Morphogenetic degeneracies in the actomyosin cortex
- Max Planck Institute of Molecular Cell Biology and Genetics, Germany
- Max Planck Institute for the Physics of Complex Systems, Germany
- Flatiron Institute, United States
- Newcastle University, United Kingdom
- Wellcome Trust/Cancer Research UK Gurdon Institute, United Kingdom
- Technische Universität Dresden, Germany
- IRCCS Centro Neurolesi “Bonino Pulejo”, Italy
Figures
Figure 1
Screen strategy.
(A) Flow chart describes strategy undertaken to perform candidate RNAi screen of actin binding proteins and actomyosin regulators. Time-lapse images were then used to determine flow properties of …
Figure 2 with 3 supplements
Quantification of cortical flow velocities.
(A), (C) Comparison of mean AP velocity and mean chiral counter-rotation velocity respectively. Shaded areas in the inset represent regions over which the spatial average was performed in each …
Figure 2—figure supplement 1
Comparison of mean AP velocity, .
Each graph presents the instantaneous AP velocity histogram for the RNAi condition specified. Only ABP knockdowns that are significantly different from the negative control, klp-1 (RNAi), are …
Figure 2—figure supplement 2
Comparison of mean chiral counter-rotation velocity, .
Each graph presents the instantaneous chiral counter-rotation velocity histogram for the RNAi condition specified. Only ABP knockdowns that are significantly different from the negative control, k…
Figure 2—figure supplement 3
Comparison of mean magnitude velocity, .
Error bars, SEM; gray bar, negative control, klp-1 (RNAi) condition; gray horizontal bar, error of the mean with 99% confidence for klp-1 (RNAi); cyan, beige bars, significantly different knockdowns …
Figure 3
Quantification of pulsatile period of cortical flow.
(A) Comparison of mean period of pulsatile AP velocity, , in the posterior of the embryo. Shaded area in the inset represents the region over which the spatial average of velocity was performed in …
Figure 4 with 1 supplement
Quantification of myosin foci size.
(A) Representative 2D myosin fluorescence intensity autocorrelation function as a heat map (middle) for the anterior region (left) of a single frame during cortical flow from a klp-1 (RNAi) embryo. S…
Figure 4—figure supplement 1
Comparison of mean foci size, .
Each graph presents the foci size histogram for the RNAi condition specified. Only ABP knockdowns that are significantly different from the negative control, klp-1 (RNAi), are shown. Gray histogram, …
Figure 5 with 1 supplement
Quantification of myosin foci number density.
(A) Detection of myosin foci (right) by analysis of local changes in fluorescence intensity in the vicinity of identified local extrema (middle) for the anterior region (left) of a single frame …
Figure 5—figure supplement 1
Comparison of mean foci number density, .
Each graph presents the foci number density histogram for the RNAi condition specified. Only ABP knockdowns that are significantly different from the negative control, klp-1 (RNAi), are shown. Gray …
Figure 6 with 1 supplement
Estimation of physical properties of the cortex.
(A), (C) Comparison of the hydrodynamic length, and chirality index, of the cortex respectively. Error bars, SEM; gray bar, negative control, klp-1 (RNAi) condition; gray horizontal bar, error …
Figure 6—figure supplement 1
Comparison of hydrodynamic model fit to experimental flow profiles.
Row 1 (row 2) represents the top three ABP knockdowns that resulted in a significantly higher (lower) , and row 3 (row 4) represents the top three ABP knockdowns that resulted in a significantly …
Figure 7 with 4 supplements
Distinct molecular functions possess similar roles at large-scales.
(A) Non-linear dimensional reduction of the quantitative data set was performed and plotted along dimensions that represent the highest variability. Circular markers, ABPs; star markers, actomyosin …
Figure 7—figure supplement 1
Heat map of proteins part of the wild type cluster.
Parameters that were used for dimensional reduction (foci density , AP velocity , chirality index , flow period , hydrodynamic length ) are displayed as heat maps for knockdowns that …
Figure 7—figure supplement 2
Correlation between quantifications for ABPs.
(A) Pearson’s linear correlation coefficients between pairs of quantifications are shown as a heat map. (B)-(E) Selected pairs of quantifications that were positively or negatively correlated are …
Figure 7—figure supplement 3
Cytoplasmic actin fluorescence.
SWG007 transgenic line was used to image cytoplasmic actin. A snapshot of the indicated knockdown was performed 2 µm deeper from the cortical region at the start of cortical flows and displayed. An …
Figure 7—figure supplement 4
Correlation between quantifications and clustering dimensions.
Correlation between the five quantifications (foci density , AP velocity , chirality index , flow period , hydrodynamic length ) used for clustering and the two principal axes (D1, D2) in …
Videos
Video 1
Phenotype - Fast AP flow: Comparison of cortical flow using TH455 transgenic line (NMY-2::GFP is visualized here) between the negative control, klp-1 (RNAi) and arx-1 (RNAi) and unc-59 (RNAi), which led to significantly faster anterior-directed flow velocities.
Movies were acquired with a 5 s frame interval and are being played 75x faster than real time. Scale bar, 10 µm.
Video 2
Phenotype - Fast chiral flow: Comparison of cortical flow using TH455 transgenic line (NMY-2::GFP is visualized here) between the negative control, klp-1 (RNAi) and nex-1 (RNAi) and csnk-1 (RNAi), which led to significantly faster chiral flow velocities.
Movies were acquired with a 5 s frame interval and are being played 75x faster than real time. Scale bar, 10 µm.
Video 3
Phenotype - Slow AP and chiral flow: Comparison of cortical flow using TH455 transgenic line (NMY-2::GFP is visualized here) between the negative control, klp-1 (RNAi) and cap-1 (RNAi), unc-60 (RNAi) and ani-1 (RNAi), which led to significantly slower anterior-directed as well as chiral flow velocities.
Movies were acquired with a 5 s frame interval and are being played 75x faster than real time. Scale bar, 10 µm.
Video 4
Phenotype - Short-range flow: Comparison of cortical flow using TH455 transgenic line (NMY-2::GFP is visualized here) between the negative control, klp-1 (RNAi) and cyk-1 (RNAi), plst-1 (RNAi) and pfn-1 (RNAi), which led to short-range flow.
Movies were acquired with a 5 s frame interval and are being played 75x faster than real time. Scale bar, 10 µm.
Video 5
Pulsatile flow: Cortical flow exhibits pulsatile characteristics in the posterior cortex.
Cortical flow acquired using TH455 transgenic line (NMY-2::GFP is visualized here for the negative control, klp-1 (RNAi)) with a 5 s frame interval. Movie is being played 25x faster than real time. …
Additional files
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Supplementary file 1
Table of ABPs and suppressor proteins.
ABPs and suppressor proteins that were tested as well as those that were discarded are compiled. Common names of the proteins, their orthologous C. elegans gene names, putative functions, number of hours of RNAi and the number of biological replicates performed for each knockdown are indicated. Star indicates ABPs that were also picked up in the suppressor screen previously performed in the Ahringer lab.
- https://doi.org/10.7554/eLife.37677.024
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Transparent reporting form
- https://doi.org/10.7554/eLife.37677.025
