, M. & Wieschaus, E. F. Integration of contractile forces during tissue invagination. J. Cell Biol. 188, 735–749 (2010).
Tschumperlin, D. J., Boudreault, F. & Liu, F. Recent advances and new opportunities in lung mechanobiology. J. Biomech. 43, 99 (2010).
Califano, J. P. & Reinhart-King, C. A. Exogenous and endogenous force regulation of endothelial cell behavior. J. Biomech. 43, 79–86 (2010).
Blanchard, G. B. et al. Tissue tectonics: morphogenetic strain rates, cell shape change and intercalation. Nat. Methods 6, 458–464 (2009).
He, Z., Ritchie, J., Grashow, J. S., Sacks, M. S. & Yoganathan, A. P. In vitro dynamic strain behavior of the mitral valve posterior leaflet. J. Biomech. Eng. 127, 504–511 (2005).
Sacks, M. S. et al. In-vivo dynamic deformation of the mitral valve anterior leaflet. Ann. Thorac. Surg. 82, 1369–1377 (2006).
Perlman, C. E. & Bhattacharya, J. Alveolar expansion imaged by optical sectioning microscopy. J. Appl. Physiol. 103, 1037–1044 (2007).
Padala, M. et al. Mechanics of the mitral valve strut chordae insertion region. J. Biomech. Eng. 132, 081004 (2010).
Maiti, R. et al. In vivo measurement of skin surface strain and sub-surface layer deformation induced by natural tissue stretching. J. Mech. Behav. Biomed. Mater. 62, 556–569 (2016).
Korkmaz, I. & Rogg, B. A simple fluid-mechanical model for the prediction of the stress–strain relation of the male urinary bladder. J. Biomech. 40, 663–668 (2007).
Obropta, E. W. & Newman, D. J. Skin strain fields at the shoulder joint for mechanical counter pressure space suit development. In 2016 IEEE Aerospace Conference 2562–2570 (IEEE, 2016).
Harris, A. R. et al. Characterizing the mechanics of cultured cell monolayers. Proc. Natl Acad. Sci. USA 109, 16449–16454 (2012).
Suki, B. & Hubmayr, R. Epithelial and endothelial damage induced by mechanical ventilation modes. Curr. Opin. Crit. Care 20, 17–24 (2014).
Jufri, N. F., Mohamedali, A., Avolio, A. & Baker, M. S. Mechanical stretch: physiological and pathological implications for human vascular endothelial cells. Vasc. Cell 7, 8 (2015).
Getsios, S., Huen, A. C. & Green, K. J. Working out the strength and flexibility of desmosomes. Nat. Rev. Mol. Cell Biol. 5, 271–281 (2004).
Levine, E., Lee, C. H., Kintner, C. & Gumbiner, B. M. Selective disruption of E-cadherin function in early Xenopus embryos by a dominant negative mutant. Development 120, 901–909 (1994).
Tang, V. W. & Brieher, W. M. FSGS3/CD2AP is a barbed-end capping protein that stabilizes actin and strengthens adherens junctions. J. Cell Biol. 203, 815–833 (2013).
Wyatt, T., Baum, B. & Charras, G. A question of time: tissue adaptation to mechanical forces. Curr. Opin. Cell Biol. 38, 68–73 (2016).
Moeendarbary, E. et al. The cytoplasm of living cells behaves as a poroelastic material. Nat. Mater. 12, 253–261 (2013).
Trepat, X. et al. Universal physical responses to stretch in the living cell. Nature 447, 592–595 (2007).
Fischer-Friedrich, E. et al. Rheology of the active cell cortex in mitosis. Biophys. J. 111, 589–600 (2016).
Priya, R. et al. Feedback regulation through myosin II confers robustness on RhoA signalling at E-cadherin junctions. Nat. Cell Biol. 17, 1282–1293 (2015).
Charras, G. & Yap, A. S. Tensile forces and mechanotransduction at cell–cell junctions. Curr. Biol. 28, R445–R457 (2018).
Bambardekar, K., Clément, R., Blanc, O., Chardès, C. & Lenne, P.-F. Direct laser manipulation reveals the mechanics of cell contacts in vivo. Proc. Natl Acad. Sci. USA 112, 1416–1421 (2015).
Harris, A. R. et al. Generating suspended cell monolayers for mechanobiological studies. Nat. Protoc. 8, 2516–2530 (2013).
Roan, E. & Waters, C. M. What do we know about mechanical strain in lung alveoli? Am. J. Physiol.—Lung Cell. Mol. Physiol. 301, L625–L635 (2011).
Wyatt, T. P. J. et al. Emergence of homeostatic epithelial packing and stress dissipation through divisions oriented along the long cell axis. Proc. Natl Acad. Sci. USA 112, 5726–5731 (2015).
Lecuit, T. & Yap, A. S. E-cadherin junctions as active mechanical integrators in tissue dynamics. Nat. Cell Biol. 17, 533–539 (2015).
Forgacs, G., Foty, R. A., Shafrir, Y. & Steinberg, M. S. Viscoelastic properties of living embryonic tissues: a quantitative study. Biophys. J. 74, 2227–2234 (1998).
Kollmannsberger, P. & Fabry, B. Linear and nonlinear rheology of living cells. Annu. Rev. Mater. Res. 41, 75–97 (2011).
Khalilgharibi, N., Fouchard, J., Recho, P., Charras, G. & Kabla, A. The dynamic mechanical properties of cellularised aggregates. Curr. Opin. Cell Biol. 42, 113–120 (2016).
Bonnet, I. et al. Mechanical state, material properties and continuous description of an epithelial tissue. J. R. Soc. Interface 9, 2614–2623 (2012).
Martin, A. C., Kaschube, M. & Wieschaus, E. F. Pulsed actin-myosin network contractions drive apical constriction. Nature 457, 495 (2009).
Machado, P. F. et al. Emergent material properties of developing epithelial tissues. BMC Biol. 13, 98 (2015).
Wang, N. & Stamenović, D. Contribution of intermediate filaments to cell stiffness, stiffening, and growth. Am. J. Physiol.—Cell Physiol. 279, C188 (2000).
Ramms, L. et al. Keratins as the main component for the mechanical integrity of keratinocytes. Proc. Natl Acad. Sci. USA 110, 18513–18518 (2013).
Harris, A., Daeden, A. & Charras, G. Formation of adherens junctions leads to the emergence of a tissue-level tension in epithelial monolayers. J. Cell Sci. 127, 2507–2517 (2014).
Gonzalez-Rodriguez, D. et al. Detachment and fracture of cellular aggregates. Soft Matter 9, 2282–2290 (2013).
Cavey, M., Rauzi, M., Lenne, P.-F. & Lecuit, T. A two-tiered mechanism for stabilization and immobilization of E-cadherin. Nature 453, 751–756 (2008).
Kovacs, E. M. et al. N-WASP regulates the epithelial junctional actin cytoskeleton through a non-canonical post-nucleation pathway. Nat. Cell Biol. 13, 934 (2011).
Yoshinaga, N. & Marcq, P. Contraction of cross-linked actomyosin bundles. Phys. Biol. 9, 046004 (2012).
Desprat, N., Guiroy, A. & Asnacios, A. Microplates-based rheometer for a single living cell. Rev. Sci. Instrum. 77, 055111 (2006).
Fischer-Friedrich, E., Hyman, A. A., Jülicher, F., Müller, D. J. & Helenius, J. Quantification of surface tension and internal pressure generated by single mitotic cells. Sci. Rep. 4, 6213 (2014).
Muñoz, J. J. & Albo, S. Physiology-based model of cell viscoelasticity. Phys. Rev. E 88, 012708 (2013).
Doubrovinski, K., Swan, M., Polyakov, O. & Wieschaus, E. F. Measurement of cortical elasticity in Drosophila melanogaster embryos using ferrofluids. Proc. Natl Acad. Sci. USA 114, 1051–1056 (2017).
Clément, R., Dehapiot, B., Collinet, C., Lecuit, T. & Lenne, P.-F. Viscoelastic dissipation stabilizes cell shape changes during tissue morphogenesis. Curr. Biol. 27, 3132–3142 (2017).
Prost, J., Julicher, F. & Joanny, J. F. Active gel physics. Nat. Phys. 11, 111–117 (2015).
Salbreux, G., Charras, G. & Paluch, E. Actin cortex mechanics and cellular morphogenesis. Trends Cell Biol. 22, 536–545 (2012).
Biro, M. et al. Cell cortex composition and homeostasis resolved by integrating proteomics and quantitative imaging. Cytoskeleton 70, 741–754 (2013).
Chugh, P. et al. Actin cortex architecture regulates cell surface tension. Nat. Cell Biol. 19, 689–697 (2017).
Charras, G. T., Hu, C. K., Coughlin, M. & Mitchison, T. J. Reassembly of contractile actin cortex in cell blebs. J. Cell Biol. 175, 477–490 (2006).
Kajita, M. et al. Filamin acts as a key regulator in epithelial defence against transformed cells. Nat. Commun. 5, 4428 (2014).
The authors wish to acknowledge present and past members of the Charras, Baum, Kabla and Muñoz laboratories for discussions. The authors acknowledge technical support from UCL Genomics for sequencing and analysing total RNA data as well as from J. Duque (LCN) for analysis of fluorescence intensity at junctions and from M. Vaghela (LCN) for AFM measurements. N.K. was funded by the Rosetrees Trust, the UCL Graduate School, the EPSRC funded doctoral training programme CoMPLEX and the European Research Council (ERC-CoG MolCellTissMech, agreement 647186 to G.C.). N.K. was in receipt of a UCL Overseas Research Scholarship. N.K. was supported by the Professor Rob Seymour Travel Bursary Fund for research visits to Barcelona. J.F. and A.B. were funded by BBSRC grants (BB/M003280 and BB/M002578) to G.C. and A.K. J.J.M., N.A. and P.M. acknowledge the support of the Ministry of Economy, Industry and Competitiveness through grants nos. DPI2013-43727R and DPI2016-74929-R and the Generalitat de Catalunya through grant no. 2014-SGR-1471. N.A. was also financially supported by Universitat Politècnica de Catalunya and Consorci Escola Industrial de Barcelona through grant UPC-FPI 2012, and the European Research Council under the European Community’s Seventh Framework Programme (FP7/2007–2013)/ERC, grant agreement no. 240487. P.M. was also supported by the European Molecular and Biology Organization under grant ASTF 351-2016. R.B. is part of the EPSRC-funded doctoral training programme CoMPLEX. M.D. was funded by a Marie Skłodowska-Curie Horizon 2020 Individual Fellowship (MRTGS). A.Y. was supported by an HFSP Young Investigator award to G.C. (RGY 66/2013). A.H. was supported by a BBSRC grant (BB/K013521) to G.C. and A.K. Y.F. was supported by Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research on Innovative Areas 26114001, Grant-in-Aid for Scientific Research (A) 18H03994, the Strategic Japanese–Swiss Science and Technology Programme, AMED under grant nos. JP17ck0106361 and JP18cm0106234, SAN-ESU GIKEN Co. Ltd, the Naito Foundation and the Takeda Science Foundation. A.K. was supported by BBSRC grants (BB/K018175/1, BB/M003280 and BB/M002578). Y.M. is funded by MRC Fellowship MR/L009056/1, a UCL Excellence Fellowship and NSFC International Young Scientist Fellowship 31650110472. B.B. is supported by UCL, a BBSRC project grant (BB/K009001/1) and a CRUK programme grant (17343). M.M. is supported by EPSRC (EP/K038656/1). G.C. is supported by a consolidator grant from the European Research Council (MolCellTissMech, agreement 647186). Atomic force microscopy equipment was purchased thanks to an ALERT16 grant from BBSRC to G.C.
The authors declare no competing interests.
Journal peer review information: Nature Physics thanks Pierre-François Lenne and the other anonymous reviewer(s) for their contribution to the peer review of this work.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Figs. 1–20, Supplementary Tables 1–5, Supplementary Results, Supplementary Methods and Supplementary References 1–21.
Cross-section view of a monolayer expressing E-cadherin–GFP before, during and after stretch. The monolayer is stretched at 0 s and the strain is maintained constant at 30% until 129.6 s, after which the rod is returned to its initial position. The length of the monolayer on release is different from its length before application of stretch and, as a consequence, it buckles. The monolayer appears in green, the surrounding medium appears in magenta due to inclusion of Alexa-647 and the glass substrates on either side of the suspended monolayer appear dark due to dye exclusion. Scale bar: 100 μm.
About this article
Cite this article
Khalilgharibi, N., Fouchard, J., Asadipour, N. et al. Stress relaxation in epithelial monolayers is controlled by the actomyosin cortex. Nat. Phys. 15, 839–847 (2019). https://doi.org/10.1038/s41567-019-0516-6
This article is cited by
The Effect of Substrate Stiffness on Elastic Force Transmission in the Epithelial Monolayers over Short Timescales
Cellular and Molecular Bioengineering (2023)
Nature Communications (2022)
Acta Mechanica Sinica (2022)
Biomechanics and Modeling in Mechanobiology (2022)
European Biophysics Journal (2022)