PUBLICATION

An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo

Authors
Jiang, Y., Chen, J., Xu, J.
ID
ZDB-PUB-191015-15
Date
2019
Source
Journal of visualized experiments : JoVE   (151): (Journal)
Registered Authors
Chen, Jiahao, Jiang, Yunyun
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Cell Movement
  • Chemokines/metabolism*
  • Embryo, Nonmammalian/metabolism
  • Macrophages/cytology
  • Macrophages/metabolism*
  • Models, Biological*
  • Software
  • Zebrafish/embryology
  • Zebrafish/genetics
PubMed
31609330 Full text @ J. Vis. Exp.
Abstract
Zebrafish are widely used in basic and biomedical research. Many zebrafish transgenic lines are currently available to label various types of cells. Owing to the transparent embryonic body of zebrafish, it is convenient for us to study the effect of one chemokine on the behavior of a certain type of cells in vivo. Here we provided a workflow to investigate the function of a chemokine on macrophage migration in vivo. We constructed a tissue-specific overexpression plasmid to overexpress IL-34 and injected the plasmid into one-cell stage transgenic fish embryos whose macrophages were specifically labeled by a fluorescent protein. We then used whole mount fluorescent in situ hybridization and immunostaining to detect the pattern of the chemokine expression and the number or location of macrophages. The injected WT embryos were raised to generate a stable transgenic line. Finally, we used confocal live imaging to directly observe macrophage behavior in the stable transgenic fish to study the function of IL-34 on macrophages in vivo.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping