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ZFIN ID: ZDB-FIG-150820-28
Westcot et al., 2015 - Protein-Trap Insertional Mutagenesis Uncovers New Genes Involved in Zebrafish Skin Development, Including a Neuregulin 2a-Based ErbB Signaling Pathway Required during Median Fin Fold Morphogenesis. PLoS One   10:e0130688 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage: Long-pec

Fig. 5 nrg2a mutants display altered MFF morphology, consistent with the epidermal localization of the Nrg2-mRFP fusion protein and of endogenous nrg2a transcripts.

(A, B) By 48 hpf, nrg2amn0237Gt/mn0237Gt mutants (mn0237Gt/mn0237Gt) show altered MFF morphology. (A) Wild-type MFF edges are thin, flat, and continuously curved (arrowhead). (B) mn0237Gt/mn0237Gt mutant MFFs have thickened edges (arrowhead), and one or more pointed protrusions (open arrowhead). (C, D) Collagen II (Col II) immunostaining of actinotrichia support fibers within the MFF shows aberrant collagen accumulation and ectopic actinotrichia within mn0237Gt/mn0237Gt mutant apical MFFs (arrowheads) at 48 hpf. (E, G) At 24 hpf, Nrg2a-mRFP fusion protein is present in MFFs of heterozygous (+/mn0237Gt) embryos (E; view on tail of whole mount) and, at slightly lower levels, throughout the entire epidermis (G; section through tail region; immunostained for RFP and counterstained with DAPI). (F, H, I) Whole-mount in situ hybridization (WISH) demonstrates strong MFF expression of the GBT-generated fusion transcript (mRFP; F) in a representative +/mn0237Gt embryo at 24 hpf. When developed with Boehringer Blocking Reagent, WISH staining for endogenous nrg2a transcripts in 24 hpf wild-type embryos also revealed strong MFF expression of the endogenous gene (H, Boeringer). When developed without Boehringer Blocking Reagent, WISH staining further reveals uniform expression of the endogenous nrg2a gene throughout the entire epidermis (I). For cross-sections, see Honjo et al. (2008), Fig 6C [51]). (J-N) Co-labeling of a transverse section through the dorsal MFF of a +/mn0237Gt embryo at 24 hpf reveals restricted localization of the Nrg2a-mRFP fusion protein (J) in &916;Np63-positive basal keratinocytes (L), whereas the outer enveloping layer, labeled with EGFP (K), lacks the Nrg2a-mRFP protein; (M) DAPI counterstain; (N) merged image of different channels shown in (J-M).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
GFP nrg2amn0237Gt/+; gz7Tg standard conditions Prim-5 median fin fold periderm IHC
mRFP1 nrg2amn0237Gt/+ standard conditions Prim-5 epidermis IHC
Prim-5 epidermis ISH
Prim-5 median fin fold IHC
Prim-5 median fin fold ISH
nrg2amn0237Gt/+; gz7Tg standard conditions Prim-5 median fin fold keratinocyte IHC
Prim-5 Not Detected median fin fold periderm IHC
nrg2a WT standard conditions Prim-5 epidermis ISH
Prim-5 median fin fold ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-col2a IHC WT standard conditions Long-pec fin fold actinotrichium
IHC nrg2amn0237Gt/mn0237Gt standard conditions Long-pec fin fold actinotrichium
Ab1-tp63 IHC nrg2amn0237Gt/+; gz7Tg standard conditions Prim-5 median fin fold keratinocyte
Phenotype Details
Fish Conditions Stage Phenotype
nrg2amn0237Gt/mn0237Gt standard conditions Long-pec fin fold actinotrichium bent, abnormal
Long-pec fin fold actinotrichium increased thickness, abnormal
Long-pec fin fold actinotrichium mislocalised, abnormal
Long-pec median fin fold anatomical margin increased thickness, abnormal
Long-pec median fin fold anatomical margin protruding, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One