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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 Fms intronic regulatory element (FIRE) within CSF1R is shown to be highly conserved in amniotes and absolutely required for myeloid-restricted expression of fluorescent reporter genes.The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein.In transgenic birds, expression of the reporter gene provided a defined marker for macrophage-lineage cells, identifying the earliest stages in the yolk sac, throughout embryonic development and in all adult tissues.

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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Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1+ B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1+ B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1+ B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1+ B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.
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DEV105593F5: Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1+ B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1+ B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1+ B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1+ B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.

Mentions: The expression of CSF1R-transgene expression in chicken tissue mononuclear subsets in the lymphoid organs and non-lymphoid tissues was examined by confocal microscopy. In the spleen, CSF1R-transgene-expressing cells were abundant and found in association with B-cells of the peri-ellipsoid lymphocyte sheath (PELS) and within the ellipsoid (Fig. 5A), consistent with previous studies of splenic macrophage populations (Jeurissen et al., 1989; Nagy et al., 2005; Igyártó et al., 2007). In the bursa of Fabricius, the avian-specific primary lymphoid organ for B-cell production, CSF1R-transgene-expressing cells were found in the medulla of B-cell follicles and in the interfollicular tissues (Fig. 5B). The location of CSF1R-transgene-expressing cells in the medulla is consistent with their identity as bursal secretory dendritic cells (BSDCs) (Oláh et al., 1992). Dense networks of CSF1R-transgene-expressing cells were present in the medulla region of germinal centres in the caecal tonsil (Fig. 5C). The distribution of cells in the medulla of germinal centres is consistent with cells previously described as avian follicular dendritic cells (FDCs) (Eikelenboom et al., 1983; Jeurissen, 1993). Both BSDCs and FDCs expressed high levels of CSF1R protein (supplementary material Fig. S4).Fig. 5.


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)

Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1+ B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1+ B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1+ B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1+ B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.
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Related In: Results  -  Collection

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DEV105593F5: Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1+ B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1+ B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1+ B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1+ B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.
Mentions: The expression of CSF1R-transgene expression in chicken tissue mononuclear subsets in the lymphoid organs and non-lymphoid tissues was examined by confocal microscopy. In the spleen, CSF1R-transgene-expressing cells were abundant and found in association with B-cells of the peri-ellipsoid lymphocyte sheath (PELS) and within the ellipsoid (Fig. 5A), consistent with previous studies of splenic macrophage populations (Jeurissen et al., 1989; Nagy et al., 2005; Igyártó et al., 2007). In the bursa of Fabricius, the avian-specific primary lymphoid organ for B-cell production, CSF1R-transgene-expressing cells were found in the medulla of B-cell follicles and in the interfollicular tissues (Fig. 5B). The location of CSF1R-transgene-expressing cells in the medulla is consistent with their identity as bursal secretory dendritic cells (BSDCs) (Oláh et al., 1992). Dense networks of CSF1R-transgene-expressing cells were present in the medulla region of germinal centres in the caecal tonsil (Fig. 5C). The distribution of cells in the medulla of germinal centres is consistent with cells previously described as avian follicular dendritic cells (FDCs) (Eikelenboom et al., 1983; Jeurissen, 1993). Both BSDCs and FDCs expressed high levels of CSF1R protein (supplementary material Fig. S4).Fig. 5.

Bottom Line: The Fms intronic regulatory element (FIRE) within CSF1R is shown to be highly conserved in amniotes and absolutely required for myeloid-restricted expression of fluorescent reporter genes.The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein.In transgenic birds, expression of the reporter gene provided a defined marker for macrophage-lineage cells, identifying the earliest stages in the yolk sac, throughout embryonic development and in all adult tissues.

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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Related in: MedlinePlus