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