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

I. Sarichelou, I. Cappuccio, F. Ferranti, P. Mosillo, C. Ciceroni, P. Sale, F. Stocchi, G. Battaglia, F. Nicoletti, D. Melchiorri

Research output: Contribution to journalArticlepeer-review


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 1μM 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 μM, 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 (GAD65/67), 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 μM - increased the expression of both β-tubulin and GAD65/67, 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.

Original languageEnglish
Pages (from-to)700-707
Number of pages8
JournalCell Death and Differentiation
Issue number4
Publication statusPublished - Apr 2008

ASJC Scopus subject areas

  • Cell Biology


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