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. 2023 Apr 8;26(5):106585. doi: 10.1016/j.isci.2023.106585

Figure 4.

Figure 4

Decrease in pushing force during fluid excretion

(A) Schematic of the experiment to release pushing force. The circumference (green line) and ventral side (magenta line) of the blastopore were excised, leaving the dorsal region.

(B and C) Control embryos with uncut ventral end (B), and embryos with pushing force released (C). Arrowhead, blastopore. Scale bars = 500 μm.

(D-I) Coronal section of the blastopore cleared with BABB. Wild-type embryos at stage 18 (D) and 20-21 (G). Control embryos with uncut ventral end at stage 18 (E) and 20-21 (H). Embryos with pushing force released at stage 18 (F) and 20-21 (I). Scale bars = 100 μm.

(J) Curvature of the CBC contact surface. Left and right columns show CBC curvature on the left and right sides, respectively. Gray lines correspond to the CBC of the same embryo. WT, wild-type embryo (stage 18, n = 12; stage 20-21, n = 12); Ctrl, control embryos (stage 18, n = 16; stage 20-21, n = 13); Rel., pushing force released embryo (stage 18, n = 13; stage 20-21, n = 14). ∗∗p = 0.0018, ∗∗∗p = 0.0007.

(K) Thickness of CBC at stage 18 (N = 24 CBCs from 12 embryos) and stage 20-21 (N = 24 CBCs from 12 embryos). Blue line; mean. ∗∗∗∗p < 0.0001.

(L) Contact length of the slit at stage 18 (n = 12) and stage 20-21 (n = 12). Blue line; mean. ∗∗∗∗p < 0.0001.

(M) Schematic of force measurement. MP, Measurement probe; PP, Pulling probe. Scale bars = 100 μm.

(N) Force required to open the blastopore at stage 18 (n = 35) and stage 20-21 (n = 49). Blue line; mean. ∗∗∗∗p < 0.0001. n = number of animals used. N = other data points. See also Figures S5 and S6 and Video S7.