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ZFIN ID: ZDB-FIG-141209-7
Young et al., 2014 - The PDZ domain protein Mcc is a novel effector of non-canonical Wnt signaling during convergence and extension in zebrafish. Development (Cambridge, England)   141:3505-16 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 2-cell to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 1-4 somites to Prim-5

Fig. 3

ror2 is required for convergence and extension in zebrafish and acts upstream of mcc. (A) Schematic of the ror2 locus, with the position of splice junction MOs (MO1 and MO2) indicated. (B) Q-PCR analysis of ror2 expression in developing zebrafish. Maternal ror2 transcripts are detected from 2-256 cells, with dynamic expression over the next 3days. Data are normalized to β-actin. Error bars indicate s.e.m. epi, epiboly; s, somite. (C) ror2 WISH at the indicated stages. (D) Injection of ror2 MO1 results in CE defects at 10hpf, as assessed by ntl, dlx3 and hgg1 triple in situ hybridization; these defects are robustly rescued at 10hpf and 1dpf by mouse Ror2 mRNA. The solid black line with asterisk emphasizes the shortened yolk extension at 1dpf, and the curved dotted line highlights the smaller eyes and head in ror2 morphants compared with wild type. np, neural plate; p, polster; n, notochord. (E) Co-injection of ror2 MO1 and zebrafish mcc mRNA results in ostensibly normal embryos at 1dpf. (F,H) Membrane GFP-labeled notochord (F) and dorsal ectoderm (H) cells of wild type and mcc, ror2 or vangl2 morphants as indicated at 90-95% epiboly (9.5-10hpf). Dorsal view with anterior upwards. The notochord boundary in F is marked by vertical dashed yellow lines. Note the extra columns of cells in the morphant notochords. Dorsal ectoderm cells selected for morphometric analysis are outlined in H in white. The number of cells analyzed (n) and length-to-width (LWR) ratios (expressed as mean±s.e.m.) are indicated in the Rose diagrams, which depict cell orientation relative to the embryonic anterior-posterior axis (vertical dashed line). (G,I) The percentage of mediolaterally aligned cells with a longitudinal axis oriented ±20° with respect to the embryonic mediolateral axis (horizontal dashed line in the Rose diagrams). (D,E,F,H) MO and mRNA concentrations are provided in supplementary material Table S3, with scored embryo counts listed in supplementary material Table S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ctslb WT + MO3-ror2 standard conditions Bud polster ISH
dlx3b WT + MO3-ror2 standard conditions Bud preplacodal ectoderm ISH
mcc WT standard conditions 64-cell blastomere ISH
64-cell whole organism ISH
1k-cell blastomere ISH
1k-cell whole organism ISH
Shield whole organism ISH
Bud whole organism ISH
ror2 WT standard conditions 2-cell to 64-cell whole organism RTPCR
128-cell to 256-cell whole organism RTPCR
Oblong to Sphere whole organism RTPCR
50%-epiboly whole organism RTPCR
Shield whole organism RTPCR
1-4 somites to 5-9 somites whole organism RTPCR
5-9 somites whole organism RTPCR
10-13 somites whole organism RTPCR
14-19 somites whole organism RTPCR
20-25 somites whole organism RTPCR
Prim-5 whole organism RTPCR
Prim-15 whole organism RTPCR
Long-pec whole organism RTPCR
Protruding-mouth whole organism RTPCR
tbxta WT + MO3-ror2 standard conditions Bud notochord ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-mcc standard conditions 1-4 somites ectoderm cell orientation ectoderm, abnormal
1-4 somites ectoderm cell shape, abnormal
1-4 somites notochord cell orientation notochord, abnormal
1-4 somites notochord cell shape, abnormal
WT + MO1-vangl2 standard conditions 1-4 somites ectoderm cell orientation ectoderm, abnormal
1-4 somites ectoderm cell shape, abnormal
1-4 somites notochord cell orientation notochord, abnormal
1-4 somites notochord cell shape, abnormal
WT + MO3-ror2 standard conditions 1-4 somites ectoderm cell orientation ectoderm, abnormal
1-4 somites ectoderm cell shape, abnormal
1-4 somites notochord cell orientation notochord, abnormal
1-4 somites notochord cell shape, abnormal
Prim-5 extension decreased length, abnormal
Prim-5 eye decreased size, abnormal
Prim-5 head decreased size, abnormal
Prim-5 head malformed, abnormal
Prim-5 whole organism curved ventral, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development