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Retinal Degeneration pp 153–164Cite as

Fate Mapping In Vivo to Distinguish Bona Fide Microglia Versus Recruited Monocyte-Derived Macrophages in Retinal Disease

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1834))

Abstract

With the new understanding that adult microglia in mice have embryonic origins and are maintained in situ throughout life, it has become pertinent to now understand how these unique cells differ from monocyte-derived macrophages. The latter are recruited into the neural retina (and elsewhere in CNS) in certain diseased states, such as in various forms of retinal degeneration. However, phenotypic markers expressed by microglia and monocyte-derived macrophages largely overlap, thereby making it technically challenging to distinguish the two cell types in disease. To address this problem in mice, we have established an in vivo fate mapping system that enables distinguishing these two cell types in retinal disease models. Our approach leverages the seminal work that originally developed Cx3cr1-CreER mice and is based on commercially available mouse strains. Here, we detail our protocol and how to apply this fate mapping method paired with flow cytometry (or immunohistochemistry) to faithfully distinguish and examine microglia vs. monocyte-derived macrophages in a mutually exclusive manner. This approach will henceforth empower new efforts to identify functional specializations of these two populations in the pathobiology of retinal degenerative diseases and possibly other conditions of the retina where monocyte recruitment is observed, such as in glaucoma, diabetic retinopathy, ischemia reperfusion, retinal detachment, and so on.

Grant Information: R01EY021798 (Saban), P30EY005722 (Saban), Bright Focus MDR (Saban), and F32EY025557 (Reyes)

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Author information

Authors and Affiliations

  1. Department of Ophthalmology, Duke University School of Medicine, Durham, NC, USA

    Nancy J. Reyes, Rose Mathew & Daniel R. Saban

Authors
  1. Nancy J. Reyes
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  2. Rose Mathew
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  3. Daniel R. Saban
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Corresponding author

Correspondence to Daniel R. Saban .

Editor information

Editors and Affiliations

  1. Institute of Human Genetics, University of Regensburg, Regensburg, Germany

    Bernhard H. F. Weber

  2. Department of Ophthalmology, University of Cologne, Cologne, Germany

    Thomas Langmann

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Reyes, N.J., Mathew, R., Saban, D.R. (2019). Fate Mapping In Vivo to Distinguish Bona Fide Microglia Versus Recruited Monocyte-Derived Macrophages in Retinal Disease. In: Weber, B.H.F., Langmann, T. (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_11

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  • DOI: https://doi.org/10.1007/978-1-4939-8669-9_11

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8668-2

  • Online ISBN: 978-1-4939-8669-9

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