Analysis of In Vivo Cell Migration in Mosaic Zebrafish Embryos
Being optically clear, the zebrafish embryo is a nice model system to analyze cell migration in vivo. This chapter describes a combination of injection and cell transplant procedures that allows creation of mosaic embryos, containing a few cells labeled differently from their neighbors. Rapid 5D confocal imaging of these embryos permits to simultaneously track and quantify the movement of large cell groups, as well as analyze the cellular or subcellular dynamics of transplanted cells during their migration. In addition, expression of a candidate gene can be modified in transplanted cells. Comparing behavior of these cells to control or neighboring cells allows determination of the role of the candidate gene in cell migration. We describe the procedure, focusing on one specific cell population during gastrulation, but it can easily be adapted to other cell populations and other migration events during early embryogenesis.
Key wordsZebrafish Live imaging Cell migration Cell transplantation Mosaic embryos Cell tracking
This work was supported by the grant PJA 20151203256 from Fondation ARC pour la Recherche sur le Cancer, and grant ANR-15-CE13-0016-02 from Agence Nationale de la Recherche.
- 1.Horwitz R, Webb D (2003) Cell migration. Curr Biol 13:R756–R759. https://doi.org/10.1016/B978-0-12-394447-4.20070-9 CrossRefPubMedGoogle Scholar
- 4.te Boekhorst V, Preziosi L, Friedl P (2016) Plasticity of cell migration in vivo and in silico. Annu Rev Cell Dev Biol 3228361:1–28. https://doi.org/10.1146/annurev-cellbio-111315-125201 Google Scholar
- 11.Solnica-Krezel LL, Sepich DS (2012) Gastrulation: making and shaping germ layers. Annu Rev Cell Dev Biol 28:687–717. https://doi.org/10.1146/annurev-cellbio-092910-154043 CrossRefPubMedGoogle Scholar
- 17.Westerfield M (2000) The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), 4th edn. University of Oregon, Eugene, ORGoogle Scholar