Fig. 2
Non-canonical ligands Wnt5b and Wnt11 and PCP core components guide epithelial remodelling during chamber formation. a–c, e–k Top-down 0.5-μm confocal sections of hearts stained for Alcam (membrane marker) with the transition states in red and 3-point vertices in blue. aveTS, average number of transition states. a–c wt hearts at 26, 54 and 72 hpf. d Quantification of transition states per 100 cells in wt hearts. The LHT at 26 hpf exhibits 9.0 transitions/100 cells (n = 11). The two-chambered heart at 54 hpf shows 5.5 transitions/100 cells (n = 10), ***P = 0.0003, and at 72 hpf 4.5 transitions/100 cells (n = 6), 26 hpf vs. 72 hpf ****P < 0.0001, 54 hpf vs. 72 hpf P = 0.8157. Means ± s.d. Ordinary one-way ANOVA with Bonferroni’s multiple comparison test. Reported P values are multiplicity adjusted for each comparison. e–k Hearts at 54 hpf. Hearts of wnt5bta98 (e) and wnt11tx226 mutants (f) show a slight increase in transition states. In hearts of double wnt5bta98;wnt11tx226 mutants (g), the number of transition states increases significantly by 2-fold. The number of transition states is altered in hearts deficient in core PCP proteins (h–k). In hearts of fzd7ae3 mutants (h), and in hearts with reduced levels of dvl2 (i), the number of transition states significantly increases. In hearts of vangl2m209 mutants (j), and in pk1a-deficient hearts (k), the number of transition states mildly increases in comparison to wt hearts. l Quantification of average transition states/100 ventricular cardiomyocytes at 54 hpf. wt (5.5 transitions/100 cells, n = 10), wnt5bta98 (7 transitions/100 cells, n = 6), wnt11tx226 (6.8 transitions/100 cells, n = 5), wnt11ATG MO (6.4 transitions/100 cells, n = 5), wnt5bta98;wnt11tx226 (8.5 transitions/100 cells, n = 5, *P = 0.0489), wnt11ATG MO;wnt5bex6 MO (9.6 transitions/100 cells, n = 6, **P = 0.001), fzd7ae3 (8.8 transitions/100 cells, n = 7, **P = 0.0088), fzd7a5’UTR MO (10.63 transitions/100 cells, n = 9, ****P < 0.0001), dvl2ATG MO (8.5 transitions/100 cells, n = 7, *P = 0.0216), vangl2m209 (5.9 transitions/100 cells, n = 6), vangl2ATG MO (3.8 transitions/100 cells, n = 9), and pk1aATG MO (7.3 transitions/100 cells, n = 6). Means ± s.d. Ordinary one-way ANOVA with Bonferroni’s multiple comparison test. Reported P values are multiplicity adjusted for each comparison. Scale bars, 10 μm
