NDLI: Directed Expression of Gata2, Mash1, and Foxa2 Synergize to Induce the Serotonergic Neuron Phenotype During In Vitro Differentiation of Embryonic Stem $Cells^{†‡§}$

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Concise review: Epidermal grafting: the case for pluripotent stem cells.

Conversion of Sox17 into a pluripotency reprogramming factor by reengineering its association with Oct4 on DNA.

Directed Expression of Gata2, Mash1, and Foxa2 Synergize to Induce the Serotonergic Neuron Phenotype During In Vitro Differentiation of Embryonic Stem $Cells^{†‡§}$

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Directed Expression of Gata2, Mash1, and Foxa2 Synergize to Induce the Serotonergic Neuron Phenotype During In Vitro Differentiation of Embryonic Stem $Cells^{†‡§}$

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Nefzger, Christian M. Haynes, John M. Pouton, Colin W.
Description	Country affiliation: Australia Author Affiliation: Nefzger CM ( Medicinal Chemistry and Drug Action, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Melbourne, Australia.)
Abstract	Investigation of serotonergic neuronal activity and its relationship to disease has been limited by a lack of physiologically relevant in vitro cell models. Serotonergic neurons derived from embryonic stem cells (ESCs) could provide a platform for such studies and provide models for use in drug discovery. Here, we report enhancement of serotonergic differentiation using a genetic approach. Expression of Gata2 increased the yield of serotonergic neurons. Enhancement was only achieved when Gata2 was expressed under the control of the tissue-specific promoter of the transcription factor Nkx6.1. High levels of Gata2 expression in ESCs compromised pluripotency and induced non-neuronal differentiation. Combined directed expression of Gata2, proneural gene Mash1, and forkhead transcription factor Foxa2 further enhanced serotonergic neural differentiation, resulting in a 10-fold increase in serotonin content. These neurons were also capable of depolarization (KCl, 30 mM)-induced elevations of intracellular $Ca^{2+}.$ The presence of sonic hedgehog during differentiation produced a further modest increase in numbers (1.5-fold). Transgene expression did not influence the number of tyrosine hydroxylase positive neurons in the cultures after 20 days, implying that Gata2, Mash1, and Foxa2 modulate in vitro differentiation at a time beyond the decision-point for dopaminergic or nondopaminergic commitment. This study demonstrates that the directed expression of specific transcription factors enhances serotonergic neuron differentiation in vitro and highlights the importance of transgene expression at the right stage of ESC differentiation to effect the generation of a desired neural subtype. STEM CELLS 2011;29:928–939
File Format	HTM / HTML
ISSN	10665099
Issue Number	6
Volume Number	29
e-ISSN	15494918
Journal	STEM CELLS
Language	English
Publisher	Wiley
Publisher Date	2011-06-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Cell Biology Discipline Embryology Basic Helix-loop-helix Transcription Factors Metabolism Cell Differentiation Embryonic Stem Cells Cytology Gata2 Transcription Factor Hepatocyte Nuclear Factor 3-beta Neurons Serotonin Biosynthesis Animals Genetics Cloning, Molecular Gata3 Transcription Factor Genes, Reporter Green Fluorescent Proteins Hedgehog Proteins Pharmacology Homeodomain Proteins Mice Peptide Elongation Factor 1 Phenotype Promoter Regions, Genetic Protein Biosynthesis Drug Effects Recombinant Fusion Proteins Beta-lactamases Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Medicine Molecular Medicine

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