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FIG. 9. Localization of X-ADAM 13 protein during development. Whole-mount embryos were stained with the affinity-purified cytoplasmic tail antibody and alkaline phosphatase-conjugated secondary antibody. (A) Lateral view of an early tailbud (stage 22, top) and late neurula (stage 19, bottom) stage embryo. The staining is consistent with mRNA localization in the somites (red arrowhead) and in the cranial neural crest (black arrows and arrowhead. (B) Magnification of the anterior portion of a tailbud (stage 22) embryo. The somitic staining is strongest at the intersomitic border (red arrowheads). In the most anterior somites, the entire cell surface is stained. In the head area, staining is localized to the four cranial neural crest-derived segments (black arrows and arrowheads): posterior branchial crest (p), anterior branchial crest (a), hyoid crest (h), and mandibular crest segment (m). Staining of this last segment (m) completely surrounds the optic vesicle at this stage (black arrowhead). (C) Section of tailbud stage embryo triple-labeled with DATF-conjugated anti-rabbit secondary antibody, Texas-red-conjugated anti-mouse secondary antibody and DAPI. The X-ADAM 13 protein appears in green, fibronectin appears in red, and DAPI-labeled nuclei appear in blue. In this frontal section that passes through the somites and notochord, X-ADAM 13 protein is localized to individual myocyte end junctions. The fibronectin staining is present in the extracellular space between somites (appears yellow due to overlap with green X- ADAM 13 staining) as well as the periphery of the notochord (stained red). Nuclear staining indicates the organization of somitic cells. Primary antibodies are 6615 (anti-X-ADAM 13) and 4H2 (anti-fibronectin: Ramos and DeSimone, 1996). |
