FIGURE

Fig. 5

ID
ZDB-FIG-050718-6
Publication
Wada et al., 2005 - Dual roles of zygotic and maternal Scribble1 in neural migration and convergent extension movements in zebrafish embryos
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Fig. 5

Identification of the llk gene. (A) Genetic map for the llk locus. The llk locus mapped to linkage group (LG) 7. The RDA marker, EcoRI-46 is located at 0 cM map-distance from the llk locus (0 per 2054 meioses). (B) Genomic structure of the zebrafish scrb1. The scrb1 gene is encoded by 45 exons spanning 120 kb in the genome. EcoRI-46 is located in the first intron of the gene. Each of the two mutant alleles, lklrw16 and llkrw468, carries a nucleotide substitution in exons 11 and 17, respectively. (C) Schematic drawing for the putative cDNA encoding 45 exons shown in B. Six alternatively used exons, 16, 28, 31, 34, 40 and 43 are shown as red boxes. Exon numbers and first ATG site are indicated above. Regions encoding LRR domain and four PDZ domains are indicated below. (D) RT-PCR analyses were performed to identify the predominant gene product. Primers were designed in the flanking exons encompassing each exon of interest. Total RNA was extracted from 1.5, 10, 18, 24, 36 and 48-hpf embryos. Arrowheads in each panel indicate the predominant RT-PCR product expressed during migration of the nVII motor neurons at 18-24 hpf. The predominant RT-PCR products contain exon 16, but no other exons (exons 28, 31, 34 and 43). The lesser RT-PCR products contain exons 28, 31, 34 and 43, but not exon 16 (indicated by arrows). 100 bp-interval molecular markers (bp) are shown in each panel. (E) Sequence diagrams of the mutation sites for the llkrw16 and llkrw468 alleles compared to the wild-type allele. (F) Schematic drawings of the wild-type (zScrb1) and mutant Scrb1 proteins (Scrbrw16 and Scrbrw468). Percentage identity of the amino acid sequences (%) to the mouse Scrb (mScrb1) is shown for each domain. The allele llkrw16 carries a mis-sense amino acid substitution in the first PDZ domain, while llkrw468 carries a stop codon in the LRR domain. (G-M) Lateral views of wild-type embryos stained with the scrb1 RNA probe in the 8-cell stage (G), dome-stage (H), 18 hpf (I), 22 hpf (J) and 48 hpf (K) embryos. (L,M) scrb1 mRNA expression in the brain at 20 hpf (L, dorsal view) and (M) cross section at r5 (indicated by the broken line in L). M′ shows the cross section at r5 of the 24 hpf-Isl1-GFP embryo stained with anti-acetylated α-tubulin antibody. Arrowheads indicate medial longitudinal fascicles (MLF); nc indicates notochord. Scale bars: 50 µm.

Expression Data
Gene:
Fish:
Anatomical Terms:
Stage Range: 16-cell to Long-pec

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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