Figure 1. The Metalloproteinase Activity of ADAM13 Is Required for Xenopus CNC Induction(A) cDNA sequence of adam13 showing the translation start site (red) and targets of MOs 13-1 and 13-3.(B) One blastomere of two-cell stage embryos was injected with MO 13-1 (6 ng) and transcripts encoding myc-tagged ADAMs (ADAM12 or 13: 200 pg; ADAM19: 100 pg). Embryos were cultured for 20 hr and western blot of whole-embryo lysates carried out using an anti-myc antibody.(C) Western blot of stage 11–12 embryo lysates showing KD of endogenous ADAM13 by injection of MO 13-1 at one-cell stage (12 ng MO/embryo; 20 embryos/lane).(D) Head cartilage phenotypes of ADAM13 KD. One anterior-dorsal blastomere of eight-cell stage embryos was injected with the indicated MO (1.5 ng). Embryos were allowed to develop to stage 46 and scored for head cartilage defects. Images were gray-scale inverted for easier visualization of head cartilage. One example of each phenotype is shown in the upper panels, and results of three independent experiments are graphed in the lower panel (error bars represent SDs). *p = 0.03; **p = 0.005; NS, not significant (p = 0.43). See Figure S1D and Experimental Procedures for details on phenotype scoring and statistics.(E and F) Four-cell stage embryos were injected in one anterior-dorsal blastomere with 3 ng MO 13-3, allowed to develop to stage 12.5/13, and then processed by in situ hybridization for snail2 (E) or sox9 (F).(G) Embryos were injected with the indicated MO (3 ng) and “rescue” transcripts (100 pg each), and in situ hybridization performed as in (E). Red asterisks in (D)–(G) denote injected side.Single and double arrowheads in (B) and (C) point to the pro- and mature forms of ADAMs, respectively, and arrows in (D) point to impaired head cartilage structures as compared with the uninjected side. U, uninjected; CT, control MO; n, number of embryos scored (same below).
Image published in: Wei S et al. (2010)
Copyright © 2010. Image reproduced with permission of the Publisher, Elsevier B. V.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| snai2.L | slug, snai2-a, snai2-b, Snail2, xSlu, xslug, XSnail2 | X. laevis | Sometime during NF stage 12.5 to NF stage 13 | neural plate cranial neural crest |
| sox9.S | cmd1, cmpd1, sox-9, sox9-a, sox9-b, sra1 | X. laevis | Sometime during NF stage 12.5 to NF stage 13 | neural plate |
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