ZFIN ID: ZDB-FIG-190723-160
Hartwell et al., 2019 - Anteroposterior patterning of the zebrafish ear through Fgf- and Hh-dependent regulation of hmx3a expression. PLoS Genetics   15:e1008051 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Protruding-mouth

Fig 5 Fused otoliths and sensory maculae, and reduction of anterior otic character, in <italic>hmx3a</italic><sup><italic>SU3/SU3</italic></sup> mutants.

(A) Schematic diagram showing the predicted truncated product for the hmx3aSU3 allele. The mutation was generated using a CRISPR sgRNA targeting sequence in exon 2 upstream of the DNA-binding homeodomain (green). The predicted truncated protein produced by the hmx3aSU3/SU3 allele contains a Thr to Gly substitution at amino acid 107, followed by a stretch of 10 further incorrect amino acids (magenta). The truncated protein lacks the homeodomain. (B,C) Differential interference contrast (DIC) images of ears in live embryos at 3 dpf (72 hpf). Numbers of embryos in a batch from a mating between heterozygous parents are given. Note the fused otolith in the hmx3aSU3/SU3 mutant ear (C). (D–E’) FITC-phalloidin stains of the sensory maculae (D,E) and cristae (D’,E’) in the ear at 3 dpf (72 hpf). Numbers of embryos showing the phenotype from a cross between heterozygous parents are shown. White arrowhead: anterior macula; white arrow: posterior macula; asterisks indicate cristae. Additional examples are shown in S9 Fig. (F–M’) In situ hybridisation for otic anterior markers at 24 hpf in genotyped wild-type and hmx3aSU3/SU3 mutant embryos. The dotted outline marks the outer edge of the otic epithelium. Black arrowheads in F–I indicate the extent of hmx expression in medial epithelium; white arrowheads indicate areas of reduced expression levels; blue arrowhead in F’ marks presumed otic or anterior lateral line neuroblasts; light blue arrowhead in G’ indicates loss of expression in this area; red arrowheads in G’,I’ mark expansion of expression in ventral otic epithelium. Black arrowhead in L’ indicates anterior otic expression domain of fgf3, lost in M,M’ (white arrowheads); white double-headed arrows mark expression of fgf3 in pharyngeal pouch endoderm. Numbers in panels F–M indicate numbers of embryos genotyped as either wild type or homozygous mutant that showed the representative expression patterns illustrated. Scale bars, 50 μm (scale bar in B applies to B,C; scale bar in D applies to D–E’; scale bar in F applies to F–I, J–M; scale bar in F’ applies to F’–I’, J’–M’).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
fgf3 AB standard conditions Prim-5 otic vesicle anterior region ISH
Prim-5 pharyngeal pouch ISH
hmx3asu3/su3(AB) standard conditions Prim-5 otic vesicle anterior region ISH
Prim-5 pharyngeal pouch ISH
hmx2 AB standard conditions Prim-5 otic vesicle anterior region ISH
hmx3asu3/su3(AB) standard conditions Prim-5 otic vesicle anterior region ISH
hmx3a AB standard conditions Prim-5 otic vesicle anterior region ISH
Prim-5 statoacoustic (VIII) ganglion neuroblast ISH
hmx3asu3/su3(AB) standard conditions Prim-5 otic vesicle anterior region ISH
Prim-5 otic vesicle postero-medial region ISH
Prim-5 statoacoustic (VIII) ganglion neuroblast ISH
pax5 AB standard conditions Prim-5 otic vesicle anterior region ISH
hmx3asu3/su3(AB) standard conditions Prim-5 otic vesicle anterior region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
hmx3asu3/su3(AB) standard conditions Prim-5 otic vesicle anterior region fgf3 expression decreased amount, abnormal
Prim-5 otic vesicle anterior region hmx2 expression decreased amount, abnormal
Prim-5 otic vesicle anterior region hmx3a expression decreased amount, abnormal
Prim-5 otic vesicle anterior region pax5 expression decreased amount, abnormal
Prim-5 otic vesicle postero-medial region hmx3a expression mislocalised, abnormal
Prim-5 statoacoustic (VIII) ganglion neuroblast hmx3a expression decreased amount, abnormal
Protruding-mouth inner ear has normal numbers of parts of type anterior crista, normal
Protruding-mouth inner ear has normal numbers of parts of type lateral crista, normal
Protruding-mouth inner ear has normal numbers of parts of type posterior crista, normal
Protruding-mouth inner ear morphogenesis decreased process quality, abnormal
Protruding-mouth lapillus fused with sagitta, abnormal
Protruding-mouth posterior macula fused with anterior macula, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.