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Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage.

Balic A, Garcia-Morales C, Vervelde L, Gilhooley H, Sherman A, Garceau V, Gutowska MW, Burt DW, Kaiser P, Hume DA, Sang HM - Development (2014)

Bottom Line: The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein.The reporter genes permit detailed and dynamic visualisation of embryonic chicken macrophages.Chicken embryonic macrophages are not recruited to incisional wounds, but are able to recognise and phagocytose microbial antigens.

View Article: PubMed Central - PubMed

Affiliation: The Roslin Institute and Royal (Dick) School of Veterinary Sciences, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK adam.balic@roslin.ed.ac.uk helen.sang@roslin.ed.ac.uk.

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F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R-mApple-expressing cells (red) in relation to: (J) Bu-1+ B-cells (green), (K) TCR αβ (Vβ1)+ T-cells (green) and (L) CVI-ChNL-74.2+ macrophages (green). Scale bars in J-L: 100 µm.
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DEV105593F6: F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R-mApple-expressing cells (red) in relation to: (J) Bu-1+ B-cells (green), (K) TCR αβ (Vβ1)+ T-cells (green) and (L) CVI-ChNL-74.2+ macrophages (green). Scale bars in J-L: 100 µm.

Mentions: We observed CSF1R-transgene-expressing cells in the brain (Fig. 5D). Their CD45+ phenotype and highly ramified appearance is consistent with their identity as microglial cells (Cuadros et al., 2006), the resident macrophage population of neuronal tissues. Similarly, macrophages of the liver (Kupffer cells) were located in the sinusoids, as expected (Fig. 5E). In contrast to mammalian lung, the avian lung does not contain alveoli or cells equivalent to alveolar macrophages, but there is a network of phagocytes surrounding the larger airways (de Geus et al., 2012). Consistent with this pattern, CSF1R-transgene-expressing cells were scattered throughout the interstitial tissue of the parabronchial wall and clustered with B-cells to form small, isolated lymphoid follicles in the lung (Fig. 5F). Epidermal sheet preparations contained large numbers of transgene-expressing cells, both scattered cells and in small clusters (Fig. 5G), consistent with reported distribution of chicken Langerhans cells (Igyártó et al., 2006). Unexpectedly, in the skeletal muscle we observed many CSF1R-transgene-expressing cells. These cells co-expressed class II MHC (Fig. 6H) and were also positive for CSF1R (not shown), indicating they are resident skeletal muscle macrophages. One other macrophage population that is unique to birds is in the skin, where the transgene highlighted the major haematopoietic cell subset in feather pulp (Fig. 5I).Fig. 6.


Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage.

Balic A, Garcia-Morales C, Vervelde L, Gilhooley H, Sherman A, Garceau V, Gutowska MW, Burt DW, Kaiser P, Hume DA, Sang HM - Development (2014)

F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R-mApple-expressing cells (red) in relation to: (J) Bu-1+ B-cells (green), (K) TCR αβ (Vβ1)+ T-cells (green) and (L) CVI-ChNL-74.2+ macrophages (green). Scale bars in J-L: 100 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4197536&req=5

DEV105593F6: F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R-mApple-expressing cells (red) in relation to: (J) Bu-1+ B-cells (green), (K) TCR αβ (Vβ1)+ T-cells (green) and (L) CVI-ChNL-74.2+ macrophages (green). Scale bars in J-L: 100 µm.
Mentions: We observed CSF1R-transgene-expressing cells in the brain (Fig. 5D). Their CD45+ phenotype and highly ramified appearance is consistent with their identity as microglial cells (Cuadros et al., 2006), the resident macrophage population of neuronal tissues. Similarly, macrophages of the liver (Kupffer cells) were located in the sinusoids, as expected (Fig. 5E). In contrast to mammalian lung, the avian lung does not contain alveoli or cells equivalent to alveolar macrophages, but there is a network of phagocytes surrounding the larger airways (de Geus et al., 2012). Consistent with this pattern, CSF1R-transgene-expressing cells were scattered throughout the interstitial tissue of the parabronchial wall and clustered with B-cells to form small, isolated lymphoid follicles in the lung (Fig. 5F). Epidermal sheet preparations contained large numbers of transgene-expressing cells, both scattered cells and in small clusters (Fig. 5G), consistent with reported distribution of chicken Langerhans cells (Igyártó et al., 2006). Unexpectedly, in the skeletal muscle we observed many CSF1R-transgene-expressing cells. These cells co-expressed class II MHC (Fig. 6H) and were also positive for CSF1R (not shown), indicating they are resident skeletal muscle macrophages. One other macrophage population that is unique to birds is in the skin, where the transgene highlighted the major haematopoietic cell subset in feather pulp (Fig. 5I).Fig. 6.

Bottom Line: The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein.The reporter genes permit detailed and dynamic visualisation of embryonic chicken macrophages.Chicken embryonic macrophages are not recruited to incisional wounds, but are able to recognise and phagocytose microbial antigens.

View Article: PubMed Central - PubMed

Affiliation: The Roslin Institute and Royal (Dick) School of Veterinary Sciences, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK adam.balic@roslin.ed.ac.uk helen.sang@roslin.ed.ac.uk.

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