PUBLICATION

Modelling human Wiskott-Aldrich syndrome protein mutants in zebrafish larvae using live in vivo imaging

Authors
Jones, R.A., Feng, Y., Worth, A.J., Thrasher, A.J., Burns, S.O., and Martin, P.
ID
ZDB-PUB-130805-9
Date
2013
Source
Journal of Cell Science   126(Pt 18): 4077-84 (Journal)
Registered Authors
Feng, Yi, Jones, Rebecca Amy, Martin, Paul
Keywords
Live imaging, Disease model, WASP, Zebrafish, Immune deficiency, Neutrophils, Macrophages
MeSH Terms
  • Neutropenia/congenital*
  • Neutropenia/genetics
  • Neutropenia/metabolism
  • Macrophages/metabolism*
  • Disease Models, Animal
  • Zebrafish
  • Wiskott-Aldrich Syndrome/genetics*
  • Wiskott-Aldrich Syndrome/metabolism
  • Humans
  • Animals
(all 10)
PubMed
23868979 Full text @ J. Cell Sci.
Abstract

Wiskott Aldrich syndrome (WAS) and X-linked neutropenia (XLN) are immunodeficiencies in which the functions of several haematopoietic cell lineages are perturbed due to mutations in the actin regulator WASp. From in vitro cell biology experiments and biochemical and structural approaches we know much about the functional domains of WASp, and how WASp might regulate the dynamic actin cytoskeleton downstream of activators such as Cdc42, but in vivo experiments are much more challenging. In patients there is a correlation between clinical disease and genotype, with severe reductions in WASp expression or function associating with complex multilineage immunodeficiency, whereas, specific mutations that cause constitutive activation of WASp result in congenital neutropenia. Here we take advantage of the genetic tractability and translucency of zebrafish larvae to first characterise how a null mutant in zfWASp influences the behaviour of neutrophils and macrophages in response to tissue damage and to clearance of infections. We then use this mutant background to study how leukocyte lineage-specific transgenic replacement with human WASp variants, (including normal wild type, and point mutations that either fail to bind Cdc42 or cannot be phosphorylated, and a constitutively active mutant equivalent to that seen in XLN patients), alter the capacity for generation of neutrophils, and their chemotactic response to wounds, and the phagocytic clearance capacity of macrophages. This model provides a unique insight into WASp-related immunodeficiency at both a cellular and whole organism level.

Genes / Markers
Figures
Figure Gallery (5 images)
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
bsl251TgTransgenic Insertion
    bsl252TgTransgenic Insertion
      bsl253TgTransgenic Insertion
        bsl254TgTransgenic Insertion
          bsl255TgTransgenic Insertion
            hu3280
              Point Mutation
              i114TgTransgenic Insertion
                i149TgTransgenic Insertion
                  i186TgTransgenic Insertion
                    i252bTgTransgenic Insertion
                      1 - 10 of 13
                      Show
                      Human Disease / Model
                      Human Disease Fish Conditions Evidence
                      Wiskott-Aldrich syndromewasbhu3280/hu3280standard conditionsTAS
                      1 - 1 of 1
                      Show
                      Sequence Targeting Reagents
                      Target Reagent Reagent Type
                      wasaMO1-wasaMRPHLNO
                      wasaMO2-wasaMRPHLNO
                      1 - 2 of 2
                      Show
                      Fish
                      Antibodies
                      Name Type Antigen Genes Isotypes Host Organism
                      Ab1-wasbpolyclonal
                        Rabbit
                        Ab2-lcp1polyclonalRabbit
                        1 - 2 of 2
                        Show
                        Orthology
                        No data available
                        Engineered Foreign Genes
                        Marker Marker Type Name
                        DsRed2EFGDsRed2
                        EGFPEFGEGFP
                        GAL4EFGGAL4
                        GFPEFGGFP
                        KaedeEFGKaede
                        mCherryEFGmCherry
                        NTREFGNTR
                        1 - 7 of 7
                        Show
                        Mapping
                        No data available