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A random cell motility gradient downstream of FGF controls elongation of an amniote embryo

Nature volume 466, pages 248–252 (2010)Cite this article

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Abstract

Vertebrate embryos are characterized by an elongated antero-posterior (AP) body axis, which forms by progressive cell deposition from a posterior growth zone in the embryo. Here, we used tissue ablation in the chicken embryo to demonstrate that the caudal presomitic mesoderm (PSM) has a key role in axis elongation. Using time-lapse microscopy, we analysed the movements of fluorescently labelled cells in the PSM during embryo elongation, which revealed a clear posterior-to-anterior gradient of cell motility and directionality in the PSM. We tracked the movement of the PSM extracellular matrix in parallel with the labelled cells and subtracted the extracellular matrix movement from the global motion of cells. After subtraction, cell motility remained graded but lacked directionality, indicating that the posterior cell movements associated with axis elongation in the PSM are not intrinsic but reflect tissue deformation. The gradient of cell motion along the PSM parallels the fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK) gradient1, which has been implicated in the control of cell motility in this tissue2. Both FGF signalling gain- and loss-of-function experiments lead to disruption of the motility gradient and a slowing down of axis elongation. Furthermore, embryos treated with cell movement inhibitors (blebbistatin or RhoK inhibitor), but not cell cycle inhibitors, show a slower axis elongation rate. We propose that the gradient of random cell motility downstream of FGF signalling in the PSM controls posterior elongation in the amniote embryo. Our data indicate that tissue elongation is an emergent property that arises from the collective regulation of graded, random cell motion rather than by the regulation of directionality of individual cellular movements.

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Figure 1: Role of caudal PSM in embryo elongation.
Figure 2: Posterior-to-anterior decreasing gradient of random motility in the PSM.
Figure 3: Effect of FGF signalling, cell movement and cell proliferation on axis elongation.
Figure 4: Simulation results at t = 0,50,100,250.

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References

  1. Dubrulle, J. & Pourquie, O. fgf8 mRNA decay establishes a gradient that couples axial elongation to patterning in the vertebrate embryo. Nature 427, 419–422 (2004)

    Article  ADS  CAS  Google Scholar 

  2. Delfini, M. C., Dubrulle, J., Malapert, P., Chal, J. & Pourquie, O. Control of the segmentation process by graded MAPK/ERK activation in the chick embryo. Proc. Natl Acad. Sci. USA 102, 11343–11348 (2005)

    Article  ADS  CAS  Google Scholar 

  3. Keller, R., Shook, D. & Skoglund, P. The forces that shape embryos: physical aspects of convergent extension by cell intercalation. Phys. Biol. 5, 015007 (2008)

    Article  ADS  Google Scholar 

  4. Lawson, A. & Schoenwolf, G. C. New insights into critical events of avian gastrulation. Anat. Rec. 262, 238–252 (2001)

    Article  CAS  Google Scholar 

  5. Voiculescu, O., Bertocchini, F., Wolpert, L., Keller, R. E. & Stern, C. D. The amniote primitive streak is defined by epithelial cell intercalation before gastrulation. Nature 449, 1049–1052 (2007)

    Article  ADS  CAS  Google Scholar 

  6. Yang, X., Dormann, D., Munsterberg, A. E. & Weijer, C. J. Cell movement patterns during gastrulation in the chick are controlled by positive and negative chemotaxis mediated by FGF4 and FGF8. Dev. Cell 3, 425–437 (2002)

    Article  CAS  Google Scholar 

  7. Beddington, R. S. Induction of a second neural axis by the mouse node. Development 120, 613–620 (1994)

    CAS  PubMed  Google Scholar 

  8. Waddington, C. H. Experiments on the development of chick and duck embryos cultivated in vitro. Philos. Trans. R. Soc. Lond. B 221, 179–230 (1932)

    Article  ADS  Google Scholar 

  9. Charrier, J. B., Teillet, M. A., Lapointe, F. & Le Douarin, N. M. Defining subregions of Hensen’s node essential for caudalward movement, midline development and cell survival. Development 126, 4771–4783 (1999)

    CAS  PubMed  Google Scholar 

  10. Bellairs, R. The development of somites in the chick embryo. J. Embryol. Exp. Morphol. 11, 697–714 (1963)

    CAS  PubMed  Google Scholar 

  11. Spratt, N. T. Jr. Analysis of the organizer center in the early chick embryo. I. Localisation of prospective notochord and somite cells. J. Exp. Zool. 128, 121–163 (1955)

    Article  Google Scholar 

  12. Hamburger, V. The stage series of the chick embryo. Dev. Dyn. 195, 273–275 (1992)

    Article  CAS  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  14. Sbalzarini, I. F. & Koumoutsakos, P. Feature point tracking and trajectory analysis for video imaging in cell biology. J. Struct. Biol. 151, 182–195 (2005)

    Article  CAS  Google Scholar 

  15. Kulesa, P. M. & Fraser, S. E. Cell dynamics during somite boundary formation revealed by time-lapse analysis. Science 298, 991–995 (2002)

    Article  ADS  CAS  Google Scholar 

  16. Rifes, P. et al. Redefining the role of ectoderm in somitogenesis: a player in the formation of the fibronectin matrix of presomitic mesoderm. Development 134, 3155–3165 (2007)

    Article  CAS  Google Scholar 

  17. Zamir, E. A., Rongish, B. J. & Little, C. D. The ECM moves during primitive streak formation–computation of ECM versus cellular motion. PLoS Biol. 6, e247 (2008)

    Article  Google Scholar 

  18. Zamir, E. A., Czirók, A., Cui, C., Little, C. D. & Rongish, B. J. Mesodermal cell displacements during avian gastrulation are due to both individual cell-autonomous and convective tissue movements. Proc. Natl Acad. Sci. USA 103, 19806–19811 (2006)

    Article  ADS  CAS  Google Scholar 

  19. Berg, H. C. Random Walks in Biology. (Princeton Univ. Press, 1993)

    Google Scholar 

  20. Czirók, A., Zamir, E. A., Filla, M. B., Little, C. D. & Rongish, B. J. Extracellular matrix macroassembly dynamics in early vertebrate embryos. Curr. Top. Dev. Biol. 73, 237–258 (2006)

    Article  Google Scholar 

  21. Amaya, E., Musci, T. J. & Kirschner, M. W. Expression of a dominant negative mutant of the FGF receptor disrupts mesoderm formation in Xenopus embryos. Cell 66, 257–270 (1991)

    Article  CAS  Google Scholar 

  22. Ciruna, B. & Rossant, J. FGF signaling regulates mesoderm cell fate specification and morphogenetic movement at the primitive streak. Dev. Cell 1, 37–49 (2001)

    Article  CAS  Google Scholar 

  23. Sun, X., Meyers, E. N., Lewandoski, M. & Martin, G. R. Targeted disruption of Fgf8 causes failure of cell migration in the gastrulating mouse embryo. Genes Dev. 13, 1834–1846 (1999)

    Article  CAS  Google Scholar 

  24. Watanabe, T. et al. Tet-on inducible system combined with in ovo electroporation dissects multiple roles of genes in somitogenesis of chicken embryos. Dev. Biol. 305, 625–636 (2007)

    Article  CAS  Google Scholar 

  25. Cai, A. Q., Landman, K. A. & Hughes, B. D. Modelling directional guidance and motility regulation in cell migration. Bull. Math. Biol. 68, 25–52 (2006)

    Article  MathSciNet  CAS  Google Scholar 

  26. Gillespie, D. T. Exact stochastic simulation of coupled chemical reactions. J. Phys. Chem. 81, 2340–2361 (1977)

    Article  CAS  Google Scholar 

  27. Chapman, S. C., Collignon, J., Schoenwolf, G. C. & Lumsden, A. Improved method for chick whole-embryo culture using a filter paper carrier. Dev. Dyn. 220, 284–289 (2001)

    Article  CAS  Google Scholar 

  28. Czirók, A., Rupp, P. A., Rongish, B. J. & Little, C. D. Multi-field 3D scanning light microscopy of early embryogenesis. J. Microsc. 206, 209–217 (2002)

    Article  MathSciNet  Google Scholar 

  29. Rupp, P. A., Rongish, B. J., Czirók, A. & Little, C. D. Culturing of avian embryos for time-lapse imaging. Biotechniques 34, 274–278 (2003)

    Article  CAS  Google Scholar 

  30. Araki, I. & Nakamura, H. Engrailed defines the position of dorsal di-mesencephalic boundary by repressing diencephalic fate. Development 126, 5127–5135 (1999)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank R. Li, D. Roellig and members of the Pourquié laboratory for critical reading of the manuscript. The authors acknowledge E. Siggia for support, B. Rongish, M. Filla, C. Cui, A. Peterson, K. Perko, R. Alexander, R. Fender, J. D. Fauny, T. Cheuvront, and members of the IGBMC imaging center for assistance with imaging-related techniques. We thank L. Kennedy and T. Iimura for help with the experiments, and the members of the Cytometry Facility at the Stowers Institute for Medical Research for assistance. We thank S. Leblanc, H. Li, N. Wicker and P. Moncuquet for help with statistical analysis. The authors thank S. Esteban for artwork and J. Chatfield for editorial assistance. This research was supported by the Howard Hughes Medical Institute, the Stowers Institute for Medical Research, NIH grant R02 HD043158 and a Chaire d’excellence to O.P.; P.F. is supported by NSF grant DMR-0129848 (attributed to E. Siggia) and Lavoisier Fellowship. R.E.B. is supported by an RCUK Fellowship in Mathematical Biology and a Microsoft European Postdoctoral Research Fellowship. C.D.L. is supported by the Mathers Charitable Foundation.

Author information

Authors and Affiliations

  1. Howard Hughes Medical Institute, Kansas City, 64110, Missouri, USA

    Bertrand Bénazéraf & Olivier Pourquié

  2. Stowers Institute for Medical Research, Kansas City, 64110, Missouri, USA

    Bertrand Bénazéraf, Nicolas Denans & Olivier Pourquié

  3. Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS (UMR 7104), Inserm U964, Université de Strasbourg, Illkirch, F-67400, France

    Bertrand Bénazéraf, Nicolas Denans & Olivier Pourquié

  4. Center for Studies in Physics and Biology, The Rockefeller University, New York, 10065, New York, USA

    Paul Francois

  5. Centre for Mathematical Biology, Mathematical Institute, University of Oxford, 24–29 St Giles’, Oxford, OX1 3LB, UK

    Ruth E. Baker

  6. Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, 66160, Kansas, USA

    Charles D. Little & Olivier Pourquié

Authors
  1. Bertrand Bénazéraf
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Contributions

B.B. and O.P. designed the biological experiments. B.B. and N.D. performed the biological experiments. C.D.L. gave technical and theoretical support for the time-lapse experiments. P.F., N.D., B.B. and O.P. analysed the biological experiments; in particular, P.F. designed the tracking programs, tracked cells and ECM motions, and performed the random walk analysis. R.E.B. designed, analysed and simulated the mathematical models in collaboration with B.B., P.F. and O.P.; B.B., O.P., R.E.B. and P.F. wrote the manuscript. All authors discussed the results and commented on the manuscript. O.P. supervised the project.

Corresponding author

Correspondence to Olivier Pourquié.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-10 with legends, Supplementary Methods and References, Supplementary Notes, which include, numerical simulation, results, data processing, figures 1-2 with legends and statistics, and full legends for Supplementary Movies 1-17. (PDF 3045 kb)

Supplementary Movie 1

Brighfield recording showing the elongation of an embryo in which the caudal PSM was ablated. (MOV 3110 kb)

Supplementary Movie 2

Brighfield recording showing the elongation of an embryo in which the axial caudal progenitor zone was ablated. (MOV 4266 kb)

Supplementary Movie 3

Brighfield recording showing the elongation of an embryo in which the lateral plates were ablated. (MOV 4980 kb)

Supplementary Movie 4

Brighfield recording showing the elongation of embryo in which the rostral PSM progenitor zone was ablated. (MOV 3880 kb)

Supplementary Movie 5

This movie shows cellular movements in the PSM during embryo elongation. (MOV 5357 kb)

Supplementary Movie 6

This movie shows cellular movements and ECM movement in the PSM during embryo elongation. (MOV 5378 kb)

Supplementary Movie 7

This movie shows fibronectin and Fibrillin-2 movement in the PSM during embryo elongation. (MOV 5593 kb)

Supplementary Movie 8

This movie shows the effect of FGFR1dn electroporation on cellular movements and embryo elongation. (MOV 4357 kb)

Supplementary Movie 9

This movie shows the effect of FGF8 electroporation on cellular movements and embryo elongation. (MOV 4357 kb)

Supplementary Movie 10

This movie shows the effect of the Rho-Kinase inhibitor Y27632 on cellular movements and embryo elongation. (MOV 4594 kb)

Supplementary Movie 11

This movie shows effect of Blebbistatin on cellular movements and embryo elongation. (MOV 6880 kb)

Supplementary Movie 12

This movie shows the effect of Mitomycin C on cellular movements and embryo elongation. (MOV 5801 kb)

Supplementary Movie 13

This movie shows the effect of Aphidicolin on cellular movements and embryo elongation. (MOV 6347 kb)

Supplementary Movie 14

This movie file contains an Illustration of directional diffusion. (MOV 4406 kb)

Supplementary Movie 15

This movie shows the mathematical simulation of PSM cellular movements with gradient of cellular motility. (MOV 545 kb)

Supplementary Movie 16

This movie shows the mathematical simulation of PSM cellular movements without gradient of cellular motility. (MOV 555 kb)

Supplementary Movie 17

This movie shows the cellular movements and tissue movement in the PSM during embryo elongation growing on New culture. (MOV 3355 kb)

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Bénazéraf, B., Francois, P., Baker, R. et al. A random cell motility gradient downstream of FGF controls elongation of an amniote embryo. Nature 466, 248–252 (2010). https://doi.org/10.1038/nature09151

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