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.

  • Letter
  • Published:

A role for Rho GTPases and cell–cell adhesion in single-cell motility in vivo

Nature Cell Biology volume 12, pages 47–53 (2010)Cite this article

Abstract

Cell migration is central to embryonic development, homeostasis and disease1, processes in which cells move as part of a group or individually. Whereas the mechanisms controlling single-cell migration in vitro are relatively well understood2,3,4, less is known about the mechanisms promoting the motility of individual cells in vivo. In particular, it is not clear how cells that form blebs in their migration use those protrusions to bring about movement in the context of the three-dimensional cellular environment5,6. Here we show that the motility of chemokine-guided germ cells within the zebrafish embryo requires the function of the small Rho GTPases Rac1 and RhoA, as well as E-cadherin-mediated cell–cell adhesion. Using fluorescence resonance energy transfer we demonstrate that Rac1 and RhoA are activated in the cell front. At this location, Rac1 is responsible for the formation of actin-rich structures, and RhoA promotes retrograde actin flow. We propose that these actin-rich structures undergoing retrograde flow are essential for the generation of E-cadherin-mediated traction forces between the germ cells and the surrounding tissue and are therefore crucial for cell motility in vivo.

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

Figure 1: Actin cytoskeleton dynamics and Rac activity and function during germ-cell migration.
Figure 2: RhoA activity and role in regulating actin cytoskeleton dynamics in migrating germ cells.
Figure 3: Cell–cell adhesion in germ-cell migration.
Figure 4: The role of E-cadherin in germ-cell migration.
Figure 5: A model for cooperation between blebbing, actin retrograde flow and E-cadherin-mediated traction force in germ-cell motility.

Similar content being viewed by others

References

  1. Horwitz, R. & Webb, D. Cell migration. Curr. Biol. 13, R756–R759 (2003).

    Article  CAS  Google Scholar 

  2. Lauffenburger, D. A. & Horwitz, A. F. Cell migration: a physically integrated molecular process. Cell 84, 359–369 (1996).

    Article  CAS  Google Scholar 

  3. Ridley, A. J. et al. Cell migration: integrating signals from front to back. Science 302, 1704–1709 (2003).

    Article  CAS  Google Scholar 

  4. Puklin-Faucher, E. & Sheetz, M. P. The mechanical integrin cycle. J. Cell Sci. 122, 179–186 (2009).

    Article  CAS  Google Scholar 

  5. Fackler, O. T. & Grosse, R. Cell motility through plasma membrane blebbing. J. Cell Biol. 181, 879–884 (2008).

    Article  CAS  Google Scholar 

  6. Charras, G. & Paluch, E. Blebs lead the way: how to migrate without lamellipodia. Nature Rev. Mol. Cell Biol. 9, 730–736 (2008).

    Article  CAS  Google Scholar 

  7. Kunwar, P. S., Siekhaus, D. E. & Lehmann, R. In vivo migration: a germ cell perspective. Annu. Rev. Cell Dev. Biol. 22, 237–265 (2006).

    Article  CAS  Google Scholar 

  8. Doitsidou, M. et al. Guidance of primordial germ cell migration by the chemokine SDF-1. Cell 111, 647–659 (2002).

    Article  CAS  Google Scholar 

  9. Reichman-Fried, M., Minina, S. & Raz, E. Autonomous modes of behavior in primordial germ cell migration. Dev. Cell 6, 589–596 (2004).

    Article  CAS  Google Scholar 

  10. Blaser, H. et al. Migration of zebrafish primordial germ cells: a role for myosin contraction and cytoplasmic flow. Dev. Cell 11, 613–627 (2006).

    Article  CAS  Google Scholar 

  11. Boldajipour, B. et al. Control of chemokine-guided cell migration by ligand sequestration. Cell 132, 463–473 (2008).

    Article  CAS  Google Scholar 

  12. Heasman, S. J. & Ridley, A. J. Mammalian Rho GTPases: new insights into their functions from in vivo studies. Nature Rev. Mol. Cell Biol. 9, 690–701 (2008).

    Article  CAS  Google Scholar 

  13. Jaffe, A. B. & Hall, A. Rho GTPases: biochemistry and biology. Annu. Rev. Cell Dev. Biol. 21, 247–269 (2005).

    Article  CAS  Google Scholar 

  14. Sanders, L. C., Matsumura, F., Bokoch, G. M. & de Lanerolle, P. Inhibition of myosin light chain kinase by p21-activated kinase. Science 283, 2083–2085 (1999).

    Article  CAS  Google Scholar 

  15. Itoh, R. E. et al. Activation of rac and cdc42 video imaged by fluorescent resonance energy transfer-based single-molecule probes in the membrane of living cells. Mol. Cell. Biol. 22, 6582–6591 (2002).

    Article  CAS  Google Scholar 

  16. Wong, K., Pertz, O., Hahn, K. & Bourne, H. Neutrophil polarization: spatiotemporal dynamics of RhoA activity support a self-organizing mechanism. Proc. Natl Acad. Sci. USA 103, 3639–3644 (2006).

    Article  CAS  Google Scholar 

  17. Sander, E. E., ten Klooster, J. P., van Delft, S., van der Kammen, R. A. & Collard, J. G. Rac downregulates Rho activity: reciprocal balance between both GTPases determines cellular morphology and migratory behavior. J. Cell Biol. 147, 1009–1022 (1999).

    Article  CAS  Google Scholar 

  18. Pertz, O., Hodgson, L., Klemke, R. L. & Hahn, K. M. Spatiotemporal dynamics of RhoA activity in migrating cells. Nature 440, 1069–1072 (2006).

    Article  CAS  Google Scholar 

  19. Kurokawa, K. & Matsuda, M. Localized RhoA activation as a requirement for the induction of membrane ruffling. Mol. Biol. Cell 16, 4294–4303 (2005).

    Article  CAS  Google Scholar 

  20. Yoshizaki, H. et al. Activity of Rho-family GTPases during cell division as visualized with FRET-based probes. J. Cell Biol. 162, 223–232 (2003).

    Article  CAS  Google Scholar 

  21. Gupton, S. L. & Waterman-Storer, C. M. Spatiotemporal feedback between actomyosin and focal-adhesion systems optimizes rapid cell migration. Cell 125, 1361–1374 (2006).

    Article  CAS  Google Scholar 

  22. Gardel, M. L. et al. Traction stress in focal adhesions correlates biphasically with actin retrograde flow speed. J. Cell Biol. 183, 999–1005 (2008).

    Article  CAS  Google Scholar 

  23. Mould, A. P. et al. Identification of multiple integrin β1 homologs in zebrafish (Danio rerio). BMC Cell Biol. 7, 24 (2006).

    Article  Google Scholar 

  24. Gumbiner, B. M. Regulation of cadherin-mediated adhesion in morphogenesis. Nature Rev. Mol. Cell Biol. 6, 622–634 (2005).

    Article  CAS  Google Scholar 

  25. Blaser, H. et al. Transition from non-motile behaviour to directed migration during early PGC development in zebrafish. J. Cell Sci. 118, 4027–4038 (2005).

    Article  CAS  Google Scholar 

  26. Mich, J. K. et al. Germ cell migration in zebrafish is cyclopamine-sensitive but Smoothened-independent. Dev. Biol. 328, 342–354 (2009).

    Article  CAS  Google Scholar 

  27. Kemler, R. From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. Trends Genet. 9, 317–321 (1993).

    Article  CAS  Google Scholar 

  28. Nelson, W. J. Regulation of cell–cell adhesion by the cadherin–catenin complex. Biochem. Soc. Trans. 36, 149–155 (2008).

    Article  CAS  Google Scholar 

  29. Oyama, T. et al. A truncated β-catenin disrupts the interaction between E-cadherin and α-catenin: a cause of loss of intercellular adhesiveness in human cancer cell lines. Cancer Res. 54, 6282–6287 (1994).

    CAS  PubMed  Google Scholar 

  30. Kunwar, P. S. et al. Tre1 GPCR initiates germ cell transepithelial migration by regulating Drosophila melanogaster E-cadherin. J. Cell Biol. 183, 157–168 (2008).

    Article  CAS  Google Scholar 

  31. Kane, D. A., McFarland, K. N. & Warga, R. M. Mutations in half baked/E-cadherin block cell behaviors that are necessary for teleost epiboly. Development 132, 1105–1116 (2005).

    Article  CAS  Google Scholar 

  32. Lin, F. et al. Gα12/13 regulate epiboly by inhibiting E-cadherin activity and modulating the actin cytoskeleton. J. Cell Biol. 184, 909–921 (2009).

    Article  CAS  Google Scholar 

  33. Geisbrecht, E. R. & Montell, D. J. Myosin VI is required for E-cadherin-mediated border cell migration. Nature Cell Biol. 4, 616–620 (2002).

    Article  CAS  Google Scholar 

  34. Fulga, T. A. & Rorth, P. Invasive cell migration is initiated by guided growth of long cellular extensions. Nature Cell Biol. 4, 715–719 (2002).

    Article  CAS  Google Scholar 

  35. Hogan, C. et al. Characterization of the interface between normal and transformed epithelial cells. Nature Cell Biol. 11, 460–467 (2009).

    Article  CAS  Google Scholar 

  36. Wang, S. P. et al. p53 controls cancer cell invasion by inducing the MDM2-mediated degradation of Slug. Nature Cell Biol. 11, 694–704 (2009).

    Article  CAS  Google Scholar 

  37. Köprunner, M., Thisse, C., Thisse, B. & Raz, E. A zebrafish nanos-related gene is essential for the development of primordial germ cells. Genes Dev. 15, 2877–2885 (2001).

    PubMed  PubMed Central  Google Scholar 

  38. Nguyen, A. W. & Daugherty, P. S. Evolutionary optimization of fluorescent proteins for intracellular FRET. Nature Biotechnol. 23, 355–360 (2005).

    Article  CAS  Google Scholar 

  39. Riedl, J. et al. Lifeact: a versatile marker to visualize F-actin. Nature Methods 5, 605–607 (2008).

    Article  CAS  Google Scholar 

  40. Daoudi, M. et al. Enhanced adhesive capacities of the naturally occurring Ile249-Met280 variant of the chemokine receptor CX3CR1. J. Biol. Chem. 279, 19649–19657 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The FRET biosensors for Rac and RhoA were a gift from M. Matsuda. The yellow variant of GFP, YPet, was generously provided by P. Daugherty. Lifeact fusions were a gift from R. Wedlich-Soldner, and the GFP-CLIP-170 was provided by Franck Perez. We thank Donna Arndt-Jovin and Tom Jovin for their help with the FRET experiments. This work is supported by grants from the Deutsche Forschungsgemeinschaft (DFG) and the Max Planck Society. E.K. and B.B. are students at the International Max Planck Research School, Göttingen.

Author information

Authors and Affiliations

  1. Institute for Cell Biology, Center for Molecular Biology of Inflammation, Von-Esmarch-Strasse 56, D-48149, Münster, Germany

    Elena Kardash, Michal Reichman-Fried, Bijan Boldajipour, Esther-Maria Messerschmidt & Erez Raz

  2. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany

    Jean-Léon Maître, Ekaterina Papusheva & Carl-Philipp Heisenberg

Authors
  1. Elena Kardash
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michal Reichman-Fried
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jean-Léon Maître
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bijan Boldajipour
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ekaterina Papusheva
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Esther-Maria Messerschmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Carl-Philipp Heisenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Erez Raz
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

E.K. performed all the experiments except for the following: the initial characterization of Rac and RhoA phenotypes, generation of enhanced green fluorescent protein–actin transgenic line, track analysis in Figs 1, 2 and 4 and Supplementary Information, Fig. S7c, and in situ analysis in Supplementary Information, Fig. S1b were performed by M.R.; cell behaviour in Matrigel was conducted by B.B., adhesion measurements were performed by J.-L.M. and C.-P. H.; and the experiment described in Supplementary Information, Fig. S7a was performed by E.P. and C.-P.H. E.M. cloned DNA constructs and performed RNA injections. E.K., M.R. and E.R. wrote the manuscript.

Corresponding author

Correspondence to Erez Raz.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1722 kb)

Supplementary Information

Supplementary Movie 1 (MOV 5571 kb)

Supplementary Information

Supplementary Movie 2 (MOV 2590 kb)

Supplementary Information

Supplementary Movie 3 (MOV 2770 kb)

Supplementary Information

Supplementary Movie 4 (MOV 252 kb)

Supplementary Information

Supplementary Movie 5 (MOV 936 kb)

Supplementary Information

Supplementary Movie 6 (MOV 1101 kb)

Supplementary Information

Supplementary Movie 7 (MOV 1827 kb)

Supplementary Information

Supplementary Movie 8 (MOV 4083 kb)

Supplementary Information

Supplementary Movie 9 (MOV 7840 kb)

Supplementary Information

Supplementary Movie 10 (MOV 8964 kb)

Supplementary Information

Supplementary Movie 11 (MOV 8915 kb)

Supplementary Information

Supplementary Movie 12 (MOV 7483 kb)

Supplementary Information

Supplementary Movie 13 (MOV 7177 kb)

Supplementary Information

Supplementary Movie 14 (MOV 8484 kb)

Supplementary Information

Supplementary Movie 15 (MOV 8569 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kardash, E., Reichman-Fried, M., Maître, JL. et al. A role for Rho GTPases and cell–cell adhesion in single-cell motility in vivo. Nat Cell Biol 12, 47–53 (2010). https://doi.org/10.1038/ncb2003

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb2003

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