XB-IMG-43115
Xenbase Image ID: 43115
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Figure 1. FN Antisense Morpholino “Knocks down” FN Protein Synthesis, Reduces Fibril Formation, and Reduces Dorsal Axis Extension(A) Western blot analysis of morpholino (MO)-injected embryos harvested at stage 10.5 and processed for detection with the anti-FN mAb 4H2 and the anti-integrin β1 PcAb 363. The two FN bands correspond to two major alternatively spliced FN subunits expressed during development. The control MO has no affect on the levels of expression of either FN or integrin β1 at up to 15 μMoles MO injected per embryo. A dose-dependent reduction in FN protein expression is noted in the presence of the combined (50:50) FNMOs. Levels of integrin β1 protein expression are unaffected by the FN antisense MO.(B) Expression of chordin, a marker of dorsal axial tissues, shows that embryos injected with 15 μMoles FNMO have wide dorsal axial tissues. (Arrows mark the width of the chordin expression.)(C) Transverse confocal sections of embryos stained for FN fibrils show a typical pattern in both control morpholino and uninjected control embryos while fibril formation is almost completely abolished (see arrowheads for an example of faint labeling of blastocoel roof ectoderm in FNMO-injected embryos).(D) FNMO-injected embryos show defects in both dorsal and ventral elongation by early tadpole stages and severe anterior mesoderm defects by later tadpole stages. At early tadpole stages, the dorsal axial extension is moderately reduced while ventral extension is severely reduced (asterisk). Late-stage FNMO-injected tadpoles show defects in ventral mesoderm morphogenesis, ectodermal lesions, and failure of heart (arrow) and gut (arrowheads) formation.(E) Pooled data from three batches of embryos were observed over several days and their progress was assessed. None of the embryos injected with 15 μMoles FNMO develop beating hearts. Image published in: Davidson LA et al. (2006) Copyright © 2006. Image reproduced with permission of the Publisher, Elsevier B. V.
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