Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies
Abstract Reliable tools for macrophage identification in mouse tissues are critical for studies investigating inflammatory and reparative responses. Transgenic reporter mice and anti-macrophage antibodies have been used as “specific pan-macrophage” markers in many studies; however, organ-specific pa...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-08278-x |
| _version_ | 1818320970832674816 |
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| author | Bijun Chen Ruoshui Li Akihiko Kubota Linda Alex Nikolaos G. Frangogiannis |
| author_facet | Bijun Chen Ruoshui Li Akihiko Kubota Linda Alex Nikolaos G. Frangogiannis |
| author_sort | Bijun Chen |
| collection | DOAJ |
| description | Abstract Reliable tools for macrophage identification in mouse tissues are critical for studies investigating inflammatory and reparative responses. Transgenic reporter mice and anti-macrophage antibodies have been used as “specific pan-macrophage” markers in many studies; however, organ-specific patterns of expression and non-specific labeling of other cell types, such as fibroblasts, may limit their usefulness. Our study provides a systematic comparison of macrophage labeling patterns in normal and injured mouse tissues, using the CX3CR1 and CSF1R macrophage reporter lines and anti-macrophage antibodies. Moreover, we tested the specificity of macrophage antibodies using the fibroblast-specific PDGFR $$\mathrm{\alpha }$$ α reporter line. Mouse macrophages exhibit organ-specific differences in expression of macrophage markers. Hepatic macrophages are labeled for CSF1R, Mac2 and F4/80, but lack CX3CR1 expression, whereas in the lung, the CSF1R+/Mac2+/Mac3+ macrophage population is not labeled with F4/80. In the splenic red pulp, subpopulations of CSF1R+/F4/80+/Mac3+cells were labeled with Mac2, CX3CR1 and lysozyme M. In the kidney, Mac2, Mac3 and lysozyme M labeled a fraction of the CSF1R+ and CX3CR1+ macrophages, but also stained tubular epithelial cells. In normal hearts, the majority of CSF1R+ and CX3CR1+ cells were not detected with anti-macrophage antibodies. Myocardial infarction was associated with marked expansion of the CSF1R+ and CX3CR1+ populations that peaked during the proliferative phase of cardiac repair, and also expressed Mac2, Mac3 and lysozyme M. In normal mouse tissues, a small fraction of cells labeled with anti-macrophage antibodies were identified as PDGFR $$\mathrm{\alpha }$$ α + fibroblasts, using a reporter system. The population of PDGFR $$\mathrm{\alpha }$$ α + cells expressing macrophage markers expanded following injury, likely reflecting emergence of cellular phenotypes with both fibroblast and macrophage characteristics. In conclusion, mouse macrophages exhibit remarkable heterogeneity. Selection of the most appropriate markers for identification of macrophages in mouse tissues is dependent on the organ and the pathologic condition studied. |
| first_indexed | 2024-12-13T10:33:28Z |
| format | Article |
| id | doaj.art-3b7ea66225014cfbbd3e63a02dbf7432 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-13T10:33:28Z |
| publishDate | 2022-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-3b7ea66225014cfbbd3e63a02dbf74322022-12-21T23:50:47ZengNature PortfolioScientific Reports2045-23222022-03-0112111710.1038/s41598-022-08278-xIdentification of macrophages in normal and injured mouse tissues using reporter lines and antibodiesBijun Chen0Ruoshui Li1Akihiko Kubota2Linda Alex3Nikolaos G. Frangogiannis4The Wilf Family Cardiovascular Research Institute, Department of Medicine (Cardiology), Albert Einstein College of MedicineThe Wilf Family Cardiovascular Research Institute, Department of Medicine (Cardiology), Albert Einstein College of MedicineThe Wilf Family Cardiovascular Research Institute, Department of Medicine (Cardiology), Albert Einstein College of MedicineThe Wilf Family Cardiovascular Research Institute, Department of Medicine (Cardiology), Albert Einstein College of MedicineThe Wilf Family Cardiovascular Research Institute, Department of Medicine (Cardiology), Albert Einstein College of MedicineAbstract Reliable tools for macrophage identification in mouse tissues are critical for studies investigating inflammatory and reparative responses. Transgenic reporter mice and anti-macrophage antibodies have been used as “specific pan-macrophage” markers in many studies; however, organ-specific patterns of expression and non-specific labeling of other cell types, such as fibroblasts, may limit their usefulness. Our study provides a systematic comparison of macrophage labeling patterns in normal and injured mouse tissues, using the CX3CR1 and CSF1R macrophage reporter lines and anti-macrophage antibodies. Moreover, we tested the specificity of macrophage antibodies using the fibroblast-specific PDGFR $$\mathrm{\alpha }$$ α reporter line. Mouse macrophages exhibit organ-specific differences in expression of macrophage markers. Hepatic macrophages are labeled for CSF1R, Mac2 and F4/80, but lack CX3CR1 expression, whereas in the lung, the CSF1R+/Mac2+/Mac3+ macrophage population is not labeled with F4/80. In the splenic red pulp, subpopulations of CSF1R+/F4/80+/Mac3+cells were labeled with Mac2, CX3CR1 and lysozyme M. In the kidney, Mac2, Mac3 and lysozyme M labeled a fraction of the CSF1R+ and CX3CR1+ macrophages, but also stained tubular epithelial cells. In normal hearts, the majority of CSF1R+ and CX3CR1+ cells were not detected with anti-macrophage antibodies. Myocardial infarction was associated with marked expansion of the CSF1R+ and CX3CR1+ populations that peaked during the proliferative phase of cardiac repair, and also expressed Mac2, Mac3 and lysozyme M. In normal mouse tissues, a small fraction of cells labeled with anti-macrophage antibodies were identified as PDGFR $$\mathrm{\alpha }$$ α + fibroblasts, using a reporter system. The population of PDGFR $$\mathrm{\alpha }$$ α + cells expressing macrophage markers expanded following injury, likely reflecting emergence of cellular phenotypes with both fibroblast and macrophage characteristics. In conclusion, mouse macrophages exhibit remarkable heterogeneity. Selection of the most appropriate markers for identification of macrophages in mouse tissues is dependent on the organ and the pathologic condition studied.https://doi.org/10.1038/s41598-022-08278-x |
| spellingShingle | Bijun Chen Ruoshui Li Akihiko Kubota Linda Alex Nikolaos G. Frangogiannis Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies Scientific Reports |
| title | Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| title_full | Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| title_fullStr | Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| title_full_unstemmed | Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| title_short | Identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| title_sort | identification of macrophages in normal and injured mouse tissues using reporter lines and antibodies |
| url | https://doi.org/10.1038/s41598-022-08278-x |
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