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Cell type-specific in vivo expression of genes encoding signalling molecules in the brain in response to chronic mild stress and chronic treatment with fluoxetine

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Abstract

Rationale

Previously, we reported that chronic treatment with fluoxetine increased gene expression of 5-hydroxytryptamine receptor 2B (5-HT2BR), cytosolic phospholipase 2α (cPLA2α), glutamate receptor, ionotropic kainate 2 (GluK2) and adenosine deaminase acting on RNA 2 (ADAR2), in cultured astrocytes and astrocytes freshly isolated from transgenic mice tagged with an astrocyte-specific marker. In contrast, neurones isolated from transgenic mice tagged with a neurone-specific marker and exposed to fluoxetine showed an increase in gene expression of glutamate receptor, ionotropic kainate 4 (GluK4) and 5-hydroxytryptamine receptor 2C (5-HT2CR). In a mouse model of anhedonia, the downregulation of 5-HT2BR, cPLA2α, ADAR2 and GluK4 but not GluK2 and 5-HT2CR was detected.

Objective

To investigate the effects of chronic mild stress (CMS) and/or fluoxetine treatment on gene expression of 5-HT2BR, 5-HT2CR, cPLA2α, ADAR2, GluK2 and GluK4 specifically in astrocytes and neurones.

Methods

Transgenic mice tagged with either astrocyte- or neurone-specific markers were exposed to the CMS. Real-time PCR was applied to determine expression of messenger RNA (mRNA).

Results

We found that (i) mRNAs of the 5-HT2BR and cPLA2α in astrocytes and GluK4 in neurones were significantly reduced in mice that became anhedonic; the mRNA levels were restored by fluoxetine treatment; (ii) ADAR2 in astrocytes was decreased by the CMS but showed no response to fluoxetine in anhedonic animals; (iii) neither GluK2 expression in astrocytes nor 5-HT2CR expression in neurones were affected in anhedonic animals, although expression of 5-HT2CR mRNA was upregulated by fluoxetine.

Conclusions

Our results indicate that the effects of chronic treatment with fluoxetine are not only dependent on the cell type studied but also on the development of anhedonia. This suggests that fluoxetine may affect major depression (MD) patients and healthy people in a different manner.

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Abbreviations

5-HT2B 5-HT2C :

5-Hydroxytryptamine receptors 2B and 2C

ADAR:

Adenosine deaminases acting on RNA

APV:

2-Amino-5-phosphonovalerate

BSA:

Bovine serum albumin

CMS:

Chronic mild stress

cPLA2α :

Cytosolic phospholipase 2α

DNQX:

6,7-Dinitroquinoxaline-2,3-dione

EGF:

Epidermal growth factor

ERK:

Extracellular signal-regulated kinase

FACS:

Fluorescence-activated cell sorting

Fgfr3:

Fibroblast growth factor receptor 3

GABA:

γ-Aminobutyric acid

Gabra-1:

Gamma-aminobutyric acid receptor subunit α-1

GFAP:

Glial fibrillary acidic protein

GFP:

Green fluorescent protein promoter

Glt-1:

Glutamate transporter 1

GluK2 and GluK4:

Glutamate receptor, ionotropic kainate 2 and 4

KCC2:

Potassium-chloride transporter member 5

MEM:

Minimum essential medium

Mag:

Myelin-associated glycoprotein

Mbp:

Myelin basic protein

MD:

Major depression

Mog:

Myelin-oligodendrocyte glycoprotein

mRNA:

Messenger ribonucleic acid

PCR:

Polymerase chain reaction

PBS:

Phosphate-buffered saline

SSRI:

Serotonin-specific reuptake inhibitor

PI:

Propidium iodide

PIPES:

Piperazine-N,N’-bis(2-ethanesulfonic acid)

Snap25:

Synaptosomal-associated protein 25

TBP:

TATAA-box binding protein

Thy1:

Thymocyte antigen 1

YFP:

Yellow fluorescent protein

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Acknowledgments

This study was supported by grant nos. 31000479 to BL and 31171036 to LP from the National Natural Science Foundation of China. AV was supported in part by the grant (agreement from August 27 2013, No. 02.В.49.21.0003) between The Ministry of Education and Science of the Russian Federation and Lobachevsky State University of Nizhny Novgorod and by the grant of the Russian Scientific Foundation No. 14-15-00633.

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    Authors and Affiliations

    1. Laboratory of Brain Metabolic Diseases, Institute of Metabolic Disease Research and Drug Development, China Medical University, Shenyang, People’s Republic of China

      Lu Dong, Baoman Li & Liang Peng

    2. Faculty of Life Science, The University of Manchester, Manchester, UK

      Alexei Verkhratsky

    3. Achucarro Center for Neuroscience, IKERBASQUE, Basque Foundation for Science, 48011, Bilbao, Spain

      Alexei Verkhratsky

    4. University of Nizhny Novgorod, Nizhny, Novgorod, 603022, Russia

      Alexei Verkhratsky

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    1. Lu Dong
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    2. Baoman Li
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    4. Liang Peng
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    Correspondence to Liang Peng.

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    Lu Dong and Baoman Li contributed equally to this work.

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    Dong, L., Li, B., Verkhratsky, A. et al. Cell type-specific in vivo expression of genes encoding signalling molecules in the brain in response to chronic mild stress and chronic treatment with fluoxetine. Psychopharmacology 232, 2827–2835 (2015). https://doi.org/10.1007/s00213-015-3921-2

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