title: Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers.
abstract: 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 modulations provide novel tools to understand the neural crest induction network.
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Plouhinec
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Roche
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Caterina
authors.initials: C
authors.lastname: Pegoraro
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Ana Leonor
authors.initials: AL
authors.lastname: Figueiredo
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Maczkowiak
authors.affiliation: University of California at Berkeley, Department of Molecular and Cell Biology and Center for Integrative Genomics, USA.
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Brunet
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Milet
authors.affiliation: Centre for Computational Biology, Mines ParisTech, 77300 Fontainebleau, France; Institut Curie, F-75248 Paris, France; INSERM U900, F-75248 Paris, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Vert
authors.affiliation: CNRS, iSSB, F-91000 Évry, France; Univ. Evry, iSSB, F-91000 Évry, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pollet
authors.affiliation: University of California at Berkeley, Department of Molecular and Cell Biology and Center for Integrative Genomics, USA.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Harland
authors.affiliation: Université Paris Sud, Centre Universitaire, F-91405 Orsay, France; Institut Curie, CNRS UMR3347, INSERM U1021, Centre Universitaire, F-91405 Orsay, France. Electronic address: anne-helene.monsoro-burq@curie.fr.
authors.forename: Anne H
authors.initials: AH
authors.lastname: Monsoro-Burq
mesh_terms: D000818:Animals; D024202:Electrophoretic Mobility Shift Assay; D018507:Gene Expression Regulation, Developmental / Q000235:genetics* / Q000502:physiology; D053263:Gene Regulatory Networks / Q000235:genetics* / Q000502:physiology; D017403:In Situ Hybridization; D046228:Microarray Analysis; D009432:Neural Crest / Q000196:embryology*; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism*; D029867:Xenopus Proteins / Q000378:metabolism*; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics
chemical_list: D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; C489411:Pax3 protein, Xenopus; D014157:Transcription Factors; D029867:Xenopus Proteins; C116546:Zic1 protein, Xenopus
keywords: Embryo; Gene regulatory network; Microarray; Neural crest; Pax3; Transcriptome; Xenopus laevis; Zic1