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Cell Migration pp 261-275 | Cite as

Measuring Inflammatory Cell Migration in the Zebrafish

  • Philip M. Elks
  • Catherine A. Loynes
  • Stephen A. Renshaw
Part of the Methods in Molecular Biology book series (MIMB, volume 769)

Abstract

A key feature of inflammatory cells is the ability to migrate to a site of injury or infection quickly and efficiently. Infectious agents can then be taken up by these inflammatory cells, preventing established infection. Inflammatory cell migration is driven by a complex interaction between inflammatory cells and their environment. In order to maintain health, inflammation needs to resolve, allowing the surrounding tissues to recover and heal. These processes are not fully understood and have been difficult to study in cell culture due to the complex interactions between cell types. We therefore use a range of techniques in near-transparent zebrafish (Danio rerio) larvae to study these migration events in a whole-organism, in vivo model. Using a transgenic zebrafish line that specifically marks neutrophils with green fluorescent protein, Tg(mpx:GFP)i114, we are able to follow neutrophil behaviour at a single cell level. Using these methods, the cellular processes involved in all phases of inflammation can be studied and better understood.

Key words

Neutrophil Migration Inflammation Zebrafish In vivo 

Notes

Acknowledgements

SAR and CAL are funded by an MRC Senior Clinical Fellowship to SAR (reference number: G0701932). PME is funded by a project grant from the Wellcome Trust (reference number: WT082909MA). Microscopy studies in our laboratory are supported by a Wellcome Trust grant to the MBB/BMS Light microscopy facility (GR077544AIA), and the laboratory is supported by an MRC Centre grant (G0700091).

References

  1. 1.
    Lieschke, G.J., Oates, A.C., Crowhurst, M.O., Ward, A.C., Layton, J.E. (2001) Morphologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish. Blood 98, 3087–96.PubMedCrossRefGoogle Scholar
  2. 2.
    Lieschke, G.J., Currie, P.D. (2007) Animal models of human disease: zebrafish swim into view. Nat Rev Genet 8, 353–67.PubMedCrossRefGoogle Scholar
  3. 3.
    Trede, N.S., Langenau, D.M., Traver, D., Look, A.T., Zon, L.I. (2004) The use of zebrafish to understand immunity. Immunity 20, 367–79.PubMedCrossRefGoogle Scholar
  4. 4.
    Carradice, D., Lieschke, G.J. (2008) Zebrafish in hematology: sushi or science? Blood 111, 3331–42.PubMedCrossRefGoogle Scholar
  5. 5.
    Renshaw, S.A., Loynes, C.A., Trushell, D.M., Elworthy, S., Ingham, P.W., Whyte, M.K. (2006) A transgenic zebrafish model of neutrophilic inflammation. Blood 108, 3976–8.PubMedCrossRefGoogle Scholar
  6. 6.
    Nasevicius, A., Ekker, S.C. (2000) Effective targeted gene ‘knockdown’ in zebrafish. Nat Genet 26, 216–20.PubMedCrossRefGoogle Scholar
  7. 7.
    Savill, J.S., Wyllie, A.H., Henson, J.E., Walport, M.J., Henson, P.M., Haslett, C. (1989) Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutrophil leads to its recognition by macrophages. J Clin Invest 83, 865–75.PubMedCrossRefGoogle Scholar
  8. 8.
    Martin, R.T., Bartman, T. (2009) Analysis of heart valve development in larval zebrafish. Dev Dyn 238, 1796–802.PubMedCrossRefGoogle Scholar
  9. 9.
    Brown, S.B., Tucker, C.S., Ford, C., Lee, Y., Dunbar, D.R., Mullins, J.J. (2007) Class III antiarrhythmic methanesulfonanilides inhibit leukocyte recruitment in zebrafish. J Leukoc Biol 82, 79–84.PubMedCrossRefGoogle Scholar
  10. 10.
    Redd, M.J., Cooper, L., Wood, W., Stramer, B., Martin, P. (2004) Wound healing and inflammation: embryos reveal the way to perfect repair. Philos Trans R Soc Lond B Biol Sci 359, 777–84.PubMedCrossRefGoogle Scholar
  11. 11.
    Herbomel, P., Thisse, B., Thisse, C. (2001) Zebrafish early macrophages colonize cephalic mesenchyme and developing brain, retina, and epidermis through a M-CSF receptor-dependent invasive process. Dev Biol 238, 274–88.PubMedCrossRefGoogle Scholar
  12. 12.
    Loynes, C.A., Martin, J.S., Robertson, A., Trushell, D.M., Ingham, P.W., Whyte, M.K., Renshaw, S.A. Pivotal Advance: Pharmaco­logical manipulation of inflammation resolution during spontaneously resolving tissue neutrophilia in the zebrafish. J Leukoc Biol 87, 203–12.Google Scholar
  13. 13.
    Elks, P.M., van Eeden, F.J., Dixon, G., Wang, X., Reyes-Aldasoro, C.C., Ingham, P.W., Whyte, M.K.B., Walmsley, S.R., and Renshaw, S.A. (2011) Activation of Hif-1alpha delays inflammation resolution by reducing neutrophil apoptosis and reverse migration in a zebrafish inflammation model. Blood. In press.Google Scholar
  14. 14.
    Nusslein-Volhard C, D.R. (2002) Zebrafish: A Practical Approach. Oxford University Press, Oxford.Google Scholar
  15. 15.
    Ellett, F., Pase, L., Hayman, J.W., Andrian-opoulos, A., and Lieschke, G.J. (2010) mpeg1 promoter transgenes direct macrophage-lineage expression in zebrafish. Blood. In press.Google Scholar
  16. 16.
    Gray, C., Loynes, C.A., Whyte, M.K.B., Crossman, D.C., Renshaw, S.A., and Chico, T.J.A. (2011) Simultaneous intravital imaging of macrophage and neutrophil behaviour during inflammation using a novel transgenic zebrafish. Thrombosis and Haemostasis 105.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Philip M. Elks
  • Catherine A. Loynes
  • Stephen A. Renshaw
    • 1
  1. 1.MRC Centre for Developmental and Biomedical GeneticsThe University of SheffieldSheffieldUK

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