Fig. 1.
Mutations in the Wnt and PCP pathways affect the hair cell alignment in the lateral line. a Confocal image of a 5 days post fertilization (dpf) Tg(cldnb: lynGFP); Tg(sqet4); Tg(cldnb:H2A-mCherry) larva. b Schematic lateral view of a 5 dpf neuromast showing the different cell types. c Diagram of a 5 dpf larva showing the two different orientations of primI and primII-derived hair cells. d In situ hybridization of wnt11 (wnt11f1) in primI and primII-derived 5 dpf neuromasts. e–j Phalloidin stainings show hair cell orientations in primI-derived neuromasts of wild type (e), Wnt pathway mutants (f–h) and PCP mutants (i–j; Fisher’s Exact Test p-val for vangl2 primI = 7.33 × 10−28, MZscrib primI = 1.41 × 10−17). Individual hair cell orientation is depicted for each of the conditions tested. Black arrows denote disruption of the wild-type orientation. The Rose diagrams show the hair cell orientation distribution with respect to the longitudinal axis of the animal (horizontal) (WT n = 194 hair cells, MZwnt11 (wnt11f1) n = 353, gpc4 n = 222, MZfzd7a/7b n = 74, vangl2 n = 226, MZscrib n = 535). Bottom right depicts the Fisher’s exact test comparison with respect to the wild type for each condition. Top right shows the binomial test for each condition. k–p Phalloidin staining shows hair cell orientation in primII-derived neuromasts of wild type (k), Wnt pathway mutants (l–n Fisher’s Exact Test p-val for MZwnt11 (wnt11f1) primII = 1.69 × 10−23, gpc4 primII = 6.93 × 10−31, MZfzd7a/7b primII = 3.91 × 10−33) and PCP mutants (o–p, vangl2 primII p = 2.52 × 10−21, MZscrib primII = 9.22 × 10−12). Individual hair cell orientation is depicted for for each of the conditions tested and the color code is the same as in (e–j). The Rose diagrams reflect the same as in (e–j) (WT n = 209 hair cells, MZwnt11 (wnt11f1) n = 295, gpc4 n = 214, MZfzd7a/7b n = 182, vangl2 n = 141, MZscrib n = 392). For e–p, comparisons of angle distributions with respect to wild type, Fisher’s Exact Test ***p < 0.001, N.S not significant, binomial test A-P ***p < 0.001, A-P* = p < 0.05. e”–p”. A-P anteroposterior orientation, D-V dorsoventral orientation. Scale bar in a equals 500 μm, d equals 20 μm, e equals 5 μm