ZFIN Image: Kuan et al., 2015, Fig. 1

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Fig. 1

Isolation of mutant zebrafish wls alleles. (A–D) WT and (E–H) c186 homozygous mutants. (A, E) The c186 mutation was isolated through an RNA in situ hybridization screen using the asymmetrically expressed kctd12.1 gene. Dorsal views, 4 dpf. (B, F) Bright field images at 6 dpf. (C, G) Alcian blue staining of jaw cartilages and (D, H) pectoral fins at 6 dpf. (I) Genetic map of the chromosome 2 region where the two ENU induced lesions are located. The c186 allele is flanked by microsatellite markers z13620 and z60815 (recombination frequencies indicated by numbers in parentheses). Three genes were annotated in this interval: gadd45aa (i); gng12 (ii) and wls/gpr177 (iii). Nonsense mutations in the fh252 (C to T) and c186 (G to A) alleles reside in exons 3 and 7, respectively. (J–M) Rescue of mutant phenotypes by injection of wls mRNA. Lateral views of WT (J), homozygous c186 mutant (K) and rescued c186 mutant (L) TgBAC(gng8:Eco.NfsB-2A-CAAX-GFP) larvae at 7 dpf. The transgene labels habenular neurons and their axons with membrane-tagged GFP. (J′–L′) are dorsal views of the dorsal habenular nuclei. (M) Genotyping of larvae was used to confirm mutant rescue (refer to Section 4 for details).

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Acknowledgments

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Reprinted from Developmental Biology, 406(2), Kuan, Y.S., Roberson, S., Akitake, C.M., Fortuno, L., Gamse, J., Moens, C., Halpern, M.E., Distinct requirements for Wntless in habenular development, 117-28, Copyright (2015) with permission from Elsevier. Full text @ Dev. Biol.