Supplementary Materials

Forces Driving Epithelial Spreading in Zebrafish Gastrulation

Martin Behrndt, Guillaume Salbreux, Pedro Campinho, Robert Hauschild, Felix Oswald, Julia Roensch, Stephan W. Grill, Carl-Philipp Heisenberg

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

Download Supplement
  • Supplementary Text
  • Figs. S1 to S16
  • Materials and Methods
  • Captions for movies S1 to S10
  • References (1936)

Images, Video, and Other Other Media

Movie S1
Whole embryo confocal time-lapse movie of Tg(actb1:myl12.1-eGFP) labeling Myosin-2 throughout the course of epiboly (40-100% epiboly). Left: Lateral view of the embryo, which is slightly tilted to keep the actomyosin ring close to the objective throughout the acquisition. Right: Orthogonal view to illustrate the 3D orientation of the embryo. Scale bar, 100 µm.
Movie S2
Whole embryo confocal time-lapse movie of Tg(actb1:GFP-utrCH) labeling F-Actin throughout the course of epiboly (40-90% epiboly). Left: Lateral view of the embryo, which is slightly tilted to keep the actomyosin ring close to the objective throughout the acquisition. Right: Orthogonal view to illustrate the 3D orientation of the embryo. Scale bar, 100 µm.
Movie S3
Local disruption of the actomyosin ring by consecutive UV laser ablations in a Tg(actb1:myl12.1-eGFP) embryo labeling Myosin-2 at 60% epiboly. Scale bar, 50 µm; white rectangle indicates onset of ablation.
Movie S4
Cortical laser ablation of a Tg(actb1:myl12.1-eGFP) embryo labeling Myosin-2 at 60-70% epiboly to assess circumferential tension within the actomyosin ring. Red line (20 µm) indicates where the cut will be performed. Scale bar, 10 µm.
Movie S5
Cortical laser ablation of a Tg(actb1:myl12.1-eGFP) embryo labeling Myosin-2 at 60-70% epiboly to assess AV tension within the actomyosin ring. Green line (20 µm) indicates where the cut will be performed. Scale bar, 10 µm.
Movie S6
Cortical laser ablation to assess circumferential tension within the actomyosin ring of a Tg(actb1:myl12.1-mCherry) embryo labeling Myosin-2 at 60-70% epiboly, which was treated with the myosin 49 inhibitor Blebbistatin (-) (20 μM). Red line (20 µm) indicates where the cut will be performed. Scale bar, 10 µm.
Movie S7
Cortical laser ablation to assess AV tension within the actomyosin ring of a Tg(actb1:myl12.1-­mCherry) embryo labeling Myosin-2, which was treated with the myosin inhibitor Blebbistatin (-) (20 µM). Green line (20 µm) indicates where the cut will be performed. Scale bar, 10 µm.
Movie S8
High-resolution spinning disk time-lapse of a Tg(actb1:myl12.1-eGFP) labeling Myosin-2 (left) injected with lifeact-RFP mRNA labeling F-Actin (middle) at 60-70% epiboly. In the merged dual-color movie (right) Myosin-2 is depicted in green and F-Actin in red. White rectangle indicates the region for the magnified view (bottom on the respective movies; for better visualization brightness was dynamically adjusted for the magnified view only). Bright F-actin patches moving with the F-actin subnetwork may represent endocytic pits. Note, that the frame rate is discontinuous as image acquisition was performed by looping through multiple positions (see fig. S8). Scale bars, 10 µm.
Movie S9
Cortical laser ablation of a Tg(actb1:myl12.1-eGFP) embryo labeling Myosin-2 at 75% epiboly reveal vanishing tension at the vegetal pole. White line (20 µm) indicates where the cut will be performed. Scale bar, 10 µm.
Movie S10
High-resolution spinning disk time-lapse of a cylindrically shaped Tg(actb1:GFP-utrCH) embryo labeling F-Actin at 60-70% epiboly (left). White rectangle indicates the magnified view (right) on the EVL margin. Note, that the frame rate is discontinuous as image acquisition was performed by looping through multiple positions (see fig. S8). Scale bars, 20 µm.