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Graefe's Archive for Clinical and Experimental Ophthalmology

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01-02-2018 | Basic Science

Spatial distribution of CD115⁺ and CD11b⁺ cells and their temporal activation during oxygen-induced retinopathy in mice

Authors: Claudia Brockmann, Sabrina Dege, Sergio Crespo-Garcia, Norbert Kociok, Tobias Brockmann, Olaf Strauß, Antonia M. Joussen

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 2/2018

Abstract

Purpose

The model of oxygen-induced retinopathy (OIR) is widely used to analyze pathomechanisms in retinal neovascularization. Previous studies have shown that macrophages (MP) play a key role in vessel formation in OIR, the influence of microglia (MG) having been discussed. The aim of our study was to analyze the spatial and temporal distribution and activation of MP/MG expressing CD115 and CD11b during the process of neovascularization in OIR.

Methods

We used MacGreen mice expressing the green fluorescence protein (GFP) under the promoter for CD115. CD115⁺ cells were investigated in vivo by scanning laser ophthalmoscopy at postnatal days (P) 17 and 21 in MacGreen mice with OIR (75% oxygen from P7 to P12), and were compared to MacGreen room-air controls. In addition MP/MG were examined ex vivo using immunohistochemistry for CD11b⁺ detection on retinal flatmounts at P14, P17, and P21 of wild type mice with OIR.

Results

In-vivo imaging revealed the highest density of activated MP/MG in tuft areas at P17 of MacGreen mice with OIR. Tufts and regions with a high density of CD115⁺ cells were detected close to veins, rather to arteries. In peripheral, fully vascularized areas, the distribution of CD115⁺ cells in MacGreen mice with OIR was similar to MacGreen room-air controls. Correspondingly, immunohistochemical analyses of retinal flatmounts from wild type mice with OIR induction revealed that the number of CD11b⁺ cells significantly varies between vascular, avascular, and tuft areas as well as between the retinal layers. Activated CD11b⁺ cells were almost exclusively found in avascular areas and tufts of wild type mice with OIR induction; here, the proportion of activated cells related to the total number of CD11b⁺ cells remained stable over the course of time.

Conclusions

Using two different approaches to monitor MP/MG cells, our findings demonstrated that MP/MG concentrate within pathologically vascularized areas during OIR. We were able to clarify that reactive changes of CD11b⁺ cell distribution to OIR primarily occur in the deep retinal layers. Furthermore, we found the highest proportion of activated CD11b⁺ cells in regions with pathologic neovascularization processes. Our findings support previous reports about activated MP/MG guiding revascularization in avascular areas and playing a key role in the formation and regression of neovascular tufts.

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Title: Spatial distribution of CD115+ and CD11b+ cells and their temporal activation during oxygen-induced retinopathy in mice
Authors: Claudia Brockmann
Sabrina Dege
Sergio Crespo-Garcia
Norbert Kociok
Tobias Brockmann
Olaf Strauß
Antonia M. Joussen
Publication date: 01-02-2018
Publisher: Springer Berlin Heidelberg
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 2/2018
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-017-3845-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Spatial distribution of CD115⁺ and CD11b⁺ cells and their temporal activation during oxygen-induced retinopathy in mice

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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