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.
Journal of Cell Science   121(Pt 19): 3196-3206 (Journal)
Registered Authors
Crosier, Phil, Flores, Maria, Hall, Chris, Martin, Paul, Redd, Michael
WASP, Wound, Inflammation, Macrophage, Neutrophil, TILLING
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • Blood Coagulation/drug effects
  • Cell Movement/drug effects
  • Cell Survival/drug effects
  • Chemotaxis, Leukocyte/drug effects
  • Green Fluorescent Proteins/metabolism
  • Hematopoiesis/drug effects
  • Inflammation/metabolism*
  • Larva/drug effects
  • Larva/metabolism
  • Leukocytes/cytology
  • Leukocytes/drug effects
  • Macrophages/cytology
  • Macrophages/drug effects
  • Microscopy, Interference*
  • Molecular Sequence Data
  • Mutation/genetics
  • Neutrophils/cytology
  • Neutrophils/drug effects
  • Oligonucleotides, Antisense/pharmacology
  • Tail/pathology
  • Tail/ultrastructure
  • Time Factors
  • Wiskott-Aldrich Syndrome Protein/chemistry
  • Wiskott-Aldrich Syndrome Protein/metabolism*
  • Wounds and Injuries/metabolism*
  • Wounds and Injuries/pathology*
  • Zebrafish/embryology
  • Zebrafish/metabolism*
18782862 Full text @ J. Cell Sci.
Wiskott-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.
Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes