Tmem119-EGFP and Tmem119-CreERT2 Transgenic Mice for Labeling and Manipulating Microglia

Abstract

Microglia are specialized brain-resident macrophages with important functions in health and disease. To improve our understanding of these cells, the research community needs genetic tools to identify and control them in a manner that distinguishes them from closely related cell types. We have targeted the recently discovered microglia-specific Tmem119 gene to generate knock-in mice expressing EGFP (JAX#031823) or CreERT2 (JAX#031820) for the identification and manipulation of microglia, respectively. Genetic characterization of the locus and qPCR-based analysis demonstrate correct positioning of the transgenes and intact expression of endogenous Tmem119 in the knock-in mouse models. Immunofluorescence analysis further shows that parenchymal microglia, but not other brain macrophages, are completely and faithfully labeled in the EGFP-line at different time points of development. Flow cytometry indicates highly selective expression of EGFP in CD11b + CD45lo microglia. Similarly, immunofluorescence and flow cytometry analyses using a Cre-dependent reporter mouse line demonstrate activity of CreERT2 primarily in microglia upon tamoxifen administration with the caveat of activity in leptomeningeal cells. Finally, flow cytometric analyses reveal absence of EGFP expression and minimal activity of CreERT2 in blood monocytes of the Tmem119-EGFP and Tmem119-CreERT2 lines, respectively. These new transgenic lines extend the microglia toolbox by providing the currently most specific genetic labeling and control over these cells in the myeloid compartment of mice.

Shown for reading. Its publisher allows quoting it, not republishing it.

References 39

1. 1.

   Aguzzi A, Zhu C (2017 ) Microglia in prion diseases. J Clin Invest 127:3230–3239.

   Not in our library10.1172/jci90605

2. 2.

   Bennett ML, Bennett FC, Liddelow SA, Ajami B, Zamanian JL, Fernhoff NB, Mulinyawe SB, Bohlen CJ, Adil A, Tucker A, Weissman IL, Chang EF, Li G, Grant GA, Hayden Gephart MG, Barres BA (2016 ) New tools for studying microglia in the mouse and human CNS. Proc Natl Acad Sci U S A 113:E1738–E1746.

   Not in our library10.1073/pnas.1525528113

3. 3.

   Butovsky O, Jedrychowski MP, Moore CS, Cialic R, Lanser AJ, Gabriely G, Koeglsperger T, Dake B, Wu PM, Doykan CE, Fanek Z, Liu L, Chen Z, Rothstein JD, Ransohoff RM, Gygi SP, Antel JP, Weiner HL (2014 ) Identification of a unique TGF-β–dependent molecular and functional signature in microglia. Nat Neurosci 17:131.

   Not in our library10.1038/nn.3599

4. 4.

   Buttgereit A, Lelios I, Yu X, Vrohlings M, Krakoski NR, Gautier EL, Nishinakamura R, Becher B, Greter M (2016 ) Sall1 is a transcriptional regulator defining microglia identity and function. Nat Immunol 17:1397–1406.

   Not in our library10.1038/ni.3585

5. 5.

   Clausen B, Burkhardt C, Reith W, Renkawitz R, Förster I (1999 ) Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res 8:265–277.

   Not in our library10.1023/a:1008942828960

6. 6.

   Davalos D, Grutzendler J, Yang G, Kim JV, Zuo Y, Jung S, Littman DR, Dustin ML, Gan WB (2005 ) ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci 8:752.

   Not in our library10.1038/nn1472

7. 7.

   Decimo I, Fumagalli G, Berton V, Krampera M, Bifari F (2012 ) Meninges: from protective membrane to stem cell niche. Am J Stem Cells 1:92–105.

   Not in our library

8. 8.

   Ferron M, Vacher J (2005 ) Targeted expression of Cre recombinase in macrophages and osteoclasts in transgenic mice. Genesis 41:138–145.

   Not in our library10.1002/gene.20108

9. 9.

   Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, Mehler MF, Conway SJ, Ng LG, Stanley ER (2010 ) Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 330:841–845.

   Not in our library10.1126/science.1194637

10. 10.

    Goldmann T, Wieghofer P, Müller PF, Wolf Y, Varol D, Yona S, Brendecke SM, Kierdorf K, Staszewski O, Datta M, Luedde T, Heikenwalder M, Jung S, Prinz M (2013 ) A new type of microglia gene targeting shows TAK1 to be pivotal in CNS autoimmune inflammation. Nat Neurosci 16:1618.

    Not in our library10.1038/nn.3531

11. 11.

    Goldmann T, Wieghofer P, Jordão MJ, Prutek F, Hagemeyer N, Frenzel K, Amann L, Staszewski O, Kierdorf K, Krueger M, Locatelli G, Hochgerner H, Zeiser R, Epelman S, Geissmann F, Priller J, Rossi FM, Bechmann I, Kerschensteiner M, Linnarsson S, et al. (2016 ) Origin, fate and dynamics of macrophages at central nervous system interfaces. Nat Immunol 17:797.

    Not in our library10.1038/ni.3423

12. 12.

    Haimon Z, Volaski A, Orthgiess J, Boura-Halfon S, Varol D, Shemer A, Yona S, Zuckerman B, David E, Chappell-Maor L, Bechmann I, Gericke M, Ulitsky I, Jung S (2018 ) Re-evaluating microglia expression profiles using RiboTag and cell isolation strategies. Nat Immunol 19:636–644.

    Not in our library10.1038/s41590-018-0110-6

13. 13.

    Hirasawa T, Ohsawa K, Imai Y, Ondo Y, Akazawa C, Uchino S, Kohsaka S (2005 ) Visualization of microglia in living tissues using Iba1-EGFP transgenic mice. J Neurosci Res 81:357–362.

    Not in our library10.1002/jnr.20480

14. 14.

    Itagaki S, McGeer P, Akiyama H, Zhu S, Selkoe D (1989 ) Relationship of microglia and astrocytes to amyloid deposits of Alzheimer disease. J Neuroimmunol 24:173–182.

    Not in our library10.1016/0165-5728(89)90115-x

15. 15.

    Keren-Shaul H, Spinrad A, Weiner A, Matcovitch-Natan O, Dvir-Szternfeld R, Ulland TK, David E, Baruch K, Lara-Astaiso D, Toth B, Itzkovitz S, Colonna M, Schwartz M, Amit I (2017 ) A unique microglia type associated with restricting development of Alzheimer's disease. Cell 169:1276–1290.e17.

    Not in our library10.1016/j.cell.2017.05.018

16. 16.

    Kim WK, Alvarez X, Fisher J, Bronfin B, Westmoreland S, McLaurin J, Williams K (2006 ) CD163 identifies perivascular macrophages in normal and viral encephalitic brains and potential precursors to perivascular macrophages in blood. Am J Pathol 168:822–834.

    Not in our library10.2353/ajpath.2006.050215

17. 17.

    Lawson LJ, Perry VH, Dri P, Gordon S (1990 ) Heterogeneity in the distribution and morphology of microglia in the normal adult mouse brain. Neuroscience 39:151–170.

    Not in our library10.1016/0306-4522(90)90229-w

18. 18.

    Lee S, Varvel NH, Konerth ME, Xu G, Cardona AE, Ransohoff RM, Lamb BT (2010 ) CX3CR1 deficiency alters microglial activation and reduces beta-amyloid deposition in two Alzheimer's disease mouse models. Am J Pathol 177:2549–2562.

    Not in our library10.2353/ajpath.2010.100265

19. 19.

    Li Q, Cheng Z, Zhou L, Darmanis S, Neff NF, Okamoto J, Gulati G, Bennett ML, Sun LO, Clarke LE, Marschallinger J, Yu G, Quake SR, Wyss-Coray T, Barres BA (2019 ) Developmental heterogeneity of microglia and brain myeloid cells revealed by deep single-cell RNA sequencing. Neuron 101:207–223.e10.

    Not in our library10.1016/j.neuron.2018.12.006

20. 20.

    Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H (2010 ) A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13:133–140.

    Not in our library10.1038/nn.2467

21. 21.

    Mathys H, Adaikkan C, Gao F, Young JZ, Manet E, Hemberg M, De Jager PL, Ransohoff RM, Regev A, Tsai LH (2017 ) Temporal tracking of microglia activation in neurodegeneration at single-cell resolution. Cell Rep 21:366–380.

    Not in our library10.1016/j.celrep.2017.09.039

22. 22.

    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 65:375–387.

    Not in our library10.1002/glia.23097

23. 23.

    O’Loughlin E, Madore C, Lassmann H, Butovsky O (2018 ) Microglial phenotypes and functions in multiple sclerosis. Cold Spring Harb Perspect Med 8:a028993

    Not in our library10.1101/cshperspect.a028993

24. 24.

    Paolicelli RC, Bolasco G, Pagani F, Maggi L, Scianni M, Panzanelli P, Giustetto M, Ferreira TA, Guiducci E, Dumas L, Ragozzino D, Gross CT (2011 ) Synaptic pruning by microglia is necessary for normal brain development. Science 333:1456–1458.

    Not in our library10.1126/science.1202529

25. 25.

    Parkhurst CN, Yang G, Ninan I, Savas JN, Yates JR, Lafaille JJ, Hempstead BL, Littman DR, Gan WB (2013 ) Microglia promote learning-dependent synapse formation through brain-derived neurotrophic factor. Cell 155:1596–1609.

    Not in our library10.1016/j.cell.2013.11.030

26. 26.

    Prinz M, Priller J (2014 ) Microglia and brain macrophages in the molecular age: from origin to neuropsychiatric disease. Nat Rev Neurosci 15:300.

    Not in our library10.1038/nrn3722

27. 27.

    Ransohoff RM (2016 ) How neuroinflammation contributes to neurodegeneration. Science 353:777–783.

    Not in our library10.1126/science.aag2590

28. 28.

    Rogers JT, Morganti JM, Bachstetter AD, Hudson CE, Peters MM, Grimmig BA, Weeber EJ, Bickford PC, Gemma C (2011 ) CX3CR1 deficiency leads to impairment of hippocampal cognitive function and synaptic plasticity. J Neurosci 31:16241–16250.

    Not in our library10.1523/jneurosci.3667-11.2011

29. 29.

    Samokhvalov IM, Samokhvalova NI, Nishikawa SI (2007 ) Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446:1056.

    Not in our library10.1038/nature05725

30. 30.

    Sasmono RT, Oceandy D, Pollard JW, Tong W, Pavli P, Wainwright BJ, Ostrowski MC, Himes SR, Hume DA (2003 ) A macrophage colony-stimulating factor receptor–green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse. Blood 101:1155–1163.

    Not in our library10.1182/blood-2002-02-0569

31. 31.

    Satoh JI, Kino Y, Asahina N, Takitani M, Miyoshi J, Ishida T, Saito Y (2016 ) TMEM119 marks a subset of microglia in the human brain. Neuropathology 36:39–49.

    Not in our library10.1111/neup.12235

32. 32.

    Schafer DP, Lehrman EK, Kautzman AG, Koyama R, Mardinly AR, Yamasaki R, Ransohoff RM, Greenberg ME, Barres BA, Stevens B (2012 ) Microglia sculpt postnatal neural circuits in an activity and complement-dependent manner. Neuron 74:691–705.

    Not in our library10.1016/j.neuron.2012.03.026

33. 33.

    Sierra A, Encinas JM, Deudero JJ, Chancey JH, Enikolopov G, Overstreet-Wadiche LS, Tsirka SE, Maletic-Savatic M (2010 ) Microglia shape adult hippocampal neurogenesis through apoptosis-coupled phagocytosis. Cell Stem Cell 7:483–495.

    Not in our library10.1016/j.stem.2010.08.014

34. 34.

    Stevens B, Allen NJ, Vazquez LE, Howell GR, Christopherson KS, Nouri N, Micheva KD, Mehalow AK, Huberman AD, Stafford B, Sher A, Litke Alan M, Lambris JD, Smith SJ, John SWM, Barres BA (2007 ) The classical complement cascade mediates CNS synapse elimination. Cell 131:1164–1178.

    Not in our library10.1016/j.cell.2007.10.036

35. 35.

    Ueno M, Fujita Y, Tanaka T, Nakamura Y, Kikuta J, Ishii M, Yamashita T (2013 ) Layer V cortical neurons require microglial support for survival during postnatal development. Nat Neurosci 16:543–551.

    Not in our library10.1038/nn.3358

36. 36.

    Vanlandewijck M, He L, Mäe MA, Andrae J, Ando K, Del Gaudio F, Nahar K, Lebouvier T, Laviña B, Gouveia L, Sun Y, Raschperger E, Räsänen M, Zarb Y, Mochizuki N, Keller A, Lendahl U, Betsholtz C (2018 ) A molecular atlas of cell types and zonation in the brain vasculature. Nature 554:475.

    Not in our library10.1038/nature25739

37. 37.
    Microglia remodel synapses by presynaptic trogocytosis and spine head filopodia induction · 2018

    No notice
38. 38.

    Wieghofer P, Prinz M (2016 ) Genetic manipulation of microglia during brain development and disease. Biochimica et Biophysica Acta 1862:299–309.

    Not in our library10.1016/j.bbadis.2015.09.019

39. 39.

    Yona S, Kim KW, Wolf Y, Mildner A, Varol D, Breker M, Strauss-Ayali D, Viukov S, Guilliams M, Misharin A, Hume David A, Perlman H, Malissen B, Zelzer E, Jung S (2013 ) Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis. Immunity 38:79–91.

    Not in our library10.1016/j.immuni.2012.12.001

Cited by papers we hold 9

• Glioma-associated microglia/macrophages (GAMs) in glioblastoma: Immune function in the tumor microenvironment and implications for immunotherapy · 2023
  Not checked
• Transcriptional and epigenetic decoding of the microglial aging process · 2023
  Corrected
• Neuroinflammation, Microglia and Implications for Retinal Ganglion Cell Survival and Axon Regeneration in Traumatic Optic Neuropathy · 2022
  No notice
• Microglial/Macrophage polarization and function in brain injury and repair after stroke · 2021
  No notice
• ROS Generation in Microglia: Understanding Oxidative Stress and Inflammation in Neurodegenerative Disease · 2020
  No notice
• A new genetic strategy for targeting microglia in development and disease · 2020
  No notice
• Regulation of microglial TMEM119 and P2RY12 immunoreactivity in multiple sclerosis white and grey matter lesions is dependent on their inflammatory environment · 2019
  No notice
• Beneficial and Detrimental Remodeling of Glial Connexin and Pannexin Functions in Rodent Models of Nervous System Diseases · 2019
  Not checked
• Moving beyond the glial scar for spinal cord repair · 2019
  No notice

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.