PUBLICATION
Analysis of WASp function during the wound inflammatory response - live-imaging studies in zebrafish larvae
- Authors
- Cvejic, A., Hall, C., Bak-Maier, M., Flores, M.V., Crosier, P., Redd, M.J., and Martin, P.
- ID
- ZDB-PUB-080915-12
- Date
- 2008
- 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
- 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*
- PubMed
- 18782862 Full text @ J. Cell Sci.
Citation
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.
Abstract
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping