Endogenous activation of mGlu5 metabotropic glutamate receptors supports self-renewal of cultured mouse embryonic stem cells

Cappuccio, I.; Spinsanti, Paola; Porcellini, Antonio; Desiderati, Francesca; Vita, Teresa De; Storto, Marianna; Capobianco, Loredana; Battaglia, Giuseppe; Nicoletti, Ferdinando; Melchiorri, Daniela

doi:10.1016/j.neuropharm.2005.05.014

Cited by 49 publications

(58 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We moved from the findings that (i) undifferentiated ES cells cultured in the presence of LIF express mGlu5 receptors, but not other mGlu receptor subtypes; 16 (ii) pharmacological blockade of mGlu5 receptors induces differentiation towards the neuroectoderm lineage; 16 (iii) in ES cells differentiating into EBs, mGlu5 receptors are progressively replaced by mGlu4 receptors 18 and (iv) activation of mGlu4 receptors enhances the expression of early neural markers in EBs treated with RA. 18 To further investigate the role of mGlu5 and mGlu4 receptors in ES cell differentiation, we used two different differentiation protocols: (i) cultures of adherent ES cells exposed to serum-free N2B27 medium, which favours neuronal differentiation 19 and (ii) floating cultures of EBs exposed to all-trans-RA (1 mM) according to the 4À/4 þ protocol (i.e.…”

Section: Resultsmentioning

confidence: 99%

“…in the presence of LIF). 16 Here, we have shown that ES cells maintain the expression of mGlu5 receptors when they are forced to differentiate into neurons by switching the growing medium into a specific chemically defined medium (the N2/B27 medium) lacking both serum and LIF. Under these conditions, mGlu5 receptor blockade with MPEP increased the rate of appearance of the neuronal marker, b-tubulin, and increased the number of putative GABAergic neurons immunopositive for the GAD.…”

Section: Discussionmentioning

confidence: 96%

“…Previous studies have shown that the mGlu5 receptor is the only mGlu receptor subtype expressed by undifferentiated ES cells, and that endogenous activation of mGlu5 receptors sustains self-renewal of these cells. 16,17 Pharmacological blockade of mGlu5 receptors promotes differentiation of ES cells towards the neuroectoderm lineage even under proliferating conditions (i.e. in the presence of LIF).…”

Section: Discussionmentioning

confidence: 99%

“…20,21 This is particularly relevant to this model because ES cells release micromolar amounts of glutamate in the medium. 16 As an alternative differentiation protocol, ES cells were plated onto a non-adhesive substrate in a serum-containing medium deprived of LIF. In this particular case, ES cells give rise to cells of EBs, which can be further differentiated into neurons by the addition of RA followed by re-plating in a DMEM/F12/ N2 medium.…”

Section: Discussionmentioning

confidence: 99%

“…We have found that cultured mouse embryonic stem (ES) cells express mGlu5 receptors, the activation of which supports the selfrenewal of ES cells in cooperation with leukemia inhibitory factor (LIF). 16,17 Pharmacological blockade or knockdown of mGlu5 receptors in ES cells cultured in the presence of LIF drives cell differentiation towards the neuroectoderm lineage. When ES cells are induced to differentiate into cells of the three germ layers by the formation of embryoid bodies (EBs), mGlu5 receptors are replaced by mGlu4 receptors.…”

mentioning

confidence: 99%

See 4 more Smart Citations

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

et al. 2008

View full text Add to dashboard Cite

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 lM 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 lM, added once a day) accelerated the appearance of the neuronal marker, b-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 lM -increased the expression of both b-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.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

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

et al. 2008

View full text Add to dashboard Cite

show abstract

Amino Acid Transporters: Roles for Nutrition and Signalling in Embryonic and Induced Pluripotent Stem Cells

Winkle

2013

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Amino acids serve both to nourish and signal in cells and, consequently, so do their biomembrane transporters. In fact, some of these transporters may initiate signalling while transporting an amino acid substrate rather than serving simply to transport a signalling molecule. Most amino acid transporters now appear to have been cloned, and virtually all of the cloned transporters are listed in solute carrier tables for easy access online ( http://slc.bioparadigms.org/ ). It is more difficult to decide as to which transporters are expressed in a given tissue, and their transport and signalling functions will likely continue to emerge long after all of the transporters have been identified. These ongoing investigations of transporter identification and function for embryonic and induced pluripotent stem cells are illustrated. Surprisingly, characterisation of amino acid transport in these cells is in its infancy. Thus, they serve as an experimental model for investigating the expression of known and novel transporters. Key Concepts: At least 55 genes encode amino acid transporters in a minimum of 10 gene families (see tables at http://slc.bioparadigms.org/ ). An amino acid may be listed as a substrate for a cloned transporter in these tables even though its transport has not been studied in detail. The characteristics of cloned amino acid transporters may not correspond to the ways they function in cellular physiology. Stem cells serve as an experimental model for investigating the expression of known and novel transporters. Threonine uptake and glutamate exodus from stem cells via transporters each fosters cellular proliferation. Glutamate exodus from stem cells is needed to stimulate glutamate receptors. Proline uptake by stem cells promotes their differentiation. mTOR signalling in stem cells is necessary for both proline‐induced differentiation and threonine‐induced proliferation. Both threonine and glutamate promote stem cell proliferation by enhancing the expression of cMyc. Threonine and glutamate likely foster proliferation in human as well as mouse stem cells.

show abstract

Ca²⁺‐depended signaling pathways regulate self‐renewal and pluripotency of stem cells

Ермаков

Daks

Fedorova

et al. 2018

Cell Biology International

View full text Add to dashboard Cite

Ca -mediated signaling is widely spread in nature and plays critical role in the individual development of various organisms ranging from microorganisms to mammals. In vertebrates, Ca is involved in important developmental events: fertilization, body plan establishment, and organogenesis. The two later events are defined by embryonic stem cells (ESCs). ESCs are capable of self-renewal and are pluripotent by nature, that is, can give rise to all types of cells that make up the body. Given the paramount importance of Ca signalization in the development, it is therefore not surprising this process also plays role in the biology of stem cells. In this review, we scrutinize the published experimental data on the role of Ca ions in embryonic stem cells self-renewal and pluripotency. In line with this, we also discuss possible mechanisms of p53 inhibition as a major hindrance to self-renewal of ESCs. Finally, we argue about the role of G-protein-coupled receptors (GPCRs), the largest family of heteromeric transmembrane receptors, and GPCR-mediated signalization in stem cells, and propose the role for the GPCR-G-protein-PLC-Ca -downstream signaling pathway in the regulation of pluripotency of both mouse and human ESCs.

show abstract

Endogenous activation of mGlu5 metabotropic glutamate receptors supports self-renewal of cultured mouse embryonic stem cells

Cited by 49 publications

References 38 publications

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

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

Amino Acid Transporters: Roles for Nutrition and Signalling in Embryonic and Induced Pluripotent Stem Cells

Ca²⁺‐depended signaling pathways regulate self‐renewal and pluripotency of stem cells

Contact Info

Product

Resources

About

Endogenous activation of mGlu5 metabotropic glutamate receptors supports self-renewal of cultured mouse embryonic stem cells

Cited by 49 publications

References 38 publications

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

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

Amino Acid Transporters: Roles for Nutrition and Signalling in Embryonic and Induced Pluripotent Stem Cells

Ca2+‐depended signaling pathways regulate self‐renewal and pluripotency of stem cells

Contact Info

Product

Resources

About

Ca²⁺‐depended signaling pathways regulate self‐renewal and pluripotency of stem cells