XB-IMG-81710
Xenbase Image ID: 81710
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FIG. 2. Intracellular expression of the anti-ALK3 scFv D1 blocks ALK3 function in developing frog embryos as effectively as DN-ALK3. mRNA encoding the anti-ALK3 scFvs D1, E4, F2, DN-ALK3, the anti-FGFR1 scFv A2, the anti-Erb-B2 scFv F5, or the anti-cytochrome B scFv were injected into the two caudal cells (the so-called entralcells) of four-cell stage frog embryos. When the embryos reached the tadpole stage they were scored morphologically for trunk duplications characteristic of disrupting ALK3 receptor function. A: Expression of the anti-ALK3 scFv-D1 caused embryos to develop with trunk duplications. The data were compiled from three separate experiments and the number of injected embryos is indicated. B: Secondary trunks resulting from anti-ALK3 scFv-D1 expression are similar if not identical to those observed in embryos expressing the DN-ALK3. The trunk-duplications are indicated by the arrow. In addition to trunk duplications, a small number of the embryos expressing anti-ALK3 scFv-D1 or DN-ALK3 developed with enlarged heads, a phenotype also caused by disrupting ALK3 function (data not shown). C: Coexpression of wild-type ALK3 with anti-ALK3 scFv-D1 rescues the trunk duplications caused by anti-ALK3 scFv-D1 alone. D: Secondary trunks caused by anti-ALK3 scFv-D1 or the DN-ALK3 contained ectopic muscle (detected using whole-mount immunocytochemistry and the muscle-specific antibody 12/101) (Kintner and Brockes, 1984). The large arrowhead indicates the ectopic muscle. E: Anti-ALK3 scFv-D1 or the DN-ALK3 both directed the formation of secondary trunks that contained ectopic neural tubes (detected using whole-mount immunocytochemistry and the NCAM-specific 6F11 antibody). The large arrowhead indicates the ectopic neural tube that forms in the secondary trunks due to inhibition of ALK3 function. F: Expression of anti-ALK3 scFv-D1 or the DN-ALK3 caused ectopic expression of the myogenic factor myf-5. Stage 10.5 embryos expressing either anti-ALK3 scFv-D1 or the DN-ALK3 were analyzed for myf-5 expression using whole-mount in situ hybridization. i: Uninjected controls. ii: Embryos expressing DN-ALK3. iii: Embryos expressing Anti-ALK3 scFv-D1. iv: Embryos expressing anti-FGFR1 scFv-A2. Anti-ALK3 scFv-D1 and the DN-ALK3 caused ectopic myf-5 expression (marked by the arrow), while anti-FGFR1 scFv-A2 did not. The asterisk marks the early blastopore lip and Spemann organizer. Reproduced with permission of the Publisher, John Wiley & Sons. Image published in: Abler LL and Sheets MD (2003) Copyright © 2003. Image reproduced with permission of the Publisher, John Wiley & Sons.
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