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Hypothalamic Proteomic Analysis Reveals Dysregulation of Glutamate Balance and Energy Metabolism in a Mouse Model of Chronic Mild Stress-Induced Depression

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Abstract

Hypothalamus–pituitary–adrenal (HPA) axis hyperactivity is observed in many patients suffering from depression. However, the mechanism underlying the dysfunction of the HPA axis is not well understood. Moreover, dysfunction of the hypothalamus, the key brain region of the HPA axis, has not been well-explored. The aim of our study was to examine possible alterations in hypothalamus protein expression in a model of depression using proteomic analysis. In order to achieve this aim, mice were exposed to chronic unpredictable mild stress (CUMS), as the paradigm results in hyperactivity of the HPA axis. Differential protein expression between the hypothalamic proteomes of CUMS and control mice was then assessed through two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry. Thirty-seven proteins with a threshold of a 1.5-fold change and a p value ≤0.05 were identified as being differentially expressed between CUMS and control mice, and were quantified for bioinformatics analysis. Glycometabolism, citrate cycle (TCA cycle) and oxidation respiratory chain were found to have changed significantly. Glial fibrillary acidic protein and glutamine synthetase were further validated by Western Blot. Our results demonstrated that CUMS mice exhibited a dramatic protein change both in glutamate metabolism and energy mobilization, which may shed some light on the role of the hypothalamus in the pathology of stress-induced depression.

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Abbreviations

2-DE:

Two-dimensional electrophoresis

ACTH:

Adrenocorticotropic hormone

CON:

Control

CRF:

Corticotrophin-releasing factor

CUMS:

Chronic unpredictable mild stress

DAVID:

Database for annotation, visualization and integrated discovery

FST:

Forced swimming test

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GFAP:

Glial fibrillary acidic protein

GO:

Gene ontology

GR:

Glucocorticoid receptor

GS:

Glutamine synthetase

HPA:

Hypothalamus–pituitary–adrenal

IEF:

Isoelectric focusing

KEGG:

Kyoto Encyclopedia of Genes and Genomes

MALDI-TOF–MS/MS:

Matrix-assisted laser desorption ionization-time of flight–tandem mass spectrometry

MW:

Molecular weight

OD:

Optical density

OFT:

Open field test

pI:

Isoelectric point

SPSS:

Statistical Package of Social Science

SPT:

Sucrose preference test

TCA:

Tricarboxylic acid

UniProt:

Universal protein resource

MDD:

Major depressive disorder

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Acknowledgments

We thank Dr. ChenruiHou (Shanghai Institute for Biological Sciences, Chinese Academy of Sciences) for his assistance with the mass spectrometric analysis. We would like to thank Editage (http://www.editage.cn/) for English language editing and proofreading this manuscript. This work was supported by grants from the National Basic Research Program of China (“973 program”, Grant No. 2009CB918300). The Health Bureau medical scientific research projects of Chongqing (Grant No. 20142022).

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    Authors and Affiliations

    1. Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing, China

      Chenglong Rao, Haiyang Shi, Chanjuan Zhou, Jianyong Tang & Peng Xie

    2. Chongqing Key Laboratory of Neurobiology, Chongqing, China

      Chenglong Rao, Haiyang Shi, Chanjuan Zhou, Dan Zhu, Mingjun Zhao, Ziye Wang, Yongtao Yang, Jin Chen, Li Liao, Jianyong Tang, You Wu, Jian Zhou, Ke Cheng & Peng Xie

    3. Institute of Neuroscience and the Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing, China

      Chenglong Rao, Haiyang Shi, Chanjuan Zhou, Dan Zhu, Mingjun Zhao, Ziye Wang, Yongtao Yang, Jin Chen, Li Liao, Jianyong Tang, You Wu, Jian Zhou, Ke Cheng & Peng Xie

    4. Department of Neurology, The First Affiliated Hospital, Chongqing Medical University, No. 1 Yixue Road, Yuzhong District, Chongqing, 400016, China

      Dan Zhu, You Wu, Ke Cheng & Peng Xie

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    1. Chenglong Rao
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    Correspondence to Peng Xie.

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    Chenglong Rao, Haiyang Shi, Chanjuan Zhou and Dan Zhu have contributed equally to this work.

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    Supplemental Fig. 1

    Time schedule of procedures used in the present study. After 7 days of acclimatization, mice were trained to adapt 1 % (w/v) sucrose solution. Then a baseline of sucrose preference was gotten for group dividing, CUMS procedures were performed for 28 days. The sucrose preference test (SPT) and body weights were conducted weekly throughout the CUMS period. Open field test (OFT) and forced swimming test (FST) were executed respectively on day 29 and day 30. Tissue separated at the end of the schedule (TIFF 106 kb)

    Supplemental Fig. 2

    The sketch map of the molecular dysregulations supporting stress-induced depressive-like behaviors. Dysregulation of glutamatergic synapses may result in the accumulation of glutamate in the synaptic cleft, exciting the HPA axis (TIFF 353 kb)

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    Rao, C., Shi, H., Zhou, C. et al. Hypothalamic Proteomic Analysis Reveals Dysregulation of Glutamate Balance and Energy Metabolism in a Mouse Model of Chronic Mild Stress-Induced Depression. Neurochem Res 41, 2443–2456 (2016). https://doi.org/10.1007/s11064-016-1957-2

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