title: Gli1 is an inducing factor in generating floor plate progenitor cells from human embryonic stem cells.
abstract: Generation of mesencephalic dopamine (mesDA) neurons from human embryonic stem cells (hESCs) requires several stages of signaling from various extrinsic and intrinsic factors. To date, most methods incorporate exogenous treatment of Sonic hedgehog (SHH) to derive mesDA neurons. However, we and others have shown that this approach is inefficient for generating FOXA2+ cells, the precursors of mesDA neurons. As mesDA neurons are derived from the ventral floor plate (FP) regions of the embryonic neural tube, we sought to develop a system to derive FP cells from hESC. We show that forced expression of the transcription factor GLI1 in hESC at the earliest stage of neural induction, resulted in their commitment to FP lineage. The GLI1+ cells coexpressed FP markers, FOXA2 and Corin, and displayed exocrine SHH activity by ventrally patterning the surrounding neural progenitors. This system results in 63% FOXA2+ cells at the neural progenitor stage of hESC differentiation. The GLI1-transduced cells were also able to differentiate to neurons expressing tyrosine hydroxylase. This study demonstrates that GLI1 is a determinant of FP specification in hESC and describes a highly robust and efficient in vitro model system that mimics the ventral neural tube organizer.
authors.affiliation: Centre for Neuroscience, University of Melbourne, Parkville, Australia. mark.denham@unimelb.edu.au
authors.forename: Mark
authors.initials: M
authors.lastname: Denham
authors.forename: Lachlan H
authors.initials: LH
authors.lastname: Thompson
authors.forename: Jessie
authors.initials: J
authors.lastname: Leung
authors.forename: Alice
authors.initials: A
authors.lastname: Pébay
authors.forename: Anders
authors.initials: A
authors.lastname: Björklund
authors.forename: Mirella
authors.initials: M
authors.lastname: Dottori
mesh_terms: D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D004298:Dopamine / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D054259:Neural Tube / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051763:PAX7 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000071676:Zinc Finger Protein GLI1
chemical_list: D005136:Eye Proteins; C496503:Foxa2 protein, mouse; C400551:GLI1 protein, human; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051763:PAX7 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; C497322:Pax7 protein, mouse; D012097:Repressor Proteins; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1; D051542:Hepatocyte Nuclear Factor 3-beta; D004298:Dopamine
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