ZFIN The Zebrafish Information Network (original) (raw)

Figure 6 of Derrick et al., 2023

Asymmetric spaw expression in vangl2m209 mutants cannot be rescued by VANGL2 variants. (A–A″) Representative images of mRNA in situ hybridization for southpaw (spaw) the zebrafish homologue of Nodal at 20 hpf. In wild-type siblings (A), spaw is restricted to the left lateral plate mesoderm (LPM, arrowhead), alternatively, spaw may be abnormally expressed in the right LPM (A′, arrowhead) or bilaterally (A″, arrowheads). (B) Quantification of abnormal spaw expression at 20 hpf to examine efficacy of WT VANGL2 to rescue homozygous vangl2m209 mutant phenotype. WT VANGL2 mRNA injection completely rescues spaw expression. Colour-coded regions defined by vangl2m209 mutant data range: within/no rescue is grey, below/rescue is green and above/worsens is magenta. Each dot denotes a clutch, consisting of 17–30 embryos. (C) Quantification of abnormal spaw expression at 20 hpf in homozygous vangl2m209 mutants injected with mRNA encoding different VANGL2 VUS. Variants are ranked and grouped based on result of statistical test when compared to uninjected homozygous vangl2m209 mutants and homozygous vangl2m209 mutants injected with WT VANGL2 mRNA (see Supplementary Table 5 for statistical tests). Each dot denotes a clutch, consisting of 18–43 embryos, colour coded regions and dots denote comparison against the uninjected homozygous vangl2m209 mutant data from panel B. A–A″: Dorsal views, anterior up. B-C: One-way ANOVA with multiple comparisons, Mean ± SEM, ****: p < 0.0001.

Fig. 3 of Banerjee et al., 2024

vps16(-/-) mutants show significant cell death and hypomyelination by 7 dpf. (A–I) TUNEL assay performed on brain sections of 7 dpf sibling and vps16(-/-) mutant larvae. Brain sections identified and sub-categorized in four regions: (A,B) forebrain, (C,D) fore-midbrain, (E,F) mid-hindbrain, and (G,H) hindbrain. (A,C,E,G) Minimal to no TUNEL-positive apoptotic cells (green) seen in sibling brain regions (n: 10 forebrain, 7 fore-midbrain, 9 mid-hindbrain, 4 hindbrain). (B,D,F,H) Numerous TUNEL-positive apoptotic cells (green) seen in vps16(-/-) mutant brain regions (n = 10 forebrain, p = 0.0022; n = 7 fore-midbrain, p = 0.0005; n = 5 mid-hindbrain, p = 0.0146; n = 7 hindbrain, p = 0.0027). (I) Graph representing quantification of average number of TUNEL-positive apoptotic cells in specific brain regions of 7 dpf siblings and vps16(-/-) mutants. (J–N) Immunolocalization of MBP (myelin, green) and HuCD (neurons, red) in brain section from two regions of sibling and vps16(-/-) mutant larvae: (J,K) mid-hindbrain and (L,M) hindbrain. (J–L) MBP immunolocalizes to Mauthner axons in mid-hindbrain (n = 6) and hindbrain (n = 6) regions of sibling brain. (K,M) MBP immunolocalization in mid-hindbrain (n = 6, p = 0.0048) and hindbrain (n = 6, p = 0.0061) regions of vps16(-/-) mutant brain showing decrease in myelin content. (N) Graph representing quantification of MBP immunolocalization in specific brain regions of 7 dpf siblings and vps16(-/-) mutants. Scale bar in panel A = 50 microns for all images. Blue = TO-PRO-3, nuclear stain. Error bars indicate SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.