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ZFIN ID: ZDB-FIG-171016-16
Ciarlo et al., 2017 - A chemical screen in zebrafish embryonic cells establishes that Akt activation is required for neural crest development. eLIFE   6 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-5
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: 14-19 somites to Prim-5

Fig. 2

CAPE decreases neural crest gene expression.

(A) CAPE dramatically reduces crestin:EGFP expression at 26 hpf. Figure 2—figure supplement 1 shows the response of a smaller (396 bp) crestin promoter fragment to CAPE and the timing of crestin response to CAPE. (B) CAPE decreases neural crest gene expression as determined by whole mount in situ hybridization (ISH) at 15–17 ss. Expression of some genes is decreased ubiquitously while others are only altered in specific anatomic locations (arrows). Images are representative of at least two independent experiments. (C) Scoring of ISH in (B). (D) CAPE reduces expression of pigment cell genes at 24 hpf. (E) FPKM values from RNA-sequencing of sox10:Kaede+ cells confirmed decreases in neural crest genes, though to a lesser extent than by ISH. Cells were sorted from control or CAPE-treated embryos at 17 ss. Figure 2—figure supplement 2 shows neural crest genes with no significant change by ISH. Figure 2—figure supplement 3 shows that a change in cell number does not account for neural crest gene expression changes. Figure 2—figure supplement 4 shows other gene expression changes in neural crest cells. *p<0.001, **p<0.0001, chi-square test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
crestin WT control 14-19 somites neural crest ISH
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest ISH
csf1ra WT control Prim-5 pigment cell ISH
WT chemical treatment by environment: phenethyl caffeate Prim-5 pigment cell ISH
dct WT control Prim-5 pigment cell ISH
WT chemical treatment by environment: phenethyl caffeate Prim-5 pigment cell ISH
dlx2a WT control 14-19 somites neural crest ISH
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest ISH
EGFP cz3334Tg control Prim-5 neural crest cell IFL
cz3334Tg chemical treatment by environment: phenethyl caffeate Prim-5 neural crest cell IFL
inka1a WT control 14-19 somites neural crest ISH
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest ISH
mitfa WT control Prim-5 pigment cell ISH
WT chemical treatment by environment: phenethyl caffeate Prim-5 pigment cell ISH
pax7a WT control 14-19 somites neural crest ISH
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest ISH
sox10 WT control 14-19 somites neural crest ISH
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: phenethyl caffeate 14-19 somites neural crest sox10 expression decreased amount, abnormal
14-19 somites neural crest dlx2a expression decreased amount, abnormal
14-19 somites neural crest crestin expression decreased amount, abnormal
14-19 somites neural crest inka1a expression decreased amount, abnormal
14-19 somites neural crest pax7a expression decreased amount, abnormal
Prim-5 pigment cell csf1ra expression decreased amount, abnormal
Prim-5 pigment cell mitfa expression decreased amount, abnormal
Prim-5 pigment cell dct expression decreased amount, abnormal
cz3334Tg chemical treatment by environment: phenethyl caffeate Prim-5 neural crest cell EGFP expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife