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Fgf8 morphogen gradient forms by a source-sink mechanism with freely diffusing molecules

Nature volume 461, pages 533–536 (2009)Cite this article

Abstract

It is widely accepted that tissue differentiation and morphogenesis in multicellular organisms are regulated by tightly controlled concentration gradients of morphogens1,2. How exactly these gradients are formed, however, remains unclear3,4,5,6,7,8,9,10,11,12. Here we show that Fgf8 morphogen gradients in living zebrafish embryos are established and maintained by two essential factors: fast, free diffusion of single molecules away from the source through extracellular space, and a sink function of the receiving cells, regulated by receptor-mediated endocytosis. Evidence is provided by directly examining single molecules of Fgf8 in living tissue by fluorescence correlation spectroscopy, quantifying their local mobility and concentration with high precision. By changing the degree of uptake of Fgf8 into its target cells, we are able to alter the shape of the Fgf8 gradient. Our results demonstrate that a freely diffusing morphogen can set up concentration gradients in a complex multicellular tissue by a simple source-sink mechanism.

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Figure 1: FCS setup to investigate morphogen characteristics.
Figure 2: Labelling strategy for Fgf8.
Figure 3: Fgf8–EGFP diffuse as single molecules in the extracellular space.
Figure 4: Endocytosis regulates extracellular Fgf8 concentration gradient.

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Acknowledgements

We thank members of the Brand and Schwille laboratories for discussions; A. Picker and K. Heinze for help with initial FCS measurements; B. Lutz for discussions; A. Oates, C. Boekel, A. Picker and P. Scott for comments on the manuscript; M. Fischer and K. Sipple for fish care; and A. Machate and B. Borgonov for technical assistance. This work was supported by an HFSP network grant (050503-50) to M.Br. and P.S., and by the EU Endotrack grant (050503-52) to M.Br.

Author Contributions S.R.Y. and M.Bu. performed the experiments, analysed the data and wrote the paper. M.N., J.R. and Z.P. provided advice on experimental design, data evaluation and interpretation. S.S. provided protocols for initial FCS measurements. P.S. and M.Br. initiated and supervised the collaboration, designed the project, analysed the data and wrote the paper.

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Author notes

  1. Steffen Scholpp

    Present address: Present address: Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe, D-76021 Karlsruhe, Germany.,

  2. Shuizi Rachel Yu and Markus Burkhardt: These authors contributed equally to this work.

Authors and Affiliations

  1. Developmental Genetics, Biotechnology Center, TUD,,

    Shuizi Rachel Yu, Matthias Nowak, Steffen Scholpp & Michael Brand

  2. Biophysics, Biotechnology Center, TUD, and,,

    Markus Burkhardt, Jonas Ries, Zdeněk Petrášek & Petra Schwille

  3. Center for Regenerative Therapies, TUD, Tatzberg 47-49, 01307 Dresden, Germany ,

    Shuizi Rachel Yu, Markus Burkhardt, Matthias Nowak, Jonas Ries, Zdeněk Petrášek, Petra Schwille & Michael Brand

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  1. Shuizi Rachel Yu
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Correspondence to Petra Schwille or Michael Brand.

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This file contains Supplementary Notes 1-8, Supplementary References and Supplementary Figures 1-10 with Legends. (PDF 1094 kb)

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Yu, S., Burkhardt, M., Nowak, M. et al. Fgf8 morphogen gradient forms by a source-sink mechanism with freely diffusing molecules. Nature 461, 533–536 (2009). https://doi.org/10.1038/nature08391

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