Measuring Inflammatory Cell Migration in the Zebrafish
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 wordsNeutrophil Migration Inflammation Zebrafish In vivo
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).
- 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.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.Nusslein-Volhard C, D.R. (2002) Zebrafish: A Practical Approach. Oxford University Press, Oxford.Google Scholar
- 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.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