|ZFIN ID: ZDB-PUB-080915-12|
Analysis of WASp function during the wound inflammatory response - live-imaging studies in zebrafish larvae
Cvejic, A., Hall, C., Bak-Maier, M., Flores, M.V., Crosier, P., Redd, M.J., and Martin, P.
|Source:||Journal of Cell Science 121(Pt 19): 3196-3206 (Journal)|
|Registered Authors:||Crosier, Phil, Flores, Maria, Hall, Chris, Martin, Paul, Redd, Michael|
|Keywords:||WASP, Wound, Inflammation, Macrophage, Neutrophil, TILLING|
|PubMed:||18782862 Full text @ J. Cell Sci.|
Cvejic, A., Hall, C., Bak-Maier, M., Flores, M.V., Crosier, P., Redd, M.J., and Martin, P. (2008) Analysis of WASp function during the wound inflammatory response - live-imaging studies in zebrafish larvae. Journal of Cell Science. 121(Pt 19):3196-3206.
ABSTRACTWiskott-Aldrich syndrome protein (WASp) is haematopoietically restricted, and is the causative protein underlying a severe human disorder that can lead to death due to immunodeficiency and haemorrhaging. Much is known about the biochemistry of WASp and the migratory capacity of WASp-defective cells in vitro, but in vivo studies of immune-cell behaviour are more challenging. Using the translucency of zebrafish larvae, we live-imaged the effects of morpholino knockdown of WASp1 (also known as Was) on leukocyte migration in response to a wound. In embryos at 22 hours post-fertilisation, primitive macrophages were impaired in their migration towards laser wounds. Once a circulatory system had developed, at 3 days post-fertilisation, we observed significantly reduced recruitment of neutrophils and macrophages to ventral fin wounds. Cell-tracking studies indicated that fewer leukocytes leave the vessels adjacent to a wound and those that do exhibit impaired navigational capacity. Their cell morphology appears unaltered but their choice of leading-edge pseudopodia is more frequently incorrect, leading to impaired chemotaxis. We also identified two zebrafish mutants in WASp1 by TILLING, one of which was in the WIP-binding domain that is the hotspot for human lesions, and mutants exhibited the same deficiencies in wound inflammation and thrombus formation as WASp1 morphants.