Letter | Published:

Planar polarized actomyosin contractile flows control epithelial junction remodelling

Nature volume 468, pages 11101114 (23 December 2010) | Download Citation

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Abstract

Force generation by Myosin-II motors on actin filaments drives cell and tissue morphogenesis1,2,3,4,5,6,7,8,9,10,11,12,13,14,15. In epithelia, contractile forces are resisted at apical junctions by adhesive forces dependent on E-cadherin16, which also transmits tension6,17,18,19. During Drosophila embryonic germband extension, tissue elongation is driven by cell intercalation20, which requires an irreversible and planar polarized remodelling of epithelial cell junctions4,5. We investigate how cell deformations emerge from the interplay between force generation and cortical force transmission during this remodelling in Drosophila melanogaster. The shrinkage of dorsal–ventral-oriented (‘vertical’) junctions during this process is known to require planar polarized junctional contractility by Myosin II (refs 4, 5, 7, 12). Here we show that this shrinkage is not produced by junctional Myosin II itself, but by the polarized flow of medial actomyosin pulses towards ‘vertical’ junctions. This anisotropic flow is oriented by the planar polarized distribution of E-cadherin complexes, in that medial Myosin II flows towards ‘vertical’ junctions, which have relatively less E-cadherin than transverse junctions. Our evidence suggests that the medial flow pattern reflects equilibrium properties of force transmission and coupling to E-cadherin by α-Catenin. Thus, epithelial morphogenesis is not properly reflected by Myosin II steady state distribution but by polarized contractile actomyosin flows that emerge from interactions between E-cadherin and actomyosin networks.

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Change history

  • 17 February 2011

    The present address for author M.Z. was corrected on 17 February 2011

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Acknowledgements

We thank R. Levayer, M. Mavrakis and J.-M. Philippe for making and sharing respectively the utrophin–GFP, GAP43–Venus and GAP43–Cherry transgenic lines. We are grateful to our colleagues for their gifts of reagents: B. Bement, R. Karess, H. Oda, A. Martin and E. Wieschaus. We thank E. Munro and all members of the Lecuit and Lenne groups for discussions, and comments on the manuscript. This work was supported by a Programme Grant from HFSP to T.L, P.-F.L and E. Munro, by the CNRS, the Fondation pour la Recherche Médicale (to T.L), the ANR-Blanc 2005 (to T.L. and P.-F.L), Région PACA and ANR-PCV 2008 (to P.-F.L and T.L.). M.R. was supported by a PhD fellowship by the Région PACA and Amplitude Systems.

Author information

Author notes

    • Matteo Rauzi

    Present address: EMBL, Meyerhofstrasse 1, 69117 Heidelberg, Germany.

Affiliations

  1. IBDML, UMR6216 CNRS-Université de la Méditerranée, Campus de Luminy, case 907, 13288 Marseille Cedex 09, France

    • Matteo Rauzi
    • , Pierre-François Lenne
    •  & Thomas Lecuit

Authors

  1. Search for Matteo Rauzi in:

  2. Search for Pierre-François Lenne in:

  3. Search for Thomas Lecuit in:

Contributions

M.R. made the original observations of pulsed contractility and flow. M.R. and T.L. planned the project, and analysed the data together with P.-F.L.; M.R. conducted the experiments except for FRAP experiments on MRLC–GFP, which were performed by P.-F.L. and T.L.; P.-F.L. and M.R. developed the nano-ablation system. T.L. and P.-F.L. wrote the manuscript together with M.R. All authors commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Pierre-François Lenne or Thomas Lecuit.

Supplementary information

PDF files

  1. 1.

    Supplementary Figures

    The file contains Supplementary Figures 1-12 with legends.

Videos

  1. 1.

    Supplementary Movie 1a

    Time lapse movie of Utrophin::GFP (labelling F-actin) in intercalating cells of a wild type embryo, 15-20 min after the onset of gastrulation. The focal plane is at the apical most position near the plasma membrane. Scale bar: 5 μm.

  2. 2.

    Supplementary Movie 1b

    Time lapse movie of Utrophin::GFP (labelling F-actin) in intercalating cells of a wild type embryo, 15-20 min after the onset of gastrulation. The focal plane is at a slightly less apical focal plane, intersecting the adherens junctions. Scale bar: 5 μm.

  3. 3.

    Supplementary Movie 2

    Time lapse movie of MRLC::GFP (Myo-II) in intercalating cells of a wild type embryo, 15-20 min after the onset of gastrulation. Each image is the projection of a 3 μm zstack. Scale bar: 5 μm.

  4. 4.

    Supplementary Movie 3

    Time lapse movie of MRLC::Cherry (red) and Utr ::GFP (green) in the apical region of intercalating cells of a wild type embryo

  5. 5.

    Supplementary Movie 4

    Time lapse movie of MRLC::GFP (Myo-II) in intercalating cells of a wild type embryo. Nano dissection (< 400 nm, red arrowhead) of medial Myo-II, 15-20 min after the onset of gastrulation. Scale bar: 5μm.

  6. 6.

    Supplementary Movie 5

    Time lapse movie of MRLC::GFP (Myo-II) in intercalating cells of a wild type embryo,, 20 min after the onset of gastrulation. Each image is the projection of a 3 μm zstack.. Coloured traces represent the trajectory of coalescing speckles of medial Myo- II. Scale bar: 5 μm.

  7. 7.

    Supplementary Movie 6

    Time lapse movie of MRLC::GFP (Myo-II) in cells of a krüppel RNAi injected embryo,15-20 min after the onset of gastrulation. Each image is the projection of a 3 μm z-stack. Scale bar: 5 μm.

  8. 8.

    Supplementary Movie 7

    Time lapse movie of MRLC::GFP (Myo-II) in cells of a wild type embryo. Each image is the projection of a 3 μm z-stack. The most apical planes of the 3 μm stack (500 nm projection) are shown in green while the most basal (2.5 μm projection) are shown in red. Scale bar: 5 μm.

  9. 9.

    Supplementary Movie 8

    Time lapse movie and FRAP of MRLC::GFP (Myo-II) in cells of a wild type embryo. Each image is the projection of a 3 μm z-stack. FRAP is performed on the junctional pool of Myo-II at time 0 seconds. Clusters of Myo-II from the medial pool are then transferred to the cortex. Scale bar: 5 μm.

  10. 10.

    Supplementary Movie 9

    Time lapse movie of GAP43::Venus (plasma membrane) in the apical region of intercalating cells. Note the relative flat organization of the apical surface. Scale bar: 5 μm.

  11. 11.

    Supplementary Movie 10

    Time lapse movie of GAP43::Venus (plasma membrane) in the apical region of apically constricting mesoderm cells. Note the strongly ruffled apical surface. Scale bar: 5μm.

  12. 12.

    Supplementary Movie 11

    Time lapse movie of GAP43::Cherry (plasma membrane, red, middle) and Utr::GFP (F-actin, green, right) in the apical region of intercalating cells. The trajectories of a small protrusion (red dot in middle panel) and of an actin patch (green dot in right panel) are shown. Scale bar: 5 μm.

  13. 13.

    Supplementary Movie 12

    Time lapse movie of GAP43::Cherry (plasma membrane, red, middle) and Utr::GFP (F-actin, green, right) in the apical region of intercalating cells. The trajectories of a small protrusion (red dot in middle panel) and of an actin patch (green dot in right panel) are shown. Scale bar: 5 μm.

  14. 14.

    Supplementary Movie 13

    Time lapse movie of MRLC::Cherry (Myo-II) and E-cad::GFP in intercalating cells of a wild type embryo, 15-20 min after the onset of gastrulation. Each image is the projection of a 3 μm z-stack. Left panel: merged signal of MRLC::Cherry (green) and E-cad::GFP (red); central panel: E-cad::GFP; right panel: MRLC::Cherry. Scale bar: 5 μm.

  15. 15.

    Supplementary Movie 14

    Time lapse movie of MRLC::Cherry (displayed in green) and E-cad::GFP (displayed in red) in an E-cad RNAi embryo, 15-20 min after the presumptive onset of gastrulation. Each image is the projection of a 3 μm z-stack. Scale bar: 5 μm.

  16. 16.

    Supplementary Movie 15

    Time lapse movie of MRLC::Cherry (displayed in green) and E-cad::GFP (displayed in red) in an α-cat RNAi embryo, 15-20 min after the presumptive onset of gastrulation. Each image is the projection of a 3 μm z-stack. Scale bar: 5 μm.

  17. 17.

    Supplementary Movie 16

    Time lapse movie of MRLC::Cherry (displayed in green) and E-cad::GFP (displayed in red) in intercalating cells of a wild type embryo, 15-20 min after the onset of gastrulation. Nanodissectionof medial Myo-II is indicated by a white arrowhead. The left panel shows the trajectory of the remaining cluster of medial Myo-II after ablation. Scale bar: 5 μm.

  18. 18.

    Supplementary Movie 17

    Time lapse movie of Utr::GFP (F-actin) in intercalating cells of a wild type embryo. Nanodissection of the medial F-actin meshwork is indicated by a red arrowhead. Note the relaxation of the meshwork following ablation. Scale bar: 5 μm.

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DOI

https://doi.org/10.1038/nature09566

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