Fig. 3: 5HT2A amplifies Toll/Cirl polarity cue. | Nature Communications

Fig. 3: 5HT2A amplifies Toll/Cirl polarity cue.

From: Serotonin signaling regulates actomyosin contractility during morphogenesis in evolutionarily divergent lineages

Fig. 3

a Schematic showing Toll/Cirl mutual attraction recruiting MyoII at the cell-cell interface. b–d 5HT2A overexpression and Toll-2,6,8 knockdown. b MyoII images in different conditions (from left to right respectively): water injected WT, water injected 5HT2A + +, Toll 2,6,8 RNAi injected WT, and Toll 2,6,8 RNAi injected 5HT2A + + embryos. Quantification of MyoII intensities in DV and AP oriented junctions over-time (c), amplitude of polarity (d) in different conditions. e, f Genetic interaction between Cirl and 5HT2A. e MyoII snapshots in different conditions (from left to right respectively): control, Cirl−/−, 5HT2A−/−, Cirl−/+ 5HT2A−/+, Cirl-/+ 5HT2A−/− and Cirl−/− 5HT2A−/+. f Quantification of junctional MyoII intensities in different genotypes. g Quantification of local tissue extension. Representative images are average projection of Sqh::mCherry. The centroid of the cells outlined in dark was tracked every 5 min for 15 min starting 30 min after the cellularization front passed the nucleus, pink dotted lines are the track of the centroids. The relative length (L-L0)/L0 is plotted at the bottom. Center dashed line is mean and error bands are standard deviation. Statistical significance was calculated by two-tailed Mann–Whitney U test at 15 min (ns P > 0.05, *P ≤ 0.05, ***P ≤ 0.0005). h–k 5HT2A overexpression in Cirl-/- showing that ectopic 5HT2A rescues MyoII levels in Cirl−/−. h MyoII images at different time points (left to right) in different genotypes (top to bottom): control (top panel), Cirl−/− (middle panel) and Cirl-/- 5HT2A + + (bottom panel). Quantification of time traces of MyoII intensities in DV and AP oriented junctions (i), amplitude of polarity (j), and medial MyoII intensities (k) in different genotypes. In the box plots in (c, d, f, i, j, k), the line in the middle is plotted at the median. The box extends from the 25th to 75th percentiles. The whiskers indicate minimum and maximum values. Statistics: ns P > 0.05, *P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.0005, ****P ≤ 0.00005 from two-tailed Mann–Whitney test. P values in (c): 1 (P = 0.635), 2 (P = 0.368), 3 (P = 0.031), 4 (P = 0.022), 5 (P = 0.011), 6 (P = 0.005), 7 (P = 0.0007), 8 (P = 0.0004), 9 (P = 0.005), 10 (P = 0.002), 11 (P = 0.93), 12 (P = 0.479), 13 (P = 0.669), 14 (P = 0.536), 15 (P = 0.475), 16 (P = 0.256), 17 (P = 0.126), 18 (P = 0.364), 19 (P = 0.417), 20 (P = 0.536). P values in (d): 1 (P = 0.098), 2 (P = 0.015), 3 (P = 0.007), 4 (P = 0.073), 5 (P = 0.031), 6 (P = 0.009), 7 (P = 0.004), 8 (P = 0.026), 9 (P = 0.026), 10 (P = 0.002), 11 (P = 0.015), 12 (P = 0.051), 13 (P = 0.035), 14 (P = 0.101), 15 (P = 0.073), 16 (P = 0.001), 17 (P = 0.004), 18 (P = 0.002), 19 (P = 0.002), 20 (P = 0.0002), 21 (P = 0.818), 22 (P = 0.628), 23 (P = 0.534), 24 (P = 0.445), 25 (P = 0.181). P values in (f): 1 (P = 0.012), 2 (P = 0.0004), 3 (P = 0.008), 4 (P = 0.088), 5 (P = 0.036), 6 (P = 0.088), 7 (P = 4.1 × 10−5), 8 (P = 1.65 × 10−4), 9 (P = 8.2 × 10−5), 10 (P = 2.88 × 10−4), 11 (P = 4.94 × 10−4), 12 (P = 4.94 × 10−4), 13 (P = 8.2 × 10−5), 14 (P = 8.2 × 10−5), 15 (P = 8.2 × 10−5), 16 (P = 1.65 × 10−4), 17 (P = 3.29 × 10−4), 18 (P = 8.2 × 10−5). P values in (i): 1 (P = 0.002), 2 (P = 0.01), 3 (P = 6.22 × 10−4), 4 (P = 0.005), 5 (P = 0.002), 6 (P = 0.003), 7 (P = 0.001), 8 (P = 0.005), 9 (P = 0.442), 10 (P = 0.505), 11 (P = 0.235), 12 (P = 0.2), 13 (P = 0.021), 14 (P = 0.021), 15 (P = 0.003), 16 (P = 0.05), 17 (P = 0.005), 18 (P = 0.002), 19 (P = 0.005), 20 (P = 0.002), 21 (P = 0.574), 22 (P = 0.442), 23 (P = 0.234), 24 (P = 0.195). P values in (j): 1 (P = 0.021), 2 (P = 0.021), 3 (P = 0.328), 4 (P = 0.13), 5 (P = 0.328), 6 (P = >0.999), 7 (P = 0.798), 8 (P = 0.878), 9 (P = 0.959), 10 (P = 0.005), 11 (P = 0.234), 12 (P = 0.105). P values in (k): 1 (P = 3.11 × 10−4), 2 (P = 1.55 × 10−4), 3 (P = 1.55 × 10−4), 4 (P = 1.55 × 10−4), 5 (P = 0.002), 6 (P = 0.001), 7 (P = 0.002), 8 (P = 0.001), 9 (P = 0.083), 10 (P = 0.028), 11 (P = 0.105), 12 (P = 0.234). n = number of embryos. Scale bars 5 µm. Source data are provided as a Source Data file.

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