Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Dynamic force patterns promote collective cell movements during embryonic wound repair

Nature Physics volume 14, pages 750–758 (2018)Cite this article

Subjects

Abstract

Embryonic wounds heal rapidly, in a process driven by coordinated cell movements. Polarization of actin and the molecular motor non-muscle myosin II in the cells adjacent to the wound results in the formation of a supracellular cable around the wound that drives repair. In Drosophila embryos, the distribution of actin around wounds is heterogeneous, with regions of high and low actin density, and actin heterogeneity is necessary for rapid repair. Here, we demonstrate that actin and myosin display stochastic patterns around embryonic wounds, and that contractile forces around wounds are heterogeneous. Mathematical modelling suggests that actomyosin heterogeneity favours wound closure if myosin is regulated by tension and strain, a hypothesis that we validate experimentally. We show that inhibition of stretch-activated ion channels disrupts myosin dynamics and tissue repair. Together, our results indicate that staggered contractile events, mechanical signals and force-regulated myosin dynamics coordinate cell behaviours to drive efficient wound closure.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Actin and myosin display similar, non-uniform distributions during embryonic wound repair.
Fig. 2: Heterogeneous actomyosin patterns result in non-uniform contraction.
Fig. 3: Myosin is randomly distributed at the wound edge.
Fig. 4: Mechanical regulation of myosin dynamics drives rapid wound repair in silico.
Fig. 5: Local tensile strain is sufficient for myosin localization to the cortex.
Fig. 6: SAICs are required for myosin dynamics during wound repair.

Similar content being viewed by others

References

  1. 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).

    Google Scholar 

  2. Fernandez-Gonzalez, R., Simoes Sde, M., Roper, J. C., Eaton, S. & Zallen, J. A. Myosin II dynamics are regulated by tension in intercalating cells. Dev. Cell 17, 736–743 (2009).

    Google Scholar 

  3. Lau, K. et al. Anisotropic stress orients remodelling of mammalian limb bud ectoderm. Nat. Cell Biol. 17, 569–579 (2015).

    Google Scholar 

  4. Rozbicki, E. et al. Myosin-II-mediated cell shape changes and cell intercalation contribute to primitive streak formation. Nat. Cell Biol. 17, 397–408 (2015).

    Google Scholar 

  5. Williams-Masson, E. M., Malik, A. N. & Hardin, J. An actin-mediated two-step mechanism is required for ventral enclosure of the C. elegans hypodermis. Development 124, 2889–2901 (1997).

    Google Scholar 

  6. Kiehart, D. P., Galbraith, C. G., Edwards, K. A., Rickoll, W. L. & Montague, R. A. Multiple forces contribute to cell sheet morphogenesis for dorsal closure in Drosophila. J. Cell Biol. 149, 471–490 (2000).

    Google Scholar 

  7. Martin, P. & Lewis, J. Actin cables and epidermal movement in embryonic wound healing. Nature 360, 179–183 (1992).

    Article  ADS  Google Scholar 

  8. Wood, W. et al. Wound healing recapitulates morphogenesis in Drosophila embryos. Nat. Cell Biol. 4, 907–912 (2002).

    Google Scholar 

  9. Davidson, L., Ezin, A. M. & Keller, R. Embryonic wound healing by apical contraction and ingression in Xenopus laevis. Cell Motil. Cytoskelet. 53, 163–176 (2002).

    Google Scholar 

  10. McCluskey, J., Hopkinson-Woolley, J., Luke, B. & Martin, P. A study of wound healing in the E11.5 mouse embryo by light and electron microscopy. Tissue Cell 25, 173–181 (1993).

    Google Scholar 

  11. Ducuing, A. & Vincent, S. The actin cable is dispensable in directing dorsal closure dynamics but neutralizes mechanical stress to prevent scarring in the Drosophila embryo. Nat. Cell Biol. 18, 1149–1160 (2016).

    Google Scholar 

  12. Brock, J., Midwinter, K., Lewis, J. & Martin, P. Healing of incisional wounds in the embryonic chick wing bud: characterization of the actin purse-string and demonstration of a requirement for Rho activation. J. Cell Biol. 135, 1097–1107 (1996).

    Google Scholar 

  13. Abreu-Blanco, M. T., Verboon, J. M. & Parkhurst, S. M. Drosophila embryos close epithelial wounds using a combination of cellular protrusions and an actomyosin purse string. J. Cell Sci. 125, 5984–5997 (2012).

    Google Scholar 

  14. Fernandez-Gonzalez, R. & Zallen, J. A. Wounded cells drive rapid epidermal repair in the early Drosophila embryo. Mol. Biol. Cell 24, 3227–3237 (2013).

    Google Scholar 

  15. Zulueta-Coarasa, T., Tamada, M., Lee, E. J. & Fernandez-Gonzalez, R. Automated multidimensional image analysis reveals a role for Abl in embryonic wound repair. Development 141, 2901–2911 (2014).

    Google Scholar 

  16. Bement, W. M., Forscher, P. & Mooseker, M. S. A novel cytoskeletal structure involved in purse string wound closure and cell polarity maintenance. J. Cell Biol. 121, 565–578 (1993).

    Google Scholar 

  17. Brugués, A. et al. Forces driving epithelial wound healing. Nat. Phys. 10, 683–690 (2014).

    Google Scholar 

  18. Kiehart, D. P. & Feghali, R. Cytoplasmic myosin from Drosophila melanogaster. J. Cell Biol. 103, 1517–1525 (1986).

    Google Scholar 

  19. Martin, A. C., Kaschube, M. & Wieschaus, E. F. Pulsed contractions of an actin-myosin network drive apical constriction. Nature 457, 495–499 (2009).

    ADS  Google Scholar 

  20. Hutson, M. S. et al. Forces for morphogenesis investigated with laser microsurgery and quantitative modeling. Science 300, 145–149 (2003).

    ADS  Google Scholar 

  21. Kumar, S. et al. Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics. Biophys. J. 90, 3762–3773 (2006).

    ADS  Google Scholar 

  22. Kovacs, M., Thirumurugan, K., Knight, P. J. & Sellers, J. R. Load-dependent mechanism of nonmuscle myosin 2. Proc. Natl Acad. Sci. USA 104, 9994–9999 (2007).

    ADS  Google Scholar 

  23. Kobb, A. B., Zulueta-Coarasa, T. & Fernandez-Gonzalez, R. Tension regulates myosin dynamics during Drosophila embryonic wound repair. J. Cell Sci. 130, 689–696 (2017).

    Google Scholar 

  24. Effler, J. C. et al. Mitosis-specific mechanosensing and contractile-protein redistribution control cell shape. Curr. Biol. 16, 1962–1967 (2006).

    Google Scholar 

  25. Yu, J. C. & Fernandez-Gonzalez, R. Local mechanical forces promote polarized junctional assembly and axis elongation in Drosophila. eLife 5, e10757 (2016).

    Google Scholar 

  26. Bowman, C. L., Gottlieb, P. A., Suchyna, T. M., Murphy, Y. K. & Sachs, F. Mechanosensitive ion channels and the peptide inhibitor GsMTx-4: history, properties, mechanisms and pharmacology. Toxicon 49, 249–270 (2007).

    Google Scholar 

  27. Yang, X. C. & Sachs, F. Block of stretch-activated ion channels in Xenopus oocytes by gadolinium and calcium ions. Science 243, 1068–1071 (1989).

    ADS  Google Scholar 

  28. Bardet, P. L. et al. A fluorescent reporter of caspase activity for live imaging. Proc. Natl Acad. Sci. USA 105, 13901–13905 (2008).

    ADS  Google Scholar 

  29. Hashimoto, H., Robin, F. B., Sherrard, K. M. & Munro, E. M. Sequential contraction and exchange of apical junctions drives zippering and neural tube closure in a simple chordate. Dev. Cell 32, 241–255 (2015).

    Google Scholar 

  30. Maddox, A. S., Lewellyn, L., Desai, A. & Oegema, K. Anillin and the septins promote asymmetric ingression of the cytokinetic furrow. Dev. Cell 12, 827–835 (2007).

    Google Scholar 

  31. Niisato, N., Ohta, M., Eaton, D. C. & Marunaka, Y. Hypotonic stress upregulates beta- and gamma-ENaC expression through suppression of ERK by inducing MKP-1. Am. J. Physiol. Ren. Physiol. 303, F240–252 (2012).

    Google Scholar 

  32. Antunes, M., Pereira, T., Cordeiro, J. V., Almeida, L. & Jacinto, A. Coordinated waves of actomyosin flow and apical cell constriction immediately after wounding. J. Cell Biol. 202, 365–379 (2013).

    Google Scholar 

  33. Xu, S. & Chisholm, A. D. A Gαq-Ca2+. signaling pathway promotes actin-mediated epidermal wound closure in C. elegans. Curr. Biol. 21, 1960–1967 (2011).

    Google Scholar 

  34. Hathaway, D. R. & Adelstein, R. S. Human platelet myosin light chain kinase requires the calcium-binding protein calmodulin for activity. Proc. Natl Acad. Sci. USA 76, 1653–1657 (1979).

    ADS  Google Scholar 

  35. Hartwig, J. H. et al. MARCKS is an actin filament crosslinking protein regulated by protein kinase C and calcium-calmodulin. Nature 356, 618–622 (1992).

    ADS  Google Scholar 

  36. Masiero, L., Lapidos, K. A., Ambudkar, I. & Kohn, E. C. Regulation of the RhoA pathway in human endothelial cell spreading on type IV collagen: role of calcium influx. J. Cell Sci. 112, 3205–3213 (1999).

    Google Scholar 

  37. Razzell, W., Evans, I. R., Martin, P. & Wood, W. Calcium flashes orchestrate the wound inflammatory response through DUOX activation and hydrogen peroxide release. Curr. Biol. 23, 424–429 (2013).

    Google Scholar 

  38. Hunter, M. V., Lee, D. M., Harris, T. J. & Fernandez-Gonzalez, R. Polarized E-cadherin endocytosis directs actomyosin remodeling during embryonic wound repair. J. Cell Biol. 210, 801–816 (2015).

    Google Scholar 

  39. Hunter, G. L., Crawford, J. M., Genkins, J. Z. & Kiehart, D. P. Ion channels contribute to the regulation of cell sheet forces during Drosophila dorsal closure. Development 141, 325–334 (2014).

    Google Scholar 

  40. Coste, B. et al. Piezo proteins are pore-forming subunits of mechanically activated channels. Nature 483, 176–181 (2012).

    ADS  Google Scholar 

  41. Gudipaty, S. A. et al. Mechanical stretch triggers rapid epithelial cell division through Piezo1. Nature 543, 118–121 (2017).

    ADS  Google Scholar 

  42. Zhao, P. Y. et al. TRP channels localize to subdomains of the apical plasma membrane in human fetal retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 56, 1916–1923 (2015).

    Google Scholar 

  43. Simone, R. P. & DiNardo, S. Actomyosin contractility and Discs large contribute to junctional conversion in guiding cell alignment within the Drosophila embryonic epithelium. Development 137, 1385–1394 (2010).

    Google Scholar 

  44. Marcinkevicius, E. & Zallen, J. A. Regulation of cytoskeletal organization and junctional remodeling by the atypical cadherin Fat. Development 140, 433–443 (2013).

    Google Scholar 

  45. Tan, P. Y. & Zaidel-Bar, R. Transient membrane localization of SPV-1 drives cyclical actomyosin contractions in the C. elegans spermatheca. Curr. Biol. 25, 141–151 (2015).

    Google Scholar 

  46. Royou, A., Field, C., Sisson, J. C., Sullivan, W. & Karess, R. Reassessing the role and dynamics of nonmuscle myosin II during furrow formation in early Drosophila embryos. Mol. Biol. Cell 15, 838–850 (2004).

    Google Scholar 

  47. Huang, J., Zhou, W. K., Dong, W., Watson, A. M. & Hong, Y. Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering. Proc. Natl Acad. Sci. USA 106, 8284–8289 (2009).

    ADS  Google Scholar 

  48. Oda, H. & Tsukita, S. Real-time imaging of cell-cell adherens junctions reveals that Drosophila mesoderm invagination begins with two phases of apical constriction of cells. J. Cell Sci. 114, 493–501 (2001).

    Google Scholar 

  49. Chen, T. W. et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013).

    ADS  Google Scholar 

  50. Foe, V. E. & Alberts, B. M. Studies of nuclear and cytoplasmic behavior during the 5 mitotic-cycles that precede gastrulation in Drosophila embryogenesis. J. Cell Sci. 61, 31–70 (1983).

    Google Scholar 

  51. Kiehart, D. P., Crawford, J. M. & Montague, R. A. Quantitative microinjection of Drosophila embryos: general strategy. CSH Protoc. 2007, pdbtop5 (2007).

    Google Scholar 

  52. Nishizawa, M. & Nishizawa, K. Molecular dynamics simulations of a stretch-activated channel inhibitor GsMTx4 with lipid membranes: two binding modes and effects of lipid structure. Biophys. J. 92, 4233–4243 (2007).

    ADS  Google Scholar 

  53. Hurst, A. C., Gottlieb, P. A. & Martinac, B. Concentration dependent effect of GsMTx4 on mechanosensitive channels of small conductance in E. coli spheroplasts. Eur. Biophys. J. 38, 415–425 (2009).

    Google Scholar 

  54. Fernandez-Gonzalez, R. & Zallen, J. A. Oscillatory behaviors and hierarchical assembly of contractile structures in intercalating cells. Phys. Biol. 8, 045005 (2011).

    ADS  Google Scholar 

  55. Leung, C. Y. & Fernandez-Gonzalez, R. Quantitative image analysis of cell behavior and molecular dynamics during tissue morphogenesis. Methods Mol. Biol. 1189, 99–113 (2015).

    Google Scholar 

  56. Fletcher, A. G., Osborne, J. M., Maini, P. K. & Gavaghan, D. J. Implementing vertex dynamics models of cell populations in biology within a consistent computational framework. Prog. Biophys. Mol. Biol. 113, 299–326 (2013).

    Google Scholar 

  57. Yu, J. C. & Fernandez-Gonzalez, R. Quantitative modelling of epithelial morphogenesis: integrating cell mechanics and molecular dynamics. Semin. Cell Dev. Biol. 67, 153–160 (2016).

    Google Scholar 

  58. Razzell, W., Wood, W. & Martin, P. Recapitulation of morphogenetic cell shape changes enables wound re-epithelialisation. Development 141, 1814–1820 (2014).

    Google Scholar 

  59. Lan, H., Wang, Q., Fernandez-Gonzalez, R. & Feng, J. J. A biomechanical model for cell polarization and intercalation during Drosophila germband extension. Phys. Biol. 12, 056011 (2015).

    ADS  Google Scholar 

  60. Merkel, R. et al. A micromechanic study of cell polarity and plasma membrane cell body coupling in Dictyostelium. Biophys. J. 79, 707–719 (2000).

    ADS  Google Scholar 

  61. Ren, Y. X. et al. Mechanosensing through cooperative interactions between myosin II and the actin crosslinker cortexillin I. Curr. Biol. 19, 1421–1428 (2009).

    Google Scholar 

  62. Koride, S. et al. Mechanochemical regulation of oscillatory follicle cell dynamics in the developing Drosophila egg chamber. Mol. Biol. Cell 25, 3709–3716 (2014).

    Google Scholar 

  63. Bambardekar, K., Clement, R., Blanc, O., Chardes, C. & Lenne, P. F. Direct laser manipulation reveals the mechanics of cell contacts in vivo. Proc. Natl Acad. Sci. USA 112, 1416–1421 (2015).

    ADS  Google Scholar 

  64. Glantz, S. A. Primer of Biostatistics (McGraw-Hill, New York, 2002).

Download references

Acknowledgements

We are grateful to T. Harris for sharing the myosin II antibody, to M. Hunter and J. Yu for technical assistance, and to C. Tong for help with data annotation. We thank A. McGuigan, C. Simmons and R. Winklbauer for useful discussions; and J. Feng, S. Hopyan, M. Hunter, A. Kobb, C. McFaul and J. Yu for comments on the manuscript. Flybase provided important information for this study. T.Z.-C. was supported by an Ontario Trillium Scholarship and a Doctoral Completion Award from the University of Toronto, and is a World Fellow of the Delta Kappa Gamma International Society. This work was supported by grants to R.F.-G. from the Natural Sciences and Engineering Research Council of Canada (418438-13), the Canada Foundation for Innovation (30279), the Ontario Ministry of Economic Development and Innovation (ER14-10-170), the Ted Rogers Centre for Heart Research TBEP Seed Program, and the Canada First Research Excellence Fund-University of Toronto Medicine by Design. R.F.-G. is the Tier II Canada Research Chair in Quantitative Cell Biology and Morphogenesis.

Author information

Authors and Affiliations

  1. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

    Teresa Zulueta-Coarasa & Rodrigo Fernandez-Gonzalez

  2. Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto, Toronto, Ontario, Canada

    Teresa Zulueta-Coarasa & Rodrigo Fernandez-Gonzalez

  3. Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario, Canada

    Rodrigo Fernandez-Gonzalez

  4. Developmental and Stem Cell Biology Program, The Hospital for Sick Children, Toronto, Ontario, Canada

    Rodrigo Fernandez-Gonzalez

Authors
  1. Teresa Zulueta-Coarasa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rodrigo Fernandez-Gonzalez
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

T.Z.-C. and R.F.-G. developed the initial ideas for this study, analysed the data and prepared the manuscript. T.Z.-C. performed all experiments.

Corresponding author

Correspondence to Rodrigo Fernandez-Gonzalez.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Materials

Supplementary Figures 1–15, Supplementary Tables 1 and 2, Supplementary Video Legends 1–10, Supplementary References 1–3

Reporting Summary

Supplementary Video 1

Actin and myosin display similar patterns at the wound margin. Epidermal wound in an embryo co-expressing GFP:MoesinABD and myosin:mCherry. A stack was acquired every 30 s for 1 h and 39 min. Time after wounding is shown. Anterior left, dorsal up.

Supplementary Video 2

Mechanical forces at the wound margin are heterogeneous. Laser ablation experiments in embryos expressing E-cadherin:GFP. Cuts were performed in segments of the actomyosin cable with high (left) or low (right) myosin levels (see Fig. 2a,b). Yellow Xs indicate the sites of ablation. A stack was acquired every 3 s for 66 s. Time after ablation is shown. Anterior left, dorsal up.

Supplementary Video 3

Contraction is faster when the actomyosin distribution is heterogeneous. Heterogeneous (left) and homogeneous (right) patches of the wound margin in embryos expressing E-cadherin:mTomato (green) and myosin:GFP (magenta). Yellow arrowheads indicate the tracked segment. A stack was acquired every 10 s for 1 min.

Supplementary Video 4

Heterogeneous myosin distribution and mechanically regulated dynamics favour rapid wound closure in silico. Wound healing simulations with a heterogeneous myosin distribution and no myosin turnover (left), with tension-based myosin stabilization (centre-left), with strain-based myosin recruitment (centre), or with strain and tension-dependent myosin dynamics (centre-right); or with a homogeneous myosin distribution and strain and tension dependent myosin dynamics (right). Colour indicates interfacial tension in nN based on myosin levels (see Fig. 4 for scale). The simulation time step was 2 s for 20 min. Time is with respect to the onset of wound closure. Anterior left, dorsal up.

Supplementary Video 5

Non-uniform wound edge deformation is associated with embryonic wound repair. Strain maps of the wound edge during embryonic repair. Colour indicates the degree and the type of strain (tensile, cold colours; compressive, warm colours). A stack was acquired every 10 s for 20.3 min. Time after wounding is shown. Anterior left, dorsal up.

Supplementary Video 6

Myosin accumulates at a wound edge segment after being stretched. Relative timing between the deformation of wound edge segment and recruitment of myosin in an embryo expressing E-cadherin:mTomato (green) and myosin:GFP (magenta). Yellow arrowheads indicate the segment of interest. A stack was acquired every 10 s for 20.3 min. Time after wounding is shown. Anterior left, dorsal up.

Supplementary Video 7

Segments mechanically isolated from their neighbours fail to recruit myosin. Wound edge segments in embryos expressing E-cadherin:mTomato (green) and myosin:GFP (magenta) far from the site of wound edge ablation (left) or in a mechanically isolated segment (right). Yellow arrowheads indicate the segments in which fluorescence was monitored. Cyan Xs indicate the segments targeted for ablation. A stack was acquired every 30 s for 20.5 min. Time after segment isolation is shown. Anterior left, dorsal up.

Supplementary Video 8

Tensile strain is sufficient to recruit myosin. Cell interfaces in embryos expressing E-cadherin:mTomato (green) and myosin:GFP (magenta) in sham (left) or UV-irradiated 32 embryos (right). Yellow arrowheads indicate the monitored interfaces. Cyan Xs indicate the cells irradiated to induce strain. A stack was acquired every 10 s for 20.3 min. Time after wounding is shown. Anterior left, dorsal up.

Supplementary Video 9

GsMTx4 treatment impairs wound closure. Wounds in embryos expressing myosin:GFP and injected with buffer (left) or 2.5 mM GsMTx4 (right). A stack was acquired every 30 s for 39 min. Time after injection is shown. Anterior left, dorsal up.

Supplementary Video 10

GdCl3 injection prevents wound repair. Wounds in embryos expressing myosin:GFP and injected with water (left) or 20 mM GdCl3 (right). A stack was acquired every 30 s for 39.5 min. Time after injection is shown. Anterior left, dorsal up.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zulueta-Coarasa, T., Fernandez-Gonzalez, R. Dynamic force patterns promote collective cell movements during embryonic wound repair. Nature Phys 14, 750–758 (2018). https://doi.org/10.1038/s41567-018-0111-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41567-018-0111-2

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing