XB-IMG-42251
Xenbase Image ID: 42251
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Fig. 8. Grafting experiments reveal a requirement for XtSulf1/2 in the environment through which cranial neural crest cells migrate. On the left, cartoons depict the design of the neural crest grafting experiments, while the panels to the right show the resulting embryos at tailbud stages. The top set of experiments show X. laevis embryos injected at the two-cell stage with 0.5 ng of GFP mRNA, ± 3 ng of mRNA coding for XtSulf2 and from which are dissected CNC explants at NF stage 14. Explants are transplanted into either a control embryo or an embryo injected with 3 ng of XtSulf2 mRNA. (A–C) Control host embryos transplanted at NF stage 14 with GFP expressing neural crest explants. (D–F) Host embryos injected with 3 ng of XtSulf2 at the two-cell stage into the animal pole and grafted with GFP expressing CNC explants at NF stage 14. (G–I) Control X. laevis host embryos grafted at NF stage 14 with GFP + XtSulf2 expressing CNC explants. All pictures were taken at 10X magnification; the eye, cement gland, and labelled cells are outlined. The bottom set of experiment show X. tropicalis embryos injected with mRNA coding for GFP ± AMOs targeted against XtSulf1 and XtSulf2. (J–L) At NF stage 15, CNC explants were taken from control Xenopus tropicalis embryos expressing GFP and transplanted into uninjected control embryos at the same stage. The GFP labelled cells migrate into the branchial arches by NF stage 30. (M–O) GFP mRNA together with AMOs targeting both XtSulf1 and XtSulf2 were injected into the animal hemisphere at the 2-cell stage and at NF stage 15 CNC explants were transplanted into control host embryos at the same stage. Labelled S1/S2 knockdown cells migrate less extensively than the control cells, but have direction. (P–R) At NF stage 15, CNC explants were taken from control Xenopus tropicalis embryos expressing GFP and transplanted into host embryos in which XtSulf1 and XtSulf2 have been knocked down. At stage 30, labelled cells are seen to be widely dispersed, suggesting that these wildtype cells lack direction in the S1/S2 mutant background. The eye, cement gland, and labelled cells are outlined. Image published in: Guiral EC et al. (2010) Copyright © 2010. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |