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Distribution of macrophages during fish development: an immunohistochemical study in carp (Cyprinus carpio, L.)
N Romano1, S Picchietti, J J Taverne-Thiele
1Anatomia Comparata, Dipartimento di Scienze Ambientali, Blocco D, Università della Tuscia, Viterbo, Italy. nromano@unitus.it
Insights
A new antibody (WCL15) tracks carp macrophages in developing lymphoid tissues. These immune cells appear early and play a role in fish defense and immune cell development.
Area of Science:
- Immunology
- Fish Biology
- Cell Biology
Background:
- Macrophages are crucial immune cells in vertebrates.
- Understanding macrophage distribution in fish is vital for aquaculture and disease resistance.
- Carp (Cyprinus carpio) are important aquaculture species, but their immune system development requires further study.
Purpose of the Study:
- To develop and utilize a monoclonal antibody (WCL15) to map the distribution of macrophages in developing carp lymphoid tissues.
- To investigate the temporal and spatial emergence of macrophages during early carp development.
- To discuss the functional implications of macrophage presence in early carp immunity.
Main Methods:
- Development of a monoclonal antibody (WCL15) targeting carp macrophages.
- Application of flow cytometry, immuno-histochemistry, and immuno-electron microscopy for cell analysis.
- Double immunostaining to differentiate macrophages from thrombocytes.
Main Results:
- The WCL15 antibody effectively identified macrophages in both live cells and fixed tissues.
- Macrophages were detected as early as day 2 post-fertilization in head kidney and yolk sac epithelium.
- Macrophages were subsequently found in thymus, gut, and spleen, with numbers peaking at 6-8 weeks before redistributing.
Conclusions:
- The WCL15 antibody is a valuable tool for studying carp macrophage development and distribution.
- Macrophages are present early in carp development, suggesting a role in innate immunity.
- The localization of macrophages in specific lymphoid areas indicates their involvement in immune cell differentiation and tissue-specific immune responses.
Abstract:
A monoclonal antibody against carp macrophages (WCL15) has been utilised in flow cytometry, immuno-histochemistry and immuno-electron microscopy to assess the distribution of monocytes/macrophages in developing carp lymphoid tissues. In suspensions of living cells WCL15 reacted strongly with cytoplasm and plasmic membrane of macrophages. It also cross-reacted with a subpopulation of thrombocytes, but this reaction could be neglected by double immunostaining in combination with a thrombocyte-specific marker. In Bouin-fixed tissues the antibody distinctly recognised macrophages. Macrophages were found from day 2 post-fertilisation in head kidney and in the dorsal portion of the yolk sac epithelium. From 1 week onwards macrophages were found scattered in thymus and gut and during the second week in spleen. Macrophages increased in number in all lymphoid tissues until the 6-8th week post-fertilisation, but they decreased except in thymus, where they became localised mainly in the cortical-medullary boundary, and in white pulp areas of head kidney. The role of macrophages in allowing an early non-specific defence in young fish and in co-operating during the differentiation processes of T-cells and B-cells is discussed.
