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Identification of a unique TGF-β–dependent molecular and functional signature in microglia

Nature Neuroscience 17, 131–143 (2014) | Download Citation

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  • A Corrigendum to this article was published on 26 August 2014

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Abstract

Microglia are myeloid cells of the CNS that participate both in normal CNS function and in disease. We investigated the molecular signature of microglia and identified 239 genes and 8 microRNAs that were uniquely or highly expressed in microglia versus myeloid and other immune cells. Of the 239 genes, 106 were enriched in microglia as compared with astrocytes, oligodendrocytes and neurons. This microglia signature was not observed in microglial lines or in monocytes recruited to the CNS, and was also observed in human microglia. We found that TGF-β was required for the in vitro development of microglia that express the microglial molecular signature characteristic of adult microglia and that microglia were absent in the CNS of TGF-β1–deficient mice. Our results identify a unique microglial signature that is dependent on TGF-β signaling and provide insights into microglial biology and the possibility of targeting microglia for the treatment of CNS disease.

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  • 19 December 2013

    In the version of this article initially published, the x-axis labels for the sets of graphs in Figure 2f corresponding to astrocyte, oligodendrocyte and neuron molecules consisted of six items, even though there were only five bars. “Red pulp macrophages” was included in error. Also, the Cleveland Clinic affiliation gave the section as the Department of Immunology; the correct affiliation is Neuroinflammation Research Center. The errors have been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank D. Julius (University of California, San Francisco) for providing polyclonal antibody to P2ry12 and N. Kassam for support in antibody generation, A. Krichevsky and A. Wong Hon-Kit (Brigham and Women's Hospital, Harvard Medical School) for providing neurons, and L. Spangler for technical assistance for oligodendrocyte isolation. We thank D. Kozoriz for the FACS sorting. This work was supported by US National Institutes grant AG027437, US National Institutes Transformative Grant AG-043975, a grant from the Amyotrophic Lateral Sclerosis Association (1V78RI, ALSA 2087), a grant from Department of Defense ALS Research Program (AL120029), a Thome Foundation AMD grant, and philanthropic support. We thank Prize4Life for providing SOD1 mice.

Author information

Affiliations

  1. Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Oleg Butovsky
    • , Ron Cialic
    • , Amanda J Lanser
    • , Galina Gabriely
    • , Thomas Koeglsperger
    • , Ben Dake
    • , Pauline M Wu
    • , Camille E Doykan
    • , Zain Fanek
    •  & Howard L Weiner

  2. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.

    • Mark P Jedrychowski
    •  & Steven P Gygi

  3. Neuroimmunology Unit, Montréal Neurological Institute, McGill University, Montréal, Québec, Canada.

    • Craig S Moore
    •  & Jack P Antel

  4. Neuroinflammation Research Center, Cleveland Clinic, Cleveland, Ohio, USA.

    • LiPing Liu
    •  & Richard M Ransohoff

  5. Brain Science Institute and Department of Neurology, Johns Hopkins University, Baltimore, Maryland, USA.

    • Zhuoxun Chen
    •  & Jeffrey D Rothstein

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Contributions

O.B. and H.L.W. conceived the study, designed the experiments and wrote the paper. O.B., A.J.L., G.G., T.K., B.D., R.C., P.M.W., C.E.D. and Z.F. performed experiments. M.P.J. and S.P.G. performed mass spectrometry experiments. C.S.M. and J.P.A. performed human microglia studies. Z.C., J.D.R., L.L. and R.M.R. performed CNS cell isolation studies. All authors discussed the results and conclusions and reviewed the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Oleg Butovsky or Howard L Weiner.

Integrated supplementary information

Supplementary figures

  1. 1.

    Identification of protein signature in microglia and Ly6C monocyte subsets.

  2. 2.

    Top biological functions in microglia.

  3. 3.

    Nervous system development and function in microglia.

  4. 4.

    Top microglial interactions by protein function.

  5. 5.

    Canonical pathways in microglia and monocytes.

  6. 6.

    TGF-β pathway in microglia

  7. 7.

    miRNA profile in microglia vs. astrocytes, oligodendrocytes and neurons.

  8. 8.

    Specificity of P2ry12 and FCRLS antibodies in resident microglia.

  9. 9.

    Microglia signature during development.

  10. 10.

    M0, M1 and M2 microglial phenotypes as measured by MG400 chip.

  11. 11.

    Microglia loss in CNS-TGFβ1−/− mice.

  12. 12.

    Macrophages, dendritic cells and Langerhans cells are not affected in peripheral organs of CNS-TGFβ1−/− mice.

  13. 13.

    Loss of microglia during development in CNS-TGF-β1 deficient mice

  14. 14.

    Microglia loss in the spinal cord of CNS-TGFβ1−/− mice.

  15. 15.

    TGF-β pathway and downstream microglial molecules are suppressed in CNS-TGFβ1−/− mice.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–15 and Supplementary Tables 1–4

Excel files

  1. 1.

    Supplementary Table 2

    Top unique microglial genes and shared genes between microglia, neurons, astrocytes and oligodendrocytes. List of 152 identified genes which are enriched in adult microglia and not in adult astrocytes, oligodendrocytes and neurons.

Videos

  1. 1.

    Rotorod performance of CNS-TGFβ1−/− mice

    IL2TGFβ1-Tg-TGF-β1−/− (left) and IL2TGFβ1-Tg-TGF-β1+/− (right) mice at the age of 140 days of age.

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DOI

https://doi.org/10.1038/nn.3599

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