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Downregulation of CRTC1 Is Involved in CUMS-Induced Depression-Like Behavior in the Hippocampus and Its RNA Sequencing Analysis

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Abstract

Chronic stress is an important risk factor for mood disorders including depression. The decreased level of CREB (cAMP-responsive element binding)-regulated transcription coactivator 1 (CRTC1) expression in hippocampus may be involved in depression-like behavior in some stress-induced depression models. But the mechanism of CRTC1 in mediating depression-like behavior remains unknown. In this study, chronic unpredictable mild stress (CUMS)-treated mice showed depression-like behavior accompanied by the downregulation of CRTC1 in the hippocampus. Adeno-associated virus (AAV)-CRTC1-mediated overexpression of CRTC1 in the hippocampus by stereotactic brain injection could significantly prevent depression-like behavior in CUMS-treated mice. The above data reveal that the downregulation of hippocampal CRTC1 expression participates in CUMS-induced depression-like behavior. In order to explore the key targets regulated by CRTC1, AAV-mediated CRTC1 short hairpin (shRNA) was constructed to achieve knockdown of CRTC1 in the hippocampus, and then the hippocampi were collected for RNA-sequencing (RNA-seq). The RNA-seq data show that upregulated genes were enriched in stress and immune system-associated GO terms and pathways such as response to stress and external stimulus and regulation of immune response and that downregulated genes were enriched in neural activity such as synaptic transmission and cognitive behavior. We further provided RT-qPCR data that the inflammation-related factors including Gpr84, Tlr2, Lyz2, and Icam1 were significantly upregulated in the hippocampus of both CUMS- and CRTC1 shRNA-induced models, some of them were also validated in protein levels by Western blotting. We propose a hypothesis that CUMS induces downregulation of CRTC1, which might lead to depression-like behavior via neuroinflammation pathway. This study provides new explanation for the inflammatory hypothesis of depression and some clues for exploring the molecular mechanism of CRTC1 regulation.

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Data Availability

The RNA-seq raw data after Crtc1 knockdown are included as supplementary files in this publication.

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Acknowledgements

We thank for the technical support by the Core Facilities, Ningbo University School of Medicine.

Funding

This research was supported by the National Natural Science Foundation of China [grant numbers 81171036, 81371224], by the Natural Science Foundation of Zhejiang Province [grant number LY19H090003] and by the Natural Science Foundation of Ningbo [grant number 2018A610288]. This project was also sponsored by K.C. Wong Magna Fund in Ningbo University.

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  1. Dezhu Li, Qi Liao and Yang Tao contributed equally to this work.

Authors and Affiliations

  1. The Affiliated Hospital of Medical School, Ningbo University, Ningbo, 315211, China

    Dezhu Li, Qi Liao, Yang Tao, Saiqi Ni, Chuang Wang, Dingli Xu, Xiaowei Chen, Wei Cui & Junfang Zhang

  2. Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, Ningbo, 315211, China

    Saiqi Ni, Chuang Wang, Xiaowei Chen, Wei Cui & Junfang Zhang

  3. Ningbo Kangning Hospital, Ningbo, 315210, China

    Dongsheng Zhou & Xingxing Li

  4. Department of Human Anatomy, School of Basic Medical Sciences, Capital Medical University, Beijing, 100069, China

    Xinchun Jin

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Contributions

FZ designed and supervised the study. DZL performed the stereotaxic delivery, behavioral tests, and RT-qPCR with the help of CW and XWC. YT performed RNA-seq analysis with the help of QL and DSZ. SQN performed virus verification and WB with the help of XXL. DLX and XCJ contributed to the data analysis. JFZ, DZL, and QL wrote the manuscript, JFZ secured funding for the project. All authors discussed the results and approved the final manuscript.

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Correspondence to Junfang Zhang.

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All animal experiments were carried out in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals (8th edition, National Research Council) and approved by the Institutional Animal Care and Use Committee of the Medical School of Ningbo University (SYXK(ZHE)2013–0191).

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Li, D., Liao, Q., Tao, Y. et al. Downregulation of CRTC1 Is Involved in CUMS-Induced Depression-Like Behavior in the Hippocampus and Its RNA Sequencing Analysis. Mol Neurobiol 59, 4405–4418 (2022). https://doi.org/10.1007/s12035-022-02787-6

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