Metabotropic glutamate receptors regulate differentiation of embryonic stem cells into GABAergic neurons

Metabotropic glutamate receptors regulate differentiation of embryonic stem cells into GABAergic neurons

Mouse embryonic stem (ES) cells were stimulated to differentiate either as adherent monolayer cultures in DMEM/F12 supplemented with N2/B27, or as floating embryoid bodies (EBs) exposed to 1  μ M retinoic acid (RA) for 4 days, starting from 4 DIV, and subsequently re-plated in DMEM/F12 medium. Adher...

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Bibliographic Details
Published in:Cell death and differentiation Vol. 15; no. 4; pp. 700 - 707
Main Authors: Sarichelou, I, Cappuccio, I, Ferranti, F, Mosillo, P, Ciceroni, C, Sale, P, Stocchi, F, Battaglia, G, Nicoletti, F, Melchiorri, D
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 01-04-2008
Nature Publishing Group
Subjects:
Acids
Aminobutyrates - pharmacology
Animals
Apoptosis
Benzopyrans - pharmacology
Biochemistry
Biomedical and Life Sciences
Cell Adhesion
Cell Biology
Cell Cycle Analysis
Cell death
Cell Differentiation - drug effects
Cell Line
Embryonic Stem Cells - drug effects
Embryonic Stem Cells - enzymology
Embryonic Stem Cells - metabolism
Excitatory Amino Acid Antagonists - pharmacology
gamma-Aminobutyric Acid - metabolism
Glutamate Decarboxylase - metabolism
Life Sciences
Membrane Potentials
Mice
Neurons - drug effects
Neurons - enzymology
Neurons - metabolism
original-paper
Phenotype
Physiology
Pyridines - pharmacology
Receptor, Metabotropic Glutamate 5
Receptors, Metabotropic Glutamate - drug effects
Receptors, Metabotropic Glutamate - metabolism
Stem Cells
Time Factors
Tretinoin - pharmacology
Tubulin - metabolism
Rome Italy
Italy
metabotropic glutamate receptors
neuronal differentiation
mouse embryonic stem cells
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Summary:Mouse embryonic stem (ES) cells were stimulated to differentiate either as adherent monolayer cultures in DMEM/F12 supplemented with N2/B27, or as floating embryoid bodies (EBs) exposed to 1  μ M retinoic acid (RA) for 4 days, starting from 4 DIV, and subsequently re-plated in DMEM/F12 medium. Adherent monolayer cultures of ES cells expressed mGlu5 receptors throughout the entire differentiation period. Selective pharmacological blockade of mGlu5 receptors with methyl-6-(phenylethynyl)-pyridine (MPEP) (1  μ M, added once a day) accelerated the appearance of the neuronal marker, β -tubulin. In addition, treatment with MPEP increased the number of cells expressing glutamate decarboxylase-65/67 (GAD 65/67 ), a marker of GABAergic neurons. In floating EBs, mGlu5 receptors are progressively replaced by mGlu4 receptors. The orthosteric mGlu4/6/7/8 receptor agonist, L -2-amino-4-phosphonobutanoate ( L -AP4), or the selective mGlu4 receptor enhancer, PHCCC, – both combined with RA at concentrations of 30  μ M – increased the expression of both β -tubulin and GAD 65/67 , inducing the appearance of fully differentiated neurons that released GABA in response to membrane depolarization. We conclude that mGlu receptor subtypes regulate neuronal differentiation of ES cells in a context-dependent manner, and that subtype-selective ligands of these receptors might be used for the optimization of in vitro protocols aimed at producing GABAergic neurons from ES cells.
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ISSN:1350-9047
1476-5403
DOI:10.1038/sj.cdd.4402298