Universität zu Köln

Lueckoff, Anika, Scholz, Rebecca, Sennlaub, Florian ORCID: 0000-0003-4412-1341, Xu, Heping ORCID: 0000-0003-4000-931X and Langmann, Thomas (2017). Comprehensive analysis of mouse retinal mononuclear phagocytes. Nat. Protoc., 12 (6). S. 1136 - 1151. LONDON: NATURE PUBLISHING GROUP. ISSN 1750-2799

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Abstract

The innate immune system is activated in a number of degenerative and inflammatory retinal disorders such as age-related macular degeneration (AMD). Retinal microglia, choroidal macrophages, and recruited monocytes, collectively termed 'retinal mononuclear phagocytes', are critical determinants of ocular disease outcome. Many publications have described the presence of these cells in mouse models for retinal disease; however, only limited aspects of their behavior have been uncovered, and these have only been uncovered using a single detection method. The workflow presented here describes a comprehensive analysis strategy that allows characterization of retinal mononuclear phagocytes in vivo and in situ. We present standardized working steps for scanning laser ophthalmoscopy of microglia from MacGreen reporter mice (mice expressing the macrophage colony-stimulating factor receptor GFP transgene throughout the mononuclear phagocyte system), quantitative analysis of Iba1-stained retinal sections and flat mounts, CD11b-based retinal flow cytometry, and qRTRT-PCRPCRPCR analysis of key microglia markers. The protocol can be completed within 3 d, and we present data from retinas treated with laser-induced choroidal neovascularization (CNCNV), bright white-light exposure, and Fam161a-associated inherited retinal degeneration. The assays can be applied to any of the existing mouse models for retinal disorders and may be valuable for documenting immune responses in studies for immunomodulatory therapies.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Lueckoff, Anika UNSPECIFIED UNSPECIFIED UNSPECIFIED Scholz, Rebecca UNSPECIFIED UNSPECIFIED UNSPECIFIED Sennlaub, Florian UNSPECIFIED orcid.org/0000-0003-4412-1341 UNSPECIFIED Xu, Heping UNSPECIFIED orcid.org/0000-0003-4000-931X UNSPECIFIED Langmann, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-230459
DOI:	10.1038/nprot.2017.032
Journal or Publication Title:	Nat. Protoc.
Volume:	12
Number:	6
Page Range:	S. 1136 - 1151
Date:	2017
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	1750-2799
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language INDUCED CHOROIDAL NEOVASCULARIZATION; ACTIVATED MICROGLIA/MACROPHAGE; PHOTORECEPTOR DEGENERATION; MACULAR DEGENERATION; IN-VIVO; MICROGLIAL PHAGOCYTOSIS; SUBRETINAL INFLAMMATION; RETINITIS-PIGMENTOSA; AGE; MODEL Multiple languages Biochemical Research Methods Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/23045