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. 2018 May 8;145(9):dev157487. doi: 10.1242/dev.157487

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© 2018. Published by The Company of Biologists Ltd

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Fig. 2. — Disrupted neural fold morphology and apical actin distribution in E9.5 Vangl2^+/Lp embryos, and in embryos treated with cytochalasin D and Y-27632. (A-L) Transverse sections through the PNP (see plane of the section in Fig. 1A) of Vangl2^+/+ (A-C), Vangl2^+/Lp (D-F), cytochalasin D-treated Vangl2^+/+ (G-I) and Y-27632-treated Vangl2^+/+ (J-L) embryos. (A,D,G,J) H&E staining; (B,C,E,F,H,I,K,L) phalloidin labelling. Arrowheads in A and B indicate the straight wild-type neural fold; note, by contrast, the outward flexion induced by mutation or treatment (D,E,G,H,J,K). Asterisk in B marks the curved DLHP of Vangl2^+/+ embryo; double asterisk in E marks the angle of the DLHP in the Vangl2^+/Lp embryos. White arrows in F,I,L point to disruption of the apical actin belt. (M,N) Dorsal views of phalloidin-stained flat-mount PNPs from Vangl2^+/+ and Vangl2^+/Lp embryos, respectively. (Oi-Oii) Magnification of the boxed areas in M and N, respectively. Yellow/red carets indicate cables of F-actin condensation on one side of aligned cells (mediolateral in Oi; anteroposterior in Oii); yellow arrowheads indicate cumuli of F-actin at one vertex of individual cells. (P) Orthogonal view of the cell marked in M by the yellow arrowhead, and outlined here, with a double asterisk marking the area of polarised actin cumuli. (Q) Diagram of cell alignment relative to the mediolateral axis of the embryo (0-45°, mediolateral orientation; 45-90°, anteroposterior orientation). (R) Proportion of cell alignments in anteroposterior (A-P; dark grey) or mediolateral (M-L; light grey) orientation (A-P orientation: 100% in Vangl2^+/+ vs 69.23% in Vangl2^+/Lp embryos: *P<0.05). (S) Number of cell alignments in Vangl2^+/+ (3.71 lines/embryo) and Vangl2^+/Lp (1.4/embryo) embryos. (Ti,Tii) Magnification of an area of phalloidin-stained PNT showing an actin cumulus in Vangl2^+/+ (Ti) and Vangl2^+/Lp (Tii) embryos. (U) Phalloidin intensity of the plot profile obtained from selected ROIs crossing the actin cumuli (solid bar) and basal actin at the linear boundary of two neighbouring cells (striped bar; yellow lines and purple lines, respectively, in Ti and Tii). (V) Graph of the number of actin cumuli in Vangl2^+/+ and Vangl2^+/Lp embryos. Error bars represent s.e.m. *P<0.05. Scale bars: 50 μm (A,B,D,E,G,H,J,K,M,N); 25 μm (C,F,I,L); 12.5 μm (Oi,Oii); 6.25 μm (Ti,Tii).