ZFIN Figure: Navajas Acedo et al., 2019, Fig. 5

Fig. 5

Wnt pathway genes are required for proper coordinated support cell organization. a, b Phalloidin staining of wild-type primI and primII-derived neuromasts. The dotted square outlines the area magnified in (a) and (b). Two support cells each were false colored to depict their orientation. b–j Phalloidin staining of neuromasts 3 h after hair cell ablation. c, d Wild-type primI (c) and primII-derived (d) neuromasts. e, f vangl2 mutant primI (e) and primII-derived (f) neuromasts. g, h MZscrib mutant primI (g) and primII-derived (h) neuromasts. i, j MZwnt11 (wnt11f1) mutant primI (i) and primII-derived (j) neuromasts. k, l vangl2;MZwnt11 (wnt11f1) double mutant primI (k) and primII-derived (l) neuromasts. The histograms show the distribution of binned angles of cell orientation with respect to the horizontal for each of the conditions tested. c, d Angle distribution in wild-type primI (c one-way chi square test p = 1.78 × 10⁻⁸⁷) and primII-derived (d p = 4.54 × 10⁻¹⁹) neuromasts. Note that primI distribution is skewed toward being horizontally aligned, while the distribution in primII-derived neuromasts is toward vertical alignment. e, f Angle distribution in vangl2 mutant primI (e p = 4.91 × 10⁻¹³) and primII-derived (f p = 2.14 × 10⁻¹⁶) neuromasts. g, h Angle distribution in MZscrib mutant primI (g p = 1.40 × 10⁻¹⁴) and primII-derived (h p = 9.60 × 10⁻¹³) neuromasts. i, j Support cell orientation in MZwnt11 (wnt11f1) mutant primI (i p = 5.32 × 10⁻⁵²) and primII-derived (j p = 0.006) neuromasts. k, l Support cell orientation in vangl2;MZwnt11 (wnt11f1) double mutant primI- (k p = 6.69 x 10^-26) and primII-derived (l p = 0.22) neuromasts. m, n Double β-II-Spectrin (labeling the cuticular plates) and ZO-1 staining of a 5 dpf wild-type neuromasts. o, p Wild-type atoh1a CRISPants do not possess hair cells (β-II-spectrin is absent; see also Supplementary Fig. 5) and the support cells in all neuromasts show coordinated orientation (one-way chi-square test p-value in o = 4.98 × 10⁻²⁰, p = 2.63 × 10⁻¹⁹). q, r MZwnt11 (wnt11f1) injected with atoh1a CRISPR show coordinated support cell orientation in primI-derived neuromasts (q p = 2.88 × 10⁻⁶⁰) and do not show coordinated support cell orientation in primII neuromasts (r p = 0.24). Scale bar in a, c, and o equal 5 μm

Expression Data

Antibodies:	Ab1-sptbn1 Ab1-tjp1
Fish:	WT + CRISPR2-atoh1a + CRISPR3-atoh1a wnt11^fh224/fh224 + CRISPR2-atoh1a + CRISPR3-atoh1a
Knockdown Reagents:	CRISPR2-atoh1a CRISPR3-atoh1a
Anatomical Term:	posterior lateral line neuromast cell-cell junction
Stage:	Day 5

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	scrib^rw468/rw468 vangl2^m209/m209 wnt11^fh224/fh224 WT + CRISPR2-atoh1a + CRISPR3-atoh1a wnt11^fh224/fh224 + CRISPR2-atoh1a + CRISPR3-atoh1a wnt11^fh224/fh224; vangl2^m209/m209
Condition:	chemical treatment by environment: neomycin
Knockdown Reagents:	CRISPR2-atoh1a CRISPR3-atoh1a
Observed In:	anterior-posterior polarized posterior lateral line neuromast cuticular plate anterior-posterior polarized posterior lateral line neuromast neuromast hair cell anterior-posterior polarized posterior lateral line neuromast neuromast support cell dorsal-ventral polarized posterior lateral line neuromast cuticular plate dorsal-ventral polarized posterior lateral line neuromast neuromast hair cell dorsal-ventral polarized posterior lateral line neuromast neuromast support cell
Stage:	Day 5

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.