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Novel Hexb-based tools for studying microglia in the CNS

Nature Immunology volume 21, pages 802–815 (2020)Cite this article

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An Author Correction to this article was published on 11 August 2020

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Abstract

Microglia and central nervous system (CNS)-associated macrophages (CAMs), such as perivascular and meningeal macrophages, are implicated in virtually all diseases of the CNS. However, little is known about their cell-type-specific roles in the absence of suitable tools that would allow for functional discrimination between the ontogenetically closely related microglia and CAMs. To develop a new microglia gene targeting model, we first applied massively parallel single-cell analyses to compare microglia and CAM signatures during homeostasis and disease and identified hexosaminidase subunit beta (Hexb) as a stably expressed microglia core gene, whereas other microglia core genes were substantially downregulated during pathologies. Next, we generated HexbtdTomato mice to stably monitor microglia behavior in vivo. Finally, the Hexb locus was employed for tamoxifen-inducible Cre-mediated gene manipulation in microglia and for fate mapping of microglia but not CAMs. In sum, we provide valuable new genetic tools to specifically study microglia functions in the CNS.

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Fig. 1: scRNA-seq during de- and remyelination and FNX reveals Hexb as a stably expressed microglia core gene.
Fig. 2: Steady high expression of Hexb in microglia but not CAMs during homeostasis and autoimmune neuroinflammatory conditions.
Fig. 3: Generation and characterization of HexbtdTomato mice.
Fig. 4: Continuous detection of HexbtdT microglia during neurodegeneration and autoimmune demyelination.
Fig. 5: In vivo dynamics of HexbtdT microglia.
Fig. 6: Generation and comparative analysis of HexbCreERT2 mice.
Fig. 7: Fate mapping following CNS macrophage depletion using HexbCreERT2/CreERT2 R26yfp/yfp mice.
Fig. 8: Effects of Csf1r deletion on microglia but not CAMs.

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Data availability

Raw data for new scRNA-seq or bulk RNA-seq have been deposited in the Gene Expression Omnibus, and are available at the following accession numbers: GSE148405 (scRNA-seq) and GSE148413 (bulk RNA-seq). All other data are published previously or are available from the corresponding authors on reasonable request.

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Acknowledgements

We thank E. Barleon for excellent technical assistance and the Center for Animal Resource and Development (CARD, Kumamoto University) for providing research material. T. Masuda was supported by the KANAE Foundation for the Promotion of Medical Science and the Japan Society for the Promotion of Science (JSPS) as the JSPS Postdoctoral Fellow for Research Abroad. M.P. was supported by the Sobek Foundation, the Ernst-Jung Foundation, the German Research Foundation (DFG) (grant nos. SFB 992, SFB1160, SFB/TRR167, Reinhart-Koselleck-Grant and the Gottfried Wilhelm Leibniz-Prize) and the Ministry of Science, Research and Arts, Baden-Wuerttemberg (Sonderlinie ‘Neuroinflammation’). This study was supported by the DFG under Germany’s Excellence Strategy (grant no. CIBSS—EXC-2189, Project ID390939984). J.P. was supported by the DFG (grant nos. SFB/TRR167 B05 and B07), and the UK DRI Momentum Award. C.B. was supported by the DFG (grant no. SFB/TRR167 B05).

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Author notes

  1. These authors contributed equally: Takahiro Masuda, Lukas Amann.

Authors and Affiliations

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

    Takahiro Masuda, Lukas Amann, Roman Sankowski, Ori Staszewski, Marta Joana Costa Jordão, Katrin Kierdorf, Klaus-Peter Knobeloch & Marco Prinz

  2. Department of Life Innovation, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

    Takahiro Masuda

  3. Faculty of Biology, University of Freiburg, Freiburg, Germany

    Lukas Amann

  4. Berta-Ottenstein-Programme for Clinician Scientists, Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Roman Sankowski & Ori Staszewski

  5. Department of Neuroanatomy, Institute of Anatomy and Cell Biology, Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Maximilian Lenz & Andreas Vlachos

  6. Department of Neurology, Medical Center—University of Freiburg, Freiburg, Germany

    Paolo d´Errico & Melanie Meyer-Luehmann

  7. Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Paolo d´Errico & Melanie Meyer-Luehmann

  8. Institute of Neuronal Cell Biology, Technical University of Munich, Munich, Germany

    Nicolas Snaidero & Thomas Misgeld

  9. Department of Neuropsychiatry and Laboratory of Molecular Psychiatry, Charité—Universitätsmedizin Berlin, Berlin, Germany

    Chotima Böttcher & Josef Priller

  10. Center for Basics in NeuroModulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Katrin Kierdorf, Andreas Vlachos, Melanie Meyer-Luehmann & Marco Prinz

  11. CIBSS—Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany

    Katrin Kierdorf

  12. Department of Immunology, Weizmann Institute of Science, Rehovot, Israel

    Steffen Jung

  13. DZNE and BIH, Berlin, Germany

    Josef Priller

  14. University of Edinburgh and UK DRI, Edinburgh, UK

    Josef Priller

  15. German Center for Neurodegenerative Diseases, Munich, Germany

    Thomas Misgeld

  16. Munich Cluster for Systems Neurology, Munich, Germany

    Thomas Misgeld

  17. Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany

    Marco Prinz

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  1. Takahiro Masuda
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Contributions

T. Masuda, L.A., P.E., M.L., N.S., T. Misgeld, R.S., O.S., M.J.C.J., C.B., K.K., D.G., A.V. and K.P.K. conducted experiments and analyzed the data. M.P., K.P.K., S.J., M.M.L. and J.P. analyzed the data, contributed to the in vivo studies and provided mice or reagents. T. Masuda and M.P. supervised the project and wrote the manuscript, J.P. edited the manuscript.

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Correspondence to Takahiro Masuda or Marco Prinz.

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Masuda, T., Amann, L., Sankowski, R. et al. Novel Hexb-based tools for studying microglia in the CNS. Nat Immunol 21, 802–815 (2020). https://doi.org/10.1038/s41590-020-0707-4

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