title: WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate.
abstract: Neural crest (NC) cells arise early in vertebrate development, migrate extensively and contribute to a diverse array of ectodermal and mesenchymal derivatives. Previous models of NC formation suggested derivation from neuralized ectoderm, via meso-ectodermal, or neural-non-neural ectoderm interactions. Recent studies using bird and amphibian embryos suggest an earlier origin of NC, independent of neural and mesodermal tissues. Here, we set out to generate a model in which to decipher signaling and tissue interactions involved in human NC induction. Our novel human embryonic stem cell (ESC)-based model yields high proportions of multipotent NC cells (expressing SOX10, PAX7 and TFAP2A) in 5 days. We demonstrate a crucial role for WNT/β-catenin signaling in launching NC development, while blocking placodal and surface ectoderm fates. We provide evidence of the delicate temporal effects of BMP and FGF signaling, and find that NC development is separable from neural and/or mesodermal contributions. We further substantiate the notion of a neural-independent origin of NC through PAX6 expression and knockdown studies. Finally, we identify a novel pre-neural border state characterized by early WNT/β-catenin signaling targets that displays distinct responses to BMP and FGF signaling from the traditional neural border genes. In summary, our work provides a fast and efficient protocol for human NC differentiation under signaling constraints similar to those identified in vivo in model organisms, and strengthens a framework for neural crest ontogeny that is separable from neural and mesodermal fates.
authors.affiliation: Kline Biology Tower, Department of Molecular, Cellular and Developmental Biology, Yale University, 266 Whitney Avenue, New Haven, CT 06511, USA Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06519, USA.
authors.forename: Alan W
authors.initials: AW
authors.lastname: Leung
authors.affiliation: Department of Biology, Eastern Connecticut State University, 83 Windham St., Willimantic, CT 06226, USA.
authors.forename: Barbara
authors.initials: B
authors.lastname: Murdoch
authors.affiliation: 203 School of Medicine Research Building, School of Medicine, University of California Riverside, Riverside, CA 92521, USA.
authors.forename: Ahmed F
authors.initials: AF
authors.lastname: Salem
authors.affiliation: 203 School of Medicine Research Building, School of Medicine, University of California Riverside, Riverside, CA 92521, USA.
authors.forename: Maneeshi S
authors.initials: MS
authors.lastname: Prasad
authors.affiliation: 203 School of Medicine Research Building, School of Medicine, University of California Riverside, Riverside, CA 92521, USA.
authors.forename: Gustavo A
authors.initials: GA
authors.lastname: Gomez
authors.affiliation: Kline Biology Tower, Department of Molecular, Cellular and Developmental Biology, Yale University, 266 Whitney Avenue, New Haven, CT 06511, USA 203 School of Medicine Research Building, School of Medicine, University of California Riverside, Riverside, CA 92521, USA martin.garcia-castro@ucr.edu.
authors.forename: Martín I
authors.initials: MI
authors.lastname: García-Castro
mesh_terms: D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D004475:Ectoderm / Q000166:cytology; D005136:Eye Proteins / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000166:cytology; D009432:Neural Crest / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D054258:Neural Plate / Q000166:cytology; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism
chemical_list: D019485:Bone Morphogenetic Proteins; D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D016212:Transforming Growth Factor beta; D051176:beta Catenin; D005346:Fibroblast Growth Factors
keywords: Embryonic stem cells; Human; Neural border; Neural crest; β-Catenin signaling
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