Fig. 7

plcβ3 is required in cranial neural crest cells for pharyngeal arch patterning. (A--C) Projections of confocal z-stacks, including fluorescent and bright-field channels, of lateral and orthogonal views of live 36 hpf mosaic host embryos. (A) Cells from the animal pole of Alexa568-labeled sphere staged (6 hpf) wild-type fli:GFP donors were transplanted unilaterally into (A) the neural crest domain, (B) the surface ectoderm domain, and (C) the endoderm domain of plcβ3 homozygous hosts. Embryos were screened at 30–36 hpf to determine contribution of donor cells to pharyngeal arches and again at 6 dpf to assess contribution of donor cells to cartilages. Dots indicate bottom of the first pharyngeal pouch. Green florescence indicates presence of fli-positive donor neural crest in transplants in panel A and absence of green florescence indicates absence of neural crest in transplants in panels B and C. (D--F) Embryos imaged in panels A–C were subsequently fixed and doubly stained for both cartilage (Alcian blue) and bone (Alizarin red). Non-transplant sides of hosts were used as negative controls for rescue. As brain tissues frequently accompany large neural crest transplants, we also analyzed transplants containing only brain tissue and determined they did not rescue (n = 5, data not shown). Scale bars: 50 μm.