Letter | Published:

Attachment of the blastoderm to the vitelline envelope affects gastrulation of insects

Naturevolume 568pages395399 (2019) | Download Citation

Abstract

During gastrulation, physical forces reshape the simple embryonic tissue to form the complex body plans of multicellular organisms1. These forces often cause large-scale asymmetric movements of the embryonic tissue2,3. In many embryos, the gastrulating tissue is surrounded by a rigid protective shell4. Although it is well-recognized that gastrulation movements depend on forces that are generated by tissue-intrinsic contractility5,6, it is not known whether interactions between the tissue and the protective shell provide additional forces that affect gastrulation. Here we show that a particular part of the blastoderm tissue of the red flour beetle (Tribolium castaneum) tightly adheres in a temporally coordinated manner to the vitelline envelope that surrounds the embryo. This attachment generates an additional force that counteracts tissue-intrinsic contractile forces to create asymmetric tissue movements. This localized attachment depends on an αPS2 integrin (inflated), and the knockdown of this integrin leads to a gastrulation phenotype that is consistent with complete loss of attachment. Furthermore, analysis of another integrin (the αPS3 integrin, scab) in the fruit fly (Drosophila melanogaster) suggests that gastrulation in this organism also relies on adhesion between the blastoderm and the vitelline envelope. Our findings reveal a conserved mechanism through which the spatiotemporal pattern of tissue adhesion to the vitelline envelope provides controllable, counteracting forces that shape gastrulation movements in insects.

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Data availability

All raw imaging data are available from P.T. upon request.

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Change history

  • 10 April 2019

    In this Letter, the sentence starting: ‘For instance, Tribolium and Drosophila inflated are direct targets of the mesoderm…’ has been corrected online; see accompanying Amendment.

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Acknowledgements

We thank M. van der Zee for the transgenic LifeAct-eGFP Tribolium line; Y. Hsieh for mRNA, P. Mejstrik, T. Pietzsch, M. Burkon and the MPI-CBG Electron and Light microscopy facilities for technical assistance; M. Benton, K. Panfilio, L. Jawerth and P. Gross for helpful discussions; the Tribolium research community for support; and C. Norden, E. Knust and C. Zechner for comments on the manuscript. A.J. received a DIGS-BB fellowship, and S.M. was supported by an ELBE post-doctoral fellowship. S.W.G. acknowledges support from the European Research Council (CHIMO, grant No 742712) and the Deutsche Forschungsgemeinschaft (DFG) under Germany´s Excellence Strategy – EXC-2068 – 390729961.

Reviewer information

Nature thanks Kristen Panfilio, Siegfried Roth and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Author information

Author notes

  1. These authors contributed equally: Akanksha Jain, Alexander Mietke

Affiliations

  1. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    • Stefan Münster
    • , Akanksha Jain
    • , Alexander Mietke
    • , Stephan W. Grill
    •  & Pavel Tomancak
  2. Max Planck Institute for the Physics of Complex Systems, Dresden, Germany

    • Stefan Münster
    •  & Alexander Mietke
  3. Center for Systems Biology, Dresden, Germany

    • Stefan Münster
    • , Alexander Mietke
    • , Stephan W. Grill
    •  & Pavel Tomancak
  4. Biotechnology Center, TU Dresden, Dresden, Germany

    • Stefan Münster
    •  & Stephan W. Grill
  5. Faculty of Scientific Computing, TU Dresden, Dresden, Germany

    • Alexander Mietke
  6. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA

    • Anastasios Pavlopoulos
  7. Cluster of Excellence Physics of Life, TU Dresden, Dresden, Germany

    • Stephan W. Grill

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Contributions

S.M. designed the research, performed experiments, analysed the data and wrote the manuscript. A.J. produced reagents and performed experiments. A.M. developed the theory and analysed the data. A.P. suggested the project and produced reagents. S.W.G. and P.T. conceived and oversaw the project, designed the research and wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Stephan W. Grill or Pavel Tomancak.

Extended data figures and tables

  1. Extended Data Fig. 1 Imaging results and theoretical modelling for several Tribolium wild-type specimens.

    ac, Kymographs of myosin intensity (coloured according to the colour bar at the bottom) along the contour of the blastoderm for three different Tribolium wild-type specimens injected with mRNA that encodes for Tcsqh-eGFP, recorded with light-sheet microscopy at 25 °C. White lines show flow, measured by particle image velocimetry tracking. The horizontal dashed lines denote the individual time points displayed in df. Panels ac represent three independent experiments, n = 3. df, Experimentally determined myosin intensity (colour) as well as tissue flow field (arrows) for single time points of ac. Insets, experimentally determined flow field compared to theoretical prediction, assuming the tissue is anchored at the anterior–ventral side of the egg (red anchor). Fitting parameters were vc = 0.35 μm s−1, α < 0.2 (d); vc = 0.88 μm s−1, α < 0.2 (e); and vc = 0.45 μm s−1, α < 0.2 (f). Data from a and d are shown in Fig. 1c, e, g and Supplementary Video 3. Data from b are shown in Supplementary Video 1. Note that our theory can recapitulate the flow fields of individual embryos, as opposed to using ensemble-averaged data.

  2. Extended Data Fig. 2 Imaging results and theoretical modelling for several Tribolium specimens with Tcif knockdown.

    ac, Kymographs of myosin intensity (coloured according to the colour bar at the bottom) along the contour of the blastoderm for three different Tribolium specimens injected with a mixture of mRNA that encodes for Tcsqh-eGFP and iBeetle RNAi against Tcif, recorded with light-sheet microscopy. The temperature of experiments was 30 °C (a) or 25 °C (b, c). White lines show flow, measured by particle image velocimetry tracking. The horizontal dashed lines denote the individual time points displayed in df. Panels ac represent three independent experiments, n = 3. df, Experimentally determined myosin intensity (colour) as well as tissue flow field (arrows) for single time points of ac. Insets, experimentally determined flow field compared to theoretical prediction assuming the tissue is free to flow with respect to the vitelline envelope. Fitting parameters were vc = 0.45 μm s−1, α < 0.2 (d); vc = 0.58 μm s−1, α < 0.2 (e); vc = 0.39 μm s−1, α < 0.2 (f). Data from d are shown in Fig. 3g and Supplementary Video 12.

Supplementary information

  1. Supplementary Information

    This file contains Supplementary Methods: Active fluid theory for Tribolium tissue flow.

  2. Reporting Summary

  3. Supplementary Information

    This file contains Supplementary Video Captions for Supplementary Videos 1-16.

  4. Video 1

    Dimensionality reduction for theory.

  5. Video 2

    Comparison of experiment and theory without attachment.

  6. Video 3

    Comparison of experiment and theory with attachment.

  7. Video 4

    Dynamics of apical protrusions in Tribolium blastoderm.

  8. Video 5

    Dynamics of blastoderm vitelline attachment in Tribolium.

  9. Video 6

    Blastoderm vitelline proximity map in Tribolium.

  10. Video 7

    Attachment rip-off during serosa window closure in Tribolium.

  11. Video 8

    Cross-section view of attachment rip-off during serosa window closure in Tribolium.

  12. Video 9

    Attachment disruption by trypsin digestion in Tribolium.

  13. Video 10

    Release of attachment by Tcif RNAi in Tribolium I.

  14. Video 11

    Release of attachment by Tcif RNAi in Tribolium II.

  15. Video 12

    Comparison of experiment and theory in Tcif knockdown embryos.

  16. Video 13

    Blastoderm vitelline proximity map in Drosophila.

  17. Video 14

    Trypsin injection into perivitelline space in Drosophila.

  18. Video 15

    Twisted gastrulation phenotype in Drosophila embryos mutant for scab I.

  19. Video 16

    Twisted gastrulation phenotype in Drosophila embryos mutant for scab II.

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https://doi.org/10.1038/s41586-019-1044-3

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