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| Status |
Public on Dec 28, 2013 |
| Title |
Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers [X_laevis_2] |
| Organism |
Xenopus laevis |
| Experiment type |
Expression profiling by array
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| Summary |
Neural crest development is orchestrated by a complex and still poorly understood gene regulatory network. Premigratory neural crest is induced at the lateral border of the neural plate by the combined action of signaling molecules and transcription factors such as AP2, Gbx2, Pax3 and Zic1. Among them, Pax3 and Zic1 are both necessary and sufficient to trigger a complete neural crest developmental program. However, their gene targets in the neural crest regulatory network remain unknown. Here, through a transcriptome analysis of frog microdissected neural border, we identified an extended gene signature for the premigratory neural crest, and we defined novel potential members of the regulatory network. This signature includes 34 novel genes, as well as 44 known genes expressed at the neural border. Using another microarray analysis which combined Pax3 and Zic1 gain-of-function and protein translation blockade, we uncovered 25 Pax3 and Zic1 direct targets within this signature. We demonstrated that the neural border specifiers Pax3 and Zic1 are direct upstream regulators of neural crest specifiers Snail1/2, Foxd3, Twist1, and Tfap2b. In addition, they may modulate the transcriptional output of multiple signaling pathways involved in neural crest development (Wnt, Retinoic Acid) through the induction of key pathway regulators (Axin2 and Cyp26c1). We also found that Pax3 could maintain its own expression through a positive autoregulatory feedback loop. These hierarchical inductions, feedback loops, and pathway modulation provide novel tools to understand the neural crest induction network.
Transcriptomes of animal caps overexpressing Pax3+/-Zic1 in the presence/absence of cycloheximide, a translation inhibitor, were compared to control animal caps to identify direct Pax3 and Zic1 targets
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| Overall design |
2-4 cells stage embryos were injected with inducible Pax3-GR+/-Zic1-GR constructs. Animal caps were cut at stage 9. Cycloheximide (Chx, 0.1mg/ml) was then applied to the healed animal caps, from stage 10 to 10.5 (i.e. for 30 min at 23*C), then dexamethasone (Dex) was added at stage 10.5 to the cycloheximide-containing medium. Explants were rinced and lysed after two additional hours at 23*C, i.e. when sibling embryos reached stage 11.5-12. Total RNA was then extracted and hybridized on Affymetrix microarrays. Transcriptomes were compared to determine Pax3 and Zic1 targets.
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| Contributor(s) |
Plouhinec J, Roche DD, Pegoraro C, Figueiredo A, Maczkowiak F, Brunet LJ, Vert J, Pollet N, Harland RM, Monsoro-Burq AH |
| Citation(s) |
24360906 |
| Submission date |
Dec 27, 2013 |
| Last update date |
Oct 02, 2015 |
| Contact name |
Anne-Helene Monsoro-Burq |
| E-mail(s) |
Anne-Helene.Monsoro-Burq@curie.fr
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| Organization name |
Institut Curie
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| Department |
Université Paris Sud
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| Lab |
U1021 - UMR3347
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| Street address |
Bat 110
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| City |
Orsay |
| ZIP/Postal code |
91405 |
| Country |
France |
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| Platforms (1) |
| GPL10756 |
[X_laevis_2] Affymetrix Xenopus laevis Genome 2.0 Array |
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| Samples (16)
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| GSM1298762 |
Control animal cap, stage 12, replicate 1 |
| GSM1298763 |
Pax3-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 1 |
| GSM1298764 |
Pax3-injected animal cap, stage 12, induced with Dex, replicate 1 |
| GSM1298765 |
Pax3Zic1-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 1 |
| GSM1298766 |
Pax3Zic1-injected animal cap, stage 12, induced with Dex, replicate 1 |
| GSM1298767 |
Control animal cap, stage 12, replicate 2 |
| GSM1298768 |
Pax3-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 2 |
| GSM1298769 |
Pax3-injected animal cap, stage 12, induced with Dex, replicate 2 |
| GSM1298770 |
Pax3Zic1-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 2 |
| GSM1298771 |
Pax3Zic1-injected animal cap, stage 12, induced with Dex, replicate 2.1 |
| GSM1298772 |
Pax3Zic1-injected animal cap, stage 12, induced with Dex, replicate 2.2 |
| GSM1298773 |
Control animal cap, stage 12, replicate 3 |
| GSM1298774 |
Pax3-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 3 |
| GSM1298775 |
Pax3-injected animal cap, stage 12, induced with Dex, replicate 3 |
| GSM1298776 |
Pax3Zic1-injected animal cap, stage 12, treated with Chx, induced with Dex, replicate 3 |
| GSM1298777 |
Pax3Zic1-injected animal cap, stage 12, induced with Dex, replicate 3 |
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| This SubSeries is part of SuperSeries: |
| GSE53679 |
Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers |
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| Relations |
| BioProject |
PRJNA232629 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE53677_RAW.tar |
46.5 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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