Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Loss of microglial EED impairs synapse density, learning, and memory

Molecular Psychiatry volume 27, pages 2999–3009 (2022)Cite this article

Subjects

Abstract

The embryonic ectoderm development (EED) is a core component of the polycomb-repressive complex 2 (PRC2) whose mutations are linked to neurodevelopmental abnormalities, intellectual disability, and neurodegeneration. Although EED has been extensively studied in neural stem cells and oligodendrocytes, its role in microglia is incompletely understood. Here, we show that microglial EED is essential for synaptic pruning during the postnatal stage of brain development. The absence of microglial EED at early postnatal stages resulted in reduced spines and impaired synapse density in the hippocampus at adulthood, accompanied by upregulated expression of phagocytosis-related genes in microglia. As a result, deletion of microglial Eed impaired hippocampus-dependent learning and memory in mice. These results suggest that microglial EED is critical for normal synaptic and cognitive functions during postnatal development.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Microglia-specific depletion of Eed affects microglia number and microglial morphology.
Fig. 2: Loss of microglial Eed leads to reductions in neuronal activity, dendritic spine density, and expression levels of PSD-95 and synaptophysin.
Fig. 3: Microglia-specific deletion of Eed impairs hippocampus-dependent learning and memory in mice.
Fig. 4: EED is required for normal synapse engulfment in vivo.
Fig. 5: Microglia-specific depletion of Eed alters the expression of genes associated with phagocytosis and engulfment.
Fig. 6: Deletion of Eed in microglia activates expression of prophagocytosis genes.

Similar content being viewed by others

Data availability

The microglia RNA-seq data have been deposited in the Genome Sequence Archive in the National Genomics Data Center, Beijing Institute of Genomics, Chinese Academy of Sciences with accession number CRA006251.

References

  1. Liu PP, Xu YJ, Dai SK, Du HZ, Wang YY, Li XG, et al. Polycomb protein EED regulates neuronal differentiation through targeting SOX11 in hippocampal dentate Gyrus. Stem Cell Rep. 2019;13:115–31.

    Article  CAS  Google Scholar 

  2. Ueda T, Nakata Y, Nagamachi A, Yamasaki N, Kanai A, Sera Y, et al. Propagation of trimethylated H3K27 regulated by polycomb protein EED is required for embryogenesis, hematopoietic maintenance, and tumor suppression. Proc Natl Acad Sci USA. 2016;113:10370–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Margueron R, Justin N, Ohno K, Sharpe ML, Son J, Drury WJ 3rd, et al. Role of the polycomb protein EED in the propagation of repressive histone marks. Nature. 2009;461:762–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ueda T, Sanada M, Matsui H, Yamasaki N, Honda ZI, Shih LY, et al. EED mutants impair polycomb repressive complex 2 in myelodysplastic syndrome and related neoplasms. Leukemia. 2012;26:2557–60.

    Article  CAS  PubMed  Google Scholar 

  5. Lessard J, Schumacher A, Thorsteinsdottir U, van Lohuizen M, Magnuson T, Sauvageau G. Functional antagonism of the Polycomb-Group genes eed and Bmi1 in hemopoietic cell proliferation. Genes Dev. 1999;13:2691–703.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Richie ER, Schumacher A, Angel JM, Holloway M, Rinchik EM, Magnuson T. The polycomb-group gene eed regulates thymocyte differentiation and suppresses the development of carcinogen-induced T-cell lymphomas. Oncogene. 2002;21:299–306.

    Article  CAS  PubMed  Google Scholar 

  7. Sauvageau M, Miller M, Lemieux S, Lessard J, Hebert J, Sauvageau G. Quantitative expression profiling guided by common retroviral insertion sites reveals novel and cell type specific cancer genes in leukemia. Blood. 2008;111:790–9.

    Article  CAS  PubMed  Google Scholar 

  8. Cooney E, Bi W, Schlesinger AE, Vinson S, Potocki L. Novel EED mutation in patient with Weaver syndrome. Am J Med Genet A. 2017;173:541–5.

    Article  CAS  PubMed  Google Scholar 

  9. Gibson WT, Hood RL, Zhan SH, Bulman DE, Fejes AP, Moore R, et al. Mutations in EZH2 cause Weaver syndrome. Am J Hum Genet. 2012;90:110–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Schumacher A, Faust C, Magnuson T. Positional cloning of a global regulator of anterior-posterior patterning in mice. Nature. 1996;384:648.

    Article  CAS  PubMed  Google Scholar 

  11. Hirabayashi Y, Suzki N, Tsuboi M, Endo TA, Toyoda T, Shinga J, et al. Polycomb limits the neurogenic competence of neural precursor cells to promote astrogenic fate transition. Neuron. 2009;63:600–13.

    Article  CAS  PubMed  Google Scholar 

  12. Wang J, Yang L, Dong C, Wang J, Xu L, Qiu Y, et al. EED-mediated histone methylation is critical for CNS myelination and remyelination by inhibiting WNT, BMP, and senescence pathways. Sci Adv. 2020;6:eaaz6477.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Li Q, Barres BA. Microglia and macrophages in brain homeostasis and disease. Nat Rev Immunol. 2018;18:225–42.

    Article  CAS  PubMed  Google Scholar 

  14. Madore C, Yin Z, Leibowitz J, Butovsky O. Microglia, lifestyle stress, and neurodegeneration. Immunity. 2020;52:222–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Nayak D, Roth TL, McGavern DB. Microglia development and function. Annu Rev Immunol. 2014;32:367–402.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Masuda T, Sankowski R, Staszewski O, Bottcher C, Amann L, Sagar, et al. Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution. Nature. 2019;566:388–92.

    Article  CAS  PubMed  Google Scholar 

  17. Doorn KJ, Breve JJ, Drukarch B, Boddeke HW, Huitinga I, Lucassen PJ, et al. Brain region-specific gene expression profiles in freshly isolated rat microglia. Front Cell Neurosci. 2015;9:84.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Li Q, Cheng Z, Zhou L, Darmanis S, Neff NF, Okamoto J, et al. Developmental heterogeneity of microglia and brain myeloid cells revealed by deep single-cell RNA sequencing. Neuron. 2019;101:207–23 e210.

    Article  CAS  PubMed  Google Scholar 

  19. Masuda T, Sankowski R, Staszewski O, Prinz M. Microglia heterogeneity in the single-cell era. Cell Rep. 2020;30:1271–81.

    Article  CAS  PubMed  Google Scholar 

  20. Grabert K, Michoel T, Karavolos MH, Clohisey S, Baillie JK, Stevens MP, et al. Microglial brain region-dependent diversity and selective regional sensitivities to aging. Nat Neurosci. 2016;19:504–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Hammond TR, Dufort C, Dissing-Olesen L, Giera S, Young A, Wysoker A, et al. Single-cell RNA sequencing of microglia throughout the mouse lifespan and in the injured brain reveals complex cell-state changes. Immunity. 2019;50:253–71 e256.

    Article  CAS  PubMed  Google Scholar 

  22. Tan YL, Yuan Y, Tian L. Microglial regional heterogeneity and its role in the brain. Mol Psychiatry. 2020;25:351–67.

    Article  PubMed  Google Scholar 

  23. De Biase LM, Bonci A. Region-specific phenotypes of microglia: the role of local regulatory cues. Neuroscientist. 2019;25:314–33.

    Article  PubMed  Google Scholar 

  24. Ayata P, Badimon A, Strasburger HJ, Duff MK, Montgomery SE, Loh YE, et al. Epigenetic regulation of brain region-specific microglia clearance activity. Nat Neurosci. 2018;21:1049–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Matcovitch-Natan O, Winter DR, Giladi A, Vargas Aguilar S, Spinrad A, Sarrazin S, et al. Microglia development follows a stepwise program to regulate brain homeostasis. Science. 2016;353:aad8670.

    Article  PubMed  CAS  Google Scholar 

  26. Zhan Y, Paolicelli RC, Sforazzini F, Weinhard L, Bolasco G, Pagani F, et al. Deficient neuron-microglia signaling results in impaired functional brain connectivity and social behavior. Nat Neurosci. 2014;17:400–6.

    Article  CAS  PubMed  Google Scholar 

  27. Paolicelli RC, Bolasco G, Pagani F, Maggi L, Scianni M, Panzanelli P, et al. Synaptic pruning by microglia is necessary for normal brain development. Science. 2011;333:1456–8.

    Article  CAS  PubMed  Google Scholar 

  28. Filipello F, Morini R, Corradini I, Zerbi V, Canzi A, Michalski B, et al. The microglial innate immune receptor TREM2 is required for synapse elimination and normal brain connectivity. Immunity. 2018;48:979–91 e978.

    Article  CAS  PubMed  Google Scholar 

  29. Keren-Shaul H, Spinrad A, Weiner A, Matcovitch-Natan O, Dvir-Szternfeld R, Ulland TK, et al. A unique microglia type associated with restricting development of Alzheimer’s disease. Cell. 2017;169:1276–90 e1217.

    Article  CAS  PubMed  Google Scholar 

  30. Krasemann S, Madore C, Cialic R, Baufeld C, Calcagno N, El Fatimy R, et al. The TREM2-APOE pathway drives the transcriptional phenotype of dysfunctional microglia in neurodegenerative diseases. Immunity. 2017;47:566–81 e569.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Oakley H, Cole SL, Logan S, Maus E, Shao P, Craft J, et al. Intraneuronal beta-amyloid aggregates, neurodegeneration, and neuron loss in transgenic mice with five familial Alzheimer’s disease mutations: potential factors in amyloid plaque formation. J Neurosci. 2006;26:10129–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Griciuc A, Patel S, Federico AN, Choi SH, Innes BJ, Oram MK, et al. TREM2 acts downstream of CD33 in modulating microglial pathology in Alzheimer’s disease. Neuron. 2019;103:820–35 e827.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lavarone E, Barbieri CM, Pasini D. Dissecting the role of H3K27 acetylation and methylation in PRC2 mediated control of cellular identity. Nat Commun. 2019;10:1679.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. Wu Y, Dissing-Olesen L, MacVicar BA, Stevens B. Microglia: dynamic mediators of synapse development and plasticity. Trends Immunol. 2015;36:605–13.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  35. Wake H, Moorhouse AJ, Jinno S, Kohsaka S, Nabekura J. Resting microglia directly monitor the functional state of synapses in vivo and determine the fate of ischemic terminals. J Neurosci. 2009;29:3974–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Nimmerjahn A, Kirchhoff F, Helmchen F. Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science. 2005;308:1314–8.

    Article  CAS  PubMed  Google Scholar 

  37. Wang C, Yue H, Hu Z, Shen Y, Ma J, Li J, et al. Microglia mediate forgetting via complement-dependent synaptic elimination. Science. 2020;367:688–94.

    Article  CAS  PubMed  Google Scholar 

  38. Andoh M, Koyama R. Microglia regulate synaptic development and plasticity. Dev Neurobiol. 2021;81:568–90.

    Article  PubMed  PubMed Central  Google Scholar 

  39. McCarthy MM. Location, location, location: microglia are where they live. Neuron. 2017;95:233–5.

    Article  CAS  PubMed  Google Scholar 

  40. Hickman SE, Kingery ND, Ohsumi TK, Borowsky ML, Wang LC, Means TK, et al. The microglial sensome revealed by direct RNA sequencing. Nat Neurosci. 2013;16:1896–905.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Gosselin D, Link VM, Romanoski CE, Fonseca GJ, Eichenfield DZ, Spann NJ, et al. Environment drives selection and function of enhancers controlling tissue-specific macrophage identities. Cell. 2014;159:1327–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Peng H, Geil Nickell CR, Chen KY, McClain JA, Nixon K. Increased expression of M1 and M2 phenotypic markers in isolated microglia after four-day binge alcohol exposure in male rats. Alcohol. 2017;62:29–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Kobayashi K, Imagama S, Ohgomori T, Hirano K, Uchimura K, Sakamoto K, et al. Minocycline selectively inhibits M1 polarization of microglia. Cell Death Dis. 2013;4:e525.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, et al. Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science. 2010;330:841–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Nikodemova M, Kimyon RS, De I, Small AL, Collier LS, Watters JJ. Microglial numbers attain adult levels after undergoing a rapid decrease in cell number in the third postnatal week. J Neuroimmunol. 2015;278:280–8.

    Article  CAS  PubMed  Google Scholar 

  46. Eich ML, Athar M, Ferguson JE 3rd, Varambally S. EZH2-targeted therapies in cancer: hype or a reality. Cancer Res. 2020;80:5449–58.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Laugesen A, Hojfeldt JW, Helin K. Molecular mechanisms directing PRC2 recruitment and H3K27 methylation. Mol Cell. 2019;74:8–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Zhang F, Yan Y, Cao X, Zhang J, Li Y, Guo C. Methylation of microRNA-338-5p by EED promotes METTL3-mediated translation of oncogene CDCP1 in gastric cancer. Aging. 2021;13:12224–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Klus P, Cirillo D, Botta Orfila T, Gaetano Tartaglia G. Neurodegeneration and cancer: where the disorder prevails. Sci Rep. 2015;5:15390.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Lanni C, Masi M, Racchi M, Govoni S. Cancer and Alzheimer’s disease inverse relationship: an age-associated diverging derailment of shared pathways. Mol Psychiatry. 2021;26:280–95.

    Article  CAS  PubMed  Google Scholar 

  51. Musicco M, Adorni F, Di Santo S, Prinelli F, Pettenati C, Caltagirone C, et al. Inverse occurrence of cancer and Alzheimer disease: a population-based incidence study. Neurology. 2013;81:322–8.

    Article  PubMed  Google Scholar 

  52. Ibanez K, Boullosa C, Tabares-Seisdedos R, Baudot A, Valencia A. Molecular evidence for the inverse comorbidity between central nervous system disorders and cancers detected by transcriptomic meta-analyses. PLoS Genet. 2014;10:e1004173.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  53. Cheray M, Stratoulias V, Joseph B, Grabert K. The rules of engagement: do microglia seal the fate in the inverse relation of Glioma and Alzheimer’s disease? Front Cell Neurosci. 2019;13:522.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Liu PP, Tang GB, Xu YJ, Zeng YQ, Zhang SF, Du HZ, et al. MiR-203 interplays with polycomb repressive complexes to regulate the proliferation of neural stem/progenitor cells. Stem Cell Rep. 2017;9:190–202.

    Article  CAS  Google Scholar 

  55. Dai SK, Liu PP, Du HZ, Liu X, Xu YJ, Liu C, et al. Histone crotonylation regulates neural stem cell fate decisions by activating bivalent promoters. EMBO Rep. 2021; 22:e52023.

  56. Tang GB, Zeng YQ, Liu PP, Mi TW, Zhang SF, Dai SK, et al. The histone H3K27 demethylase UTX regulates synaptic plasticity and cognitive behaviors in mice. Front Mol Neurosci. 2017;10:267.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  57. Cheng Y, Wang ZM, Tan W, Wang X, Li Y, Bai B, et al. Partial loss of psychiatric risk gene Mir137 in mice causes repetitive behavior and impairs sociability and learning via increased Pde10a. Nat Neurosci. 2018;21:1689–703.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Saura J, Tusell JM, Serratosa J. High-yield isolation of murine microglia by mild trypsinization. Glia. 2003;44:183–9.

    Article  PubMed  Google Scholar 

  59. Gao X, Enikolopov G, Chen J. Moderate traumatic brain injury promotes proliferation of quiescent neural progenitors in the adult hippocampus. Exp Neurol. 2009;219:516–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We appreciate the editors and five anonymous reviewers for the in-depth comments, suggestions, and corrections, which have greatly improved the paper. This work was supported by the National Key Research and Development Program of China Project (2018YFA0108001/2021YFA1101402), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16010300), the National Science Foundation of China (32170808), and the Open Project Program of State Key Laboratory of Stem Cell and Reproductive Biology.

Author information

Authors and Affiliations

  1. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China

    Ying-Ying Wang, Yu-Sen Deng, Shang-Kun Dai, Ting-Wei Mi, Pei-Pei Liu, Cong Liu, Bao-Dong He, Xuan-Cheng He, Hong-Zhen Du, Chang-Mei Liu & Zhao-Qian Teng

  2. Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China

    Ying-Ying Wang, Yu-Sen Deng, Shang-Kun Dai, Bao-Dong He, Chang-Mei Liu & Zhao-Qian Teng

  3. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China

    Ying-Ying Wang, Yu-Sen Deng, Shang-Kun Dai, Pei-Pei Liu, Cong Liu, Bao-Dong He, Xuan-Cheng He, Hong-Zhen Du, Chang-Mei Liu & Zhao-Qian Teng

  4. Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, 100101, China

    Ying-Ying Wang, Pei-Pei Liu, Cong Liu, Xuan-Cheng He, Hong-Zhen Du, Chang-Mei Liu & Zhao-Qian Teng

  5. Innovation Center for Neurological Disorders, Department of Neurology, Xuanwu Hospital, Capital Medical University, Beijing, 100053, China

    Rui-Yang Li, Han-Chen Yang & Yi Tang

Authors
  1. Ying-Ying Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yu-Sen Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shang-Kun Dai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ting-Wei Mi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rui-Yang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pei-Pei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Cong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Bao-Dong He
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xuan-Cheng He
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hong-Zhen Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Han-Chen Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yi Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Chang-Mei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Zhao-Qian Teng
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ZQT, CML, and YYW, conceived; ZQT, CML, and YT, supervised this study; YYW, ZQT, and CML, designed experiments; YYW, YSD, SKD, TWM, PPL, CL, RYL, BDH, XCH, HZD, and HCY, performed and analyzed experiments; SKD and YYW, performed bioinformatics analysis. YYW and ZQT, prepared figures and wrote the paper.

Corresponding authors

Correspondence to Yi Tang, Chang-Mei Liu or Zhao-Qian Teng.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, YY., Deng, YS., Dai, SK. et al. Loss of microglial EED impairs synapse density, learning, and memory. Mol Psychiatry 27, 2999–3009 (2022). https://doi.org/10.1038/s41380-022-01576-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41380-022-01576-w

Search

Advanced search

Quick links