Transplantation of GFP-expressing blastomeres for live imaging of retinal and brain development in chimeric zebrafish embryos.
Cells change extensively in their locations and property during embryogenesis. These changes are regulated by the interactions between the cells and their environment. Chimeric embryos, which are composed of cells of different genetic background, are great tools to study the cell-cell interactions mediated by genes of interest. The embryonic transparency of zebrafish at early developmental stages permits direct visualization of the morphogenesis of tissues and organs at the cellular level. Here, we demonstrate a protocol to generate chimeric retinas and brains in zebrafish embryos and to perform live imaging of the donor cells. The protocol covers the preparation of transplantation needles, the transplantation of GFP-expressing donor blastomeres to GFP-negative hosts, and the examination of donor cell behavior under live confocal microscopy. With slight modifications, this protocol can also be used to study the embryonic development of other tissues and organs in zebrafish. The advantages of using GFP to label donor cells are also discussed.
Zou
Jian
J
Department of Ophthalmology, University of Pittsburgh, PA, USA.
Wei
Xiangyun
X
D000818:Animals; D001757:Blastomeres / Q000378:metabolism / Q000637:transplantation*; D001921:Brain / Q000196:embryology*; D002678:Chimera; D004624:Embryo Transfer / Q000379:methods*; D005260:Female; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008297:Male; D018613:Microscopy, Confocal; D012160:Retina / Q000196:embryology*; D018183:Transplantation Chimera; D015027:Zebrafish / Q000196:embryology*
D049452:Green Fluorescent Proteins