ZFIN ID: ZDB-FIG-140311-16
McCarroll et al., 2013 - Fgf3 and Fgf10a work in concert to promote maturation of the epibranchial placodes in zebrafish. PLoS One   8(12):e85087 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 10-13 somites to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. 4

The anterior lateral line is the tissue source of Fgf10a responsible for facial placode development.

(A-D) confocal projections of Tg(pax2a:EGFP) zebrafish embryos (green) analyzed for Pax2a expression (magenta) at 14 (A), 18 (B), 21 (C), and 24 hpf (D). The presumptive facial placode is outlined in yellow as it condenses between 18 and 24 hpf. Insets show co-expression of Pax2a and Tg(pax2a:EGFP) at 14 hpf (A); by 18 hpf, however, Pax2a expression is absent in anterior lateral line precursors, while Tg(pax2a:EGFP) maintains expression in these cells (B). (E-E′′) Live confocal projection of a 12 hpf Tg(pax2a:Kaede) zebrafish embryo (E) with the anterior portion of the pax2a:Kaede+ domain photoconverted from green to red emission (E′) overlay (E′′). (F-F′′) Composite image of the same photoconverted embryo from (E) at 24 hpf analyzed for Pax2a expression (F) and cyan in (F′′) and photoconverted Kaede (F′) and red in (F′′). Note absence of Kaede positive cells in the facial placode. (G, H) In situ hybridization of eya1 in 52 hpf zebrafish embryos reveals proper neuromast deposition in control (G) and a failure of deposition and elongation of the anterior lateral line in fgf3-/-;fgf10-MO embryo (H; arrowheads). (I, J) Lateral views of the 24 hpf fgf3+10 morphant embryo showing the side that received wild-type donor cells (green) as well as the contralateral control side that did not receive donor cells. Pax2a expression is visualized by immunolabeling (magenta). Note partial rescue of the facial placode when wild-type donor cells were present in the presumptive anterior lateral line (J; arrowhead). (K, L) Quantification of Pax2a+ cells reveals a significant increase in the number of Pax2a+ cells in the facial (K) and glossopharyngeal and vagal placodes (L) of the transplanted sides versus contralateral sides (Wilcoxon matched-pairs signed rank test: **P<0.01; error bars: standard error of mean; n=8 embryos). Abbreviations: f, facial placode; g+v glossopharyngeal/vagal placode; ov, otic vesicle. Scale bars: 50 μm (A, I); 25 μm (G).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
eya1 AB control Long-pec anterior lateral line neuromast ISH
fgf3t26212/t26212 + MO3-fgf10a standard conditions Long-pec anterior lateral line ISH
GFP e1Tg standard conditions 10-13 somites anterior lateral line placode IHC
14-19 somites anterior lateral line placode IHC
14-19 somites Not Detected facial ganglion IHC
20-25 somites anterior lateral line placode IHC
20-25 somites Not Detected facial ganglion IHC
Prim-5 anterior lateral line placode IHC
Prim-5 Not Detected facial ganglion IHC
pax2a Ab1-pax2 e1Tg standard conditions 10-13 somites anterior lateral line placode IHC
Ab1-pax2 10-13 somites otic placode IHC
Ab1-pax2 14-19 somites Not Detected anterior lateral line placode IHC
Ab1-pax2 14-19 somites facial ganglion IHC
Ab1-pax2 20-25 somites Not Detected anterior lateral line placode IHC
Ab1-pax2 20-25 somites facial ganglion IHC
Ab1-pax2 Prim-5 Not Detected anterior lateral line placode IHC
Ab1-pax2 Prim-5 facial ganglion IHC
Ab1-pax2 nl12Tg standard conditions Prim-5 unspecified IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
fgf3t26212/t26212 + MO3-fgf10a standard conditions Long-pec anterior lateral line decreased object quality, abnormal
Long-pec anterior lateral line development decreased process quality, 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