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Chemogenetic manipulation of CX3CR1+ cells transiently induces hypolocomotion independent of microglia

Molecular Psychiatry volume 28, pages 2857–2871 (2023)Cite this article

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Abstract

Chemogenetic approaches using Designer Receptors Exclusively Activated by Designer Drugs (DREADD, a family of engineered GPCRs) were recently employed in microglia. Here, we used Cx3cr1CreER/+:R26hM4Di/+ mice to express Gi-DREADD (hM4Di) on CX3CR1+ cells, comprising microglia and some peripheral immune cells, and found that activation of hM4Di on long-lived CX3CR1+ cells induced hypolocomotion. Unexpectedly, Gi-DREADD-induced hypolocomotion was preserved when microglia were depleted. Consistently, specific activation of microglial hM4Di cannot induce hypolocomotion in Tmem119CreER/+:R26hM4Di/+ mice. Flow cytometric and histological analysis showed hM4Di expression in peripheral immune cells, which may be responsible for the hypolocomotion. Nevertheless, depletion of splenic macrophages, hepatic macrophages, or CD4+ T cells did not affect Gi-DREADD-induced hypolocomotion. Our study demonstrates that rigorous data analysis and interpretation are needed when using Cx3cr1CreER/+ mouse line to manipulate microglia.

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Fig. 1: Activation of Gi-DREADD in CX3CR1+ cells induces hypolocomotion in mice.
Fig. 2: Gi-DREADD-induced hypolocomotion is independent of microglia.
Fig. 3: CX3CR1 is expressed in peripheral cells and Cx3cr1-controlled tdTomato (tdT) reporter labels circulating leukocytes.
Fig. 4: Cx3cr1-controlled tdTomato (tdT) reporter is expressed in peripheral organs.
Fig. 5: Gi-DREADD is expressed in peripheral immune cells.
Fig. 6: Resident macrophages or CD4+ T cells are not required for Gi-DREADD-induced hypolocomotion.

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Acknowledgements

We thank Dr. Vanda A. Lennon (Mayo Clinic), Dr. Doo-Sup Choi (Mayo Clinic) and Dr. Bryan Roth (University of North Carolina) for thoughtful comments. We also thank members of the Wu lab for insightful discussions.

Funding

This work was supported by the following grants from the National Institutes of Health: R35NS132326 (L-JW), R01ES033892 (JRR, L-JW), and K99NS126417 (ADU).

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  1. These authors contributed equally: Shunyi Zhao, Jiaying Zheng.

Authors and Affiliations

  1. Department of Neurology, Mayo Clinic, Rochester, MN, USA

    Shunyi Zhao, Jiaying Zheng, Lingxiao Wang, Anthony D. Umpierre, Sebastian Parusel, Manling Xie, Aastha Dheer, Aaron J. Johnson & Long-Jun Wu

  2. Mayo Clinic Graduate School of Biomedical Sciences, Rochester, MN, USA

    Shunyi Zhao, Jiaying Zheng & Lingxiao Wang

  3. Department of Immunology, Mayo Clinic, Rochester, MN, USA

    Katayoun Ayasoufi, Aaron J. Johnson & Long-Jun Wu

  4. Department of Molecular Medicine, Mayo Clinic, Rochester, MN, USA

    Aaron J. Johnson

  5. Department of Environmental Health Science, Robert Stempel College of Public Health and Social Work, Florida International University, Miami, FL, USA

    Jason R. Richardson

  6. Department of Neuroscience, Mayo Clinic, Jacksonville, FL, USA

    Long-Jun Wu

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Contributions

SZ, JZ, and L-JW designed experiments and wrote the manuscript. SZ and JZ performed most of the experiments. LW performed data mining and analyzed RNA sequencing data. ADU, SP, and MX performed some experiments. SZ and AD analyzed data. KA, AJJ, and JRR provided resources for specific experiments.

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Correspondence to Long-Jun Wu.

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Zhao, S., Zheng, J., Wang, L. et al. Chemogenetic manipulation of CX3CR1+ cells transiently induces hypolocomotion independent of microglia. Mol Psychiatry 28, 2857–2871 (2023). https://doi.org/10.1038/s41380-023-02128-6

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