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Glutamine synthetase regulation by dexamethasone, RU486, and compound A in astrocytes derived from aged mouse cerebral hemispheres is mediated via glucocorticoid receptor

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Abstract

Glucocorticoids (GCs) regulate astrocyte function, while glutamine synthetase (GS), an enzyme highly expressed in astrocytes, is one of the most remarkable GCs-induced genes. GCs mediate their effects through their cognate glucocorticoid receptor (GRα and GRβ isoforms); however, the mechanism via which these isoforms regulate GS activity in astrocytes remains unknown. We used dexamethasone (DEX), a classical GRα/GRβ agonist, RU486, which is a specific GRβ ligand, and Compound A, a known “dissociated” ligand, to delineate the mechanism via which GR modulates GS activity. Aged Mouse Cerebral Hemisphere astrocytes were treated with DEX (1 μM), RU486 (1 nM–1 μM) or compound A (10 μM), alone or in combination with DEX. GS activity and expression, GR isoforms (mRNA and protein levels), and GRα subcellular trafficking were measured. DEX increased GS activity in parallel with GRα nuclear translocation. RU486 increased GS activity in absence of GRα nuclear translocation implicating thus a role of GRβ-mediated mechanism compound A had no effect on GS activity implicating a GRα–GRE-mediated mechanism. None of the compounds affected whole-cell GRα protein content. DEX reduced GRα and GRβ mRNA levels, while RU486 increased GRβ gene expression. We provide evidence that GS activity, in astrocytes, is regulated via GRα- and GRβ-mediated pathways with important implications in pathological conditions in which astrocytes are involved.

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Data availability

The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

U-2Os:

Human bone osteosarcoma epithelial cells

MEF cells:

Mouse embryonic fibroblast cells

IM-9:

Human B-lymphoblastoid cells

COS-1:

Fibroblast-like cell line from monkey kidney tissue

HCT116:

Human colon cancer cell line

FLS:

Fibroblast-like synoviocytes

CEM-C7:

Human T-lymphoblast cell line

BEAS-2B:

Human bronchial epithelial cell line

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Funding

This research was supported by the University Mental Health, Neurosciences and Precision Medicine Research Institute “Costas Stefanis” (UMHRI) and the Medical School, National and Kapodistrian University of Athens, University General Hospital “ATTIKON.”

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Authors and Affiliations

  1. 1st Department of Psychiatry, Eginition Hospital, Medical School, National and Kapodistrian University of Athens, Athens, Greece

    Theodosia Kazazoglou

  2. Cellular and Molecular Neurobiology Laboratory, University Mental Health, Neurosciences and Precision Medicine Research Institute “Costas Stefanis” (UMHRI), Athens, Greece

    Theodosia Kazazoglou

  3. Department of Clinical Biochemistry, Medical School, National and Kapodistrian University of Athens, University General Hospital “ATTIKON”, Rimini 1 str, Haidari, 12462, Athens, Greece

    Christina Panagiotou, Ioanna Kokkinopoulou, Anna Papadopoulou & Paraskevi Moutsatsou

  4. Department of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, Athens, Greece

    Chrysovalantou Mihailidou

Authors
  1. Theodosia Kazazoglou
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  2. Christina Panagiotou
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  3. Chrysovalantou Mihailidou
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  4. Ioanna Kokkinopoulou
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  5. Anna Papadopoulou
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  6. Paraskevi Moutsatsou
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Contributions

PM and TK contributed to the study concept and design. Material preparation, data collection, and analysis were performed by TK. Experiments were performed by TK, CP, CM, IK, and AP. Statistical analysis was carried out by CP. The first draft was written by TK. Writing and Review was performed by PM. All authors read and approved the final manuscript.

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Correspondence to Paraskevi Moutsatsou.

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Kazazoglou, T., Panagiotou, C., Mihailidou, C. et al. Glutamine synthetase regulation by dexamethasone, RU486, and compound A in astrocytes derived from aged mouse cerebral hemispheres is mediated via glucocorticoid receptor. Mol Cell Biochem 476, 4471–4485 (2021). https://doi.org/10.1007/s11010-021-04236-9

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  • DOI: https://doi.org/10.1007/s11010-021-04236-9

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