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Measuring neuron-regulated immune cell physiology via the alpha-2 adrenergic receptor in an ex vivo murine spleen model

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Abstract

The communication between the nervous and immune systems plays a crucial role in regulating immune cell function and inflammatory responses. Sympathetic neurons, which innervate the spleen, have been implicated in modulating immune cell activity. The neurotransmitter norepinephrine (NE), released by sympathetic neurons, influences immune cell responses by binding to adrenergic receptors on their surface. The alpha-2 adrenergic receptor (α2AR), expressed predominantly on sympathetic neurons, has received attention due to its autoreceptor function and ability to modulate NE release. In this study, we used fast-scan cyclic voltammetry (FSCV) to provide the first subsecond measurements of NE released in the white pulp region of the spleen and validated it with yohimbine, a known antagonist of α2AR. For further application of FSCV in neuroimmunology, we investigated the extent to which subsecond NE from sympathetic neurons is important for immune cell physiology and cytokine production, focusing on tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), and interleukin-6 (IL-6). Our findings provide insights into the regulatory mechanisms underlying sympathetic-immune interactions and show the significance of using FSCV, a traditional neurochemistry technique, to study these neuroimmune mechanisms.

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The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author. The data will be made available from the corresponding author on reasonable request.

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Acknowledgements

The authors of this work would like to acknowledge the assistance of the Live Microscopy Core in the Department of Pharmacology & Systems Physiology at the University of Cincinnati College of Medicine. Abstract image and figure one were created with the help of Biorender.com.

Funding

The research reported in this manuscript was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number R01AI151552. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The authors would also like to thank the Alfred P. Sloan Foundation (Grant #FG-2022-18400) for support of this work. The authors would also like to acknowledge the NIH T32 (NS007453) for funding K.C.N.C. for this work.

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  1. Department of Chemistry, University of Cincinnati, 312 College Dr. 404 Crosley Tower, Cincinnati, OH, 45221-0172, USA

    Alexandra K. Brooke, Daniel P. Murrow, Kaejaren C. N. Caldwell, Colby E. Witt & Ashley E. Ross

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  1. Alexandra K. Brooke
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AKB and AER contributed to the study concept and design. Material preparation, data collection and analysis were performed by AKB, DPM, KCNC, and CEW. The first draft of the manuscript was written by AKB. AKB, CEW, and AER contributed on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ashley E. Ross.

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Brooke, A.K., Murrow, D.P., Caldwell, K.C.N. et al. Measuring neuron-regulated immune cell physiology via the alpha-2 adrenergic receptor in an ex vivo murine spleen model. Cell. Mol. Life Sci. 80, 354 (2023). https://doi.org/10.1007/s00018-023-05012-2

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