title: Multiple roles of Activin/Nodal, bone morphogenetic protein, fibroblast growth factor and Wnt/β-catenin signalling in the anterior neural patterning of adherent human embryonic stem cell cultures.
abstract: Several studies have successfully produced a variety of neural cell types from human embryonic stem cells (hESCs), but there has been limited systematic analysis of how different regional identities are established using well-defined differentiation conditions. We have used adherent, chemically defined cultures to analyse the roles of Activin/Nodal, bone morphogenetic protein (BMP), fibroblast growth factor (FGF) and Wnt/β-catenin signalling in neural induction, anteroposterior patterning and eye field specification in hESCs. We show that either BMP inhibition or activation of FGF signalling is required for effective neural induction, but these two pathways have distinct outcomes on rostrocaudal patterning. While BMP inhibition leads to specification of forebrain/midbrain positional identities, FGF-dependent neural induction is associated with strong posteriorization towards hindbrain/spinal cord fates. We also demonstrate that Wnt/β-catenin signalling is activated during neural induction and promotes acquisition of neural fates posterior to forebrain. Therefore, inhibition of this pathway is needed for efficient forebrain specification. Finally, we provide evidence that the levels of Activin/Nodal and BMP signalling have a marked influence on further forebrain patterning and that constitutive inhibition of these pathways represses expression of eye field genes. These results show that the key mechanisms controlling neural patterning in model vertebrate species are preserved in adherent, chemically defined hESC cultures and reveal new insights into the signals regulating eye field specification.
authors.affiliation: Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Lupo
authors.forename: Claire
authors.initials: C
authors.lastname: Novorol
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Smith
authors.forename: Ludovic
authors.initials: L
authors.lastname: Vallier
authors.forename: Elena
authors.initials: E
authors.lastname: Miranda
authors.forename: Morgan
authors.initials: M
authors.lastname: Alexander
authors.forename: Stefano
authors.initials: S
authors.lastname: Biagioni
authors.forename: Roger A
authors.initials: RA
authors.lastname: Pedersen
authors.forename: William A
authors.initials: WA
authors.lastname: Harris
mesh_terms: D028341:Activins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002352:Carrier Proteins / Q000494:pharmacology; D002478:Cells, Cultured; D004149:Dioxoles / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D054258:Neural Plate / Q000378:metabolism; D055457:Nodal Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*; D000099176:Noggin Protein
chemical_list: D028341:Activins; D001549:Benzamides; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D004149:Dioxoles; D016222:Fibroblast Growth Factor 2; D005346:Fibroblast Growth Factors; D053823:Hedgehog Proteins; D055457:Nodal Protein; D051153:Wnt Proteins; D051176:beta Catenin; D000099176:Noggin Protein; C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide