Microglia pp 325-336 | Cite as

Transcriptomics and Live Imaging to Define Functional Phenotypes of Microglia in Pathological Human Tissue

  • Giampaolo Milior
  • Farah Chali
  • Tiffany Dos Santos
  • Juliette Royer
  • Richard Miles
  • Mélanie Morin-BrureauEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2034)


Human microglia, as those of rodents, may possess multiple functional phenotypes. Here we present protocols to determine elements of these microglial phenotypes obtained after therapeutic excision of brain tissue from patients with epilepsies of the temporal lobe and cortical gliomas. This technique permits to identify microglia, to determine their shape and expression of state-specific markers, to measure resting and induced motilities, to define a human microglial transcriptome, and to determine how it changes after a seizure.

Key words

Human microglia Transcriptomics Microglial markers Motility Epilepsy 


  1. 1.
    Boche D, Perry VH, Nicoll JAR (2013) Activation patterns of microglia and their identification in the human brain. Neuropathol Appl Neurobiol 39:3–18CrossRefGoogle Scholar
  2. 2.
    Galatro TF, Holtman IR, Lerario AM, Vainchtein ID, Brouwer N, Sola P, Veras MM, Pereira TF, Leite REP, Möller T, Wes PD, Sogayar MC, Laman JD, den Dunnen W, Pasqualucci CA, Oba-Shinjo SM, Boddeke EW, Marie SKN, Eggen BJL (2017) Transcriptomic analysis of purified human cortical microglia reveals age-associated changes. Nat Neurosci 20:1162–1171CrossRefGoogle Scholar
  3. 3.
    Bohlen CJ, Bennett FC, Tucker AF, Collins HY, Mulinyawe SB, Barres BA (2017) Diverse requirements for microglial survival, specification, and function revealed by defined-medium cultures. Neuron 94:759–773CrossRefGoogle Scholar
  4. 4.
    Réu P, Khosravi A, Bernard S, Mold JE, Salehpour M, Alkass K, Perl S, Tisdale J, Possnert G, Druid H, Frisén J (2017) The lifespan and turnover of microglia in the human brain. Cell Rep 20:779–784CrossRefGoogle Scholar
  5. 5.
    Kuhn A, Thu D, Waldvogel HJ, Faull RL, Luthi-Carter R (2011) Population-specific expression analysis (PSEA) reveals molecular changes in diseased brain. Nat Methods 8:945–947CrossRefGoogle Scholar
  6. 6.
    Morin-Brureau M, Milior G, Royer J, Chali F, LeDuigou C, Savary E, Blugeon C, Jourdren L, Akbar D, Dupont S, Navarro V, Baulac M, Bielle F, Mathon B, Clemenceau S, Miles R (2018) Microglial phenotypes in the human epileptic temporal lobe. Brain 141:3343–3360CrossRefGoogle Scholar
  7. 7.
    Mildner A, Huang H, Radke J, Stenzel W, Priller J (2017) P2Y12 receptor is expressed on human microglia under physiological conditions throughout development and is sensitive to neuroinflammatory diseases. Glia 65:375–387CrossRefGoogle Scholar
  8. 8.
    Bordey A, Spencer DD (2003) Chemokine modulation of high-conductance Ca2+-sensitive K+ currents in microglia from human hippocampi. Eur J Neurosci 18:2893–2898CrossRefGoogle Scholar
  9. 9.
    Schwendele B, Brawek B, Hermes M, Garaschuk O (2012) High-resolution in vivo imaging of microglia using a versatile nongenetically encoded marker. Eur J Immunol 42:2193–2196CrossRefGoogle Scholar
  10. 10.
    Dissing-Olesen L, MacVicar BA (2015) Fixation and immunolabeling of brain slices: SNAPSHOT method. Curr Protoc Neurosci 71(1.23):1–12PubMedGoogle Scholar
  11. 11.
    Jourdren L, Bernard M, Dillies MA, Le Crom S (2012) Eoulsan: a cloud computing-based framework facilitating high throughput sequencing analyses. Bioinformatics 28:1542–1543CrossRefGoogle Scholar
  12. 12.
    Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15:550CrossRefGoogle Scholar
  13. 13.
    Lacar B, Linker SB, Jaeger BN, Krishnaswami S, Barron J, Kelder M, Parylak SL, Paquola ACM, Venepally P, Novotny M, O'Connor C, Fitzpatrick C, Erwin JA, Hsu JY, Husband D, McConnell MJ, Lasken R, Gage FH (2016) Nuclear RNA-seq of single neurons reveals molecular signatures of activation. Nat Commun 7:11022CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Giampaolo Milior
    • 2
  • Farah Chali
    • 1
  • Tiffany Dos Santos
    • 1
  • Juliette Royer
    • 1
  • Richard Miles
    • 1
  • Mélanie Morin-Brureau
    • 1
    • 3
    Email author
  1. 1.Sorbonne Université, Inserm UMRS 938, Centre de Recherche Saint-Antoine, CRSAParisFrance
  2. 2.Neuroglial Interactions in Cerebral Physiopathology, Center for Interdisciplinary Research in Biology, Collège de FranceCNR UMR 7241, INSERM U1050, Labex Memolife, PSL Research UniversityParisFrance
  3. 3.Team Immune System and NeuroinflammationSorbonne Université, Inserm UMRS 938, Centre de Recherche Saint-Antoine, CRSAParisFrance

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