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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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.
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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.
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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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DOI: https://doi.org/10.1038/nature09151
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