Abstract

Microglia constitute a highly specialized network of tissue-resident immune cells that is important for the control of tissue homeostasis and the resolution of diseases of the CNS. Little is known about how their spatial distribution is established and maintained in vivo. Here we establish a new multicolor fluorescence fate mapping system to monitor microglial dynamics during steady state and disease. Our findings suggest that microglia establish a dense network with regional differences, and the high regional turnover rates found challenge the universal concept of microglial longevity. Microglial self-renewal under steady state conditions constitutes a stochastic process. During pathology this randomness shifts to selected clonal microglial expansion. In the resolution phase, excess disease-associated microglia are removed by a dual mechanism of cell egress and apoptosis to re-establish the stable microglial network. This study unravels the dynamic yet discrete self-organization of mature microglia in the healthy and diseased CNS.

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Acknowledgements

The authors thank H. Snippert (University Medical Center Utrecht) and H. Clevers (Hubrecht Institute) for the R26RConfetti mouse line, M. Follo and team at Lighthouse Fluorescence Technologies Core Facility, T. Blank for critical feedback, J. Bührer for technical support, and CEMT, University of Freiburg for excellent animal care. T.L.T. is supported by the German Research Foundation (DFG, TA1029/1-1) and Ministry of Science, Research and the Arts, Baden-Wuerttemberg (Research Seed Capital). M.P. is supported by the BMBF Competence Network of Multiple Sclerosis (KKNMS), the Sobek-Stiftung and the DFG (SFB992, SFB1140, Reinhart-Koselleck grant). F.F.-K., J.A.P., S.J., J.P. and M.P. are supported by the SFB/TRR167 “NeuroMac”.

Author information

Affiliations

  1. Institute of Neuropathology, Faculty of Medicine, University of Freiburg, Freiburg, Germany.

    • Tuan Leng Tay
    • , Jana Dautzenberg
    • , Moumita Datta
    • , Alberto Ardura-Fabregat
    • , Ori Staszewski
    •  & Marco Prinz
  2. Institute for Computer Science, University of Freiburg, Freiburg, Germany.

    • Dominic Mai
    •  & Olaf Ronneberger
  3. Department of Neuropsychiatry & Laboratory of Molecular Psychiatry, Charité – Universitätsmedizin Berlin, Berlin, Germany.

    • Francisco Fernández-Klett
    •  & Josef Priller
  4. Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Gen Lin
    •  & Lars M Steinmetz
  5. Max Planck Institute of Immunobiology and Epigenetics, Freiburg, Germany.

    • Sagar
    • , Anne Drougard
    • , J Andrew Pospisilik
    •  & Dominic Grün
  6. Center of Excellence for Fluorescent Bioanalytics, University of Regensburg, Regensburg, Germany.

    • Thomas Stempfl
  7. Advanced Light Microscopy Technology Platform, Max Delbrück Center for Molecular Medicine, Berlin, Germany.

    • Anca Margineanu
    •  & Anje Sporbert
  8. Department of Genetics, Stanford University School of Medicine, Stanford, California, USA.

    • Lars M Steinmetz
  9. Stanford Genome Technology Center, Palo Alto, California, USA.

    • Lars M Steinmetz
  10. Department of Immunology, Weizmann Institute of Science, Rehovot, Israel.

    • Steffen Jung
  11. Cluster of Excellence 'NeuroCure', German Center for Neurodegenerative Diseases (DZNE) and Berlin Institute of Health (BIH), Berlin, Germany.

    • Josef Priller
  12. BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany.

    • Marco Prinz

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Contributions

T.L.T. and M.P. conceived the study. T.L.T. established the methods. D.M., O.R. and T.L.T. designed the software and validated imaging data. T.L.T., J.D., F.F.-K., G.L., S., D.G., M.D., A.D., T.S., A.A.-F., O.S., A.M., A.S. and S.J. performed experiments and/or analyses. T.L.T., L.M.S., J.A.P., D.G., J.P., O.R. and M.P. provided supervision. T.L.T., D.M. and M.P. wrote the manuscript. All authors contributed to the editing of the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Tuan Leng Tay or Marco Prinz.

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    Supplementary Text and Figures

    Supplementary Figures 1–5 and Supplementary Tables 1–3.

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    Supplementary Methods Checklist

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Videos

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    Supplementary Video 1

    Thirty-minute time lapse recording of steady state GFP+ adult microglial cells in a CX3CR1GFP/wt mouse via a cranial window.

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    Supplementary Video 2

    Thirty-minute time lapse recording of a steady state RFP+ adult microglial cell in a Microfetti mouse via a cranial window.

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DOI

https://doi.org/10.1038/nn.4547

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