ZFIN ID: ZDB-FIG-190723-159
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:
Conditions:
Anatomical Terms:
Stage Range: 26+ somites to Long-pec
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: 26+ somites to Long-pec

Fig 4 Otic expression of <italic>fgf</italic> genes following mis-expression of <italic>fgf3</italic> or inhibition of Hh signalling.

(A–F’) In situ hybridisation for otic expression of fgf genes in Tg(hsp70:fgf3) embryos following a 30-minute heat shock (HS) at the 10-somite stage (14 hpf). Controls (left-hand panels of each pair of images) were sibling non-transgenic (Non-Tg) embryos subjected to the same heat shock. Numbers of embryos shown in the dorsal view panels indicate the number showing the phenotype from a mixed batch of transgenic and non-transgenic embryos in each pair of panels; 75% of the batch is expected to be transgenic. A–B’ show staining with a probe specific to the 3’ UTR of fgf3: note the ectopic patch of endogenous fgf3 expression at the posterior otic pole (B,B’; arrowheads) and disruption to fgf3 expression ventral to the otic vesicle (B’; asterisks) after heat shock in transgenic embryos. Expression of fgf10a is strengthened in the otic vesicle of transgenic embryos after heat shock (E–F’). (G–Q’) Expression of mRNA for fgf genes in embryos treated with 100 μM cyclopamine (cyc) from the 10-somite stage (14 hpf) until 22.5 hpf. Controls in the left-hand panels of each pair of images were treated with vehicle (ethanol) only. Numbers of embryos with the phenotype shown for individual treatments are indicated in the dorsal view panels. There was little change to the otic expression patterns of fgf3 or fgf8a at 22.5 hpf (8.5 hours post treatment) (G–J’), but note the loss of fgf3 expression ventral to the ear (H’; asterisk). Expression of fgf10a in the otic vesicle was strengthened after inhibition of Hh signalling in about 50% of treated embryos (L,L’). At 36 hpf (8.5 hpt + 13.5 h wash), ectopic expression of both fgf3 and fgf8a appeared in a new posteromedial domain in the ears of cyclopamine-treated embryos (M-P’; arrowheads). (Q,Q’) Expression of fgf8a in the posterior of the otic vesicle at 48 hpf (8.5 hpt + 25.5 h wash). Ectopic expression has strengthened (arrowhead) and medial epithelium is thinner than normal (brackets). Dorsal views of the left ear, with anterior to the top. Scale bar in A, 50 μm (applies to A–L); scale bar in A’, 50 μm (applies to A’–L’); scale bar in M, 50 μm (applies to M–P); scale bar in M’, 50 μm (applies to M’–P’); scale bar in Q, 20 μm (applies to Q,Q’).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
fgf3 AB control 26+ somites otic vesicle anterior region ISH
26+ somites pharyngeal endoderm ISH
Prim-25 otic vesicle anterior region ISH
AB chemical treatment by environment: Cyclopamine 26+ somites otic vesicle anterior region ISH
26+ somites Not Detected pharyngeal pouch ISH
Prim-25 otic vesicle anterior region ISH
Prim-25 otic vesicle posterior region ISH
AB heat shock 26+ somites otic vesicle anterior region ISH
26+ somites pharyngeal endoderm ISH
mitfaw2/w2 ; zf115Tg heat shock 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
26+ somites pharyngeal endoderm ISH
fgf8a AB control 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
Prim-25 otic vesicle anterior region ISH
Long-pec Not Detected otic vesicle posterior region ISH
AB chemical treatment by environment: Cyclopamine 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
Prim-25 otic vesicle anterior region ISH
Prim-25 otic vesicle posterior region ISH
Long-pec otic vesicle posterior region ISH
AB heat shock 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
mitfaw2/w2 ; zf115Tg heat shock 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
fgf10a AB control 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
AB chemical treatment by environment: Cyclopamine 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
AB heat shock 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
mitfaw2/w2 ; zf115Tg heat shock 26+ somites otic vesicle anterior region ISH
26+ somites otic vesicle posterior region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB chemical treatment by environment: Cyclopamine 26+ somites otic vesicle anterior region fgf10a expression increased amount, abnormal
26+ somites otic vesicle development decreased process quality, abnormal
26+ somites pharyngeal pouch fgf3 expression absent, abnormal
Prim-25 otic vesicle development decreased process quality, abnormal
Prim-25 otic vesicle posterior region fgf8a expression mislocalised, abnormal
Prim-25 otic vesicle posterior region fgf3 expression mislocalised, abnormal
Long-pec otic sensory epithelium postero-medial region decreased thickness, abnormal
Long-pec otic vesicle development decreased process quality, abnormal
Long-pec otic vesicle posterior region fgf8a expression increased amount, abnormal
Long-pec otic vesicle posterior region fgf8a expression mislocalised, abnormal
mitfaw2/w2 ; zf115Tg heat shock 26+ somites otic vesicle anterior region fgf10a expression increased amount, abnormal
26+ somites otic vesicle development decreased process quality, abnormal
26+ somites otic vesicle posterior region fgf10a expression increased amount, abnormal
26+ somites otic vesicle posterior region fgf3 expression mislocalised, abnormal
26+ somites pharyngeal endoderm fgf3 expression decreased amount, 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.