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Sympathetic neuron–associated macrophages contribute to obesity by importing and metabolizing norepinephrine

Nature Medicine volume 23, pages 1309–1318 (2017)Cite this article

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Abstract

The cellular mechanism(s) linking macrophages to norepinephrine (NE)-mediated regulation of thermogenesis have been a topic of debate. Here we identify sympathetic neuron–associated macrophages (SAMs) as a population of cells that mediate clearance of NE via expression of solute carrier family 6 member 2 (SLC6A2), an NE transporter, and monoamine oxidase A (MAOA), a degradation enzyme. Optogenetic activation of the sympathetic nervous system (SNS) upregulates NE uptake by SAMs and shifts the SAM profile to a more proinflammatory state. NE uptake by SAMs is prevented by genetic deletion of Slc6a2 or inhibition of the encoded transporter. We also observed an increased proportion of SAMs in the SNS of two mouse models of obesity. Genetic ablation of Slc6a2 in SAMs increases brown adipose tissue (BAT) content, causes browning of white fat, increases thermogenesis, and leads to substantial and sustained weight loss in obese mice. We further show that this pathway is conserved, as human sympathetic ganglia also contain SAMs expressing the analogous molecular machinery for NE clearance, which thus constitutes a potential target for obesity treatment.

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Figure 1: Sympathetic neuron–associated Cx3cr1-GFP+ cells exhibit differentiated morphology for specific association with SNS neurons.
Figure 2: SAMs highly express macrophage-associated markers and possess the machinery for uptake and degradation of norepinephrine.
Figure 3: SAMs import and metabolize norepinephrine via SLC6A2 and MAOA, respectively, to regulate extracellular norepinephrine availability.
Figure 4: Obesity-induced accumulation of SAMs.
Figure 5: Loss of Slc6a2 function in SAMs rescues the thermogenic capacity of ob/ob mice.
Figure 6: SAMs in the human sympathetic nervous system.

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Acknowledgements

We would like to thank the Unit for Imaging and Cytometry at the Instituto Gulbenkian de Ciência (IGC) for assistance with flow cytometry, cell sorting, and multiphoton microscopy. We also want to thank the Antibody Service at the IGC for the antibodies produced in house and the Histopathology facility at the IGC for tissue processing and histological assessment. This work was supported by the Fundação para a Ciéncia e Tecnologia (FCT), the European Molecular Biology Organization (EMBO), the Human Frontier Science Program (HFSP), Maratona da Saúde, and the US National Institutes of Health (NIH). R.M.P. was supported by FCT (SFRH/BD/88454/2012), J.S.S. was supported by the American Heart Association (16PRE30980030) and a training grant (T32DK007541), B.A.A. was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and N.M.-S. was supported by Xunta de Galicia (ED481B 2016/168-0). We thank M. Aouadi for helpful discussions.

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Author notes

  1. Roksana M Pirzgalska and Elsa Seixas: These authors contributed equally to this work.

Authors and Affiliations

  1. The Howard Hughes Medical Institute (HHMI) and Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras, Portugal

    Roksana M Pirzgalska, Elsa Seixas, Noelia Martínez Sánchez, Inês Mahú, Raquel Mendes, Vitka Gres, Nadiya Kubasova, Imogen Morris, Bernardo A Arús, Chelsea M Larabee, Miguel Vasques, Sathyavathy Anandan & Ana I Domingos

  2. Department of Cellular and Molecular Medicine, University of California, La Jolla, San Diego, California, USA

    Jason S Seidman, Verena M Link, Nathanael J Spann & Christopher K Glass

  3. Department II, Faculty of Biology, Ludwig-Maximilians Universität München, Planegg-Martinsried, Germany

    Verena M Link

  4. Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil

    Bernardo A Arús

  5. Department of Endocrinology, Curry Cabral Hospital, Centro Hospitalar de Lisboa Central, Lisbon, Portugal

    Miguel Vasques

  6. Department of Pathology, Centro Hospitalar Lisboa Norte, Hospital de Santa Maria, EPE, Lisbon, Portugal

    Francisco Tortosa

  7. Electron Microscopy Unit, Instituto Gulbenkian de Ciência, Oeiras, Portugal

    Ana L Sousa & Erin Tranfield

  8. Vanderbilt Kennedy Center for Research on Human Development, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Maureen K Hahn

  9. Division of Immunology, Transplantation and Infectious Diseases, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Scientific Institute, Milan, Italy

    Matteo Iannacone

  10. The Howard Hughes Medical Institute (HHMI), New York, New York, USA

    Ana I Domingos

Authors
  1. Roksana M Pirzgalska
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Contributions

A.I.D. conceptualized the study. R.M.P. performed two-photon and confocal microscopy. E.S. and R.M.P. performed flow cytometry. J.S.S. and R.M.P. performed low-input RNA-seq. V.M.L., J.S.S., and R.M.P. analyzed the RNA-seq data. M.I., A.L.S., S.A., and E.T. performed electron microscopy. E.S., R.M.P., N.M.S., I. Mahú, B.A.A., and C.M.L. performed functional tests. N.K., I. Morris, R.M., and V.G. performed related mouse husbandry and genotyping. F.T. and M.V. processed human ganglia. M.K.H. provided the Slc6a2−/− mice. N.J.S. developed the low-input RNA-seq protocols. A.I.D., C.K.G., and R.M.P. wrote the original draft of the manuscript. A.I.D., C.K.G., R.M.P., and C.M.L. reviewed and edited the final version of the manuscript.

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Correspondence to Ana I Domingos.

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Supplementary Figures 1–12 and Supplementary Table 1 (PDF 20527 kb)

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In vivo visualization of SAMs in the neuro-adipose connection.

Intra-vital multi-photon visualization of a neuro-adipose connection in the inguinal fat pad of a live Cx3cr1GFP/+ mouse; LipidTOX (blue) labels adipocytes. Images are representative of 3 similar experiments. (MPG 192 kb)

In vivo visualization of ATMs in the subcutaneous adipose tissue

Intra-vital multi-photon visualization of the inguinal fat pad of a live Cx3cr1GFP/+ mouse; LipidTOX (blue) labels adipocytes. Images are representative of 3 similar experiments. (MPG 56 kb)

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Pirzgalska, R., Seixas, E., Seidman, J. et al. Sympathetic neuron–associated macrophages contribute to obesity by importing and metabolizing norepinephrine. Nat Med 23, 1309–1318 (2017). https://doi.org/10.1038/nm.4422

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