β-catenin signaling activity dissected in the early Xenopus embryo:: A novel antisense approach
β-catenin signaling activity dissected in the early Xenopus embryo:: A novel antisense approach
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DOI：
10.1006/dbio.2000.9720
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发表时间：
2000-06-01
影响因子：
2.7
通讯作者：
Wylie, C
中科院分区：
文献类型：
--
作者：
Heasman, J;Kofron, M;Wylie, C
非洲爪蟾胚胎发育的背/腹和前/后轴作为一个母亲的Xwnt途径，其中β-连环蛋白是一个必不可少的组成部分，作为合子基因转录的反式激活因子的活动的结果。然而，β-连环蛋白在早期胚胎发生中何时何地需要的问题还没有直接解决，因为没有功能丧失的方法。在这里，我们报告使用一种新的反义方法，使我们能够针对个别卵裂球的蛋白质消耗。当将与β-连环蛋白mRNA互补的“吗啉代”寡核苷酸注射到早期胚胎中时，它通过神经胚阶段有效地消耗β-连环蛋白。通过将寡核苷酸靶向不同的卵裂球，我们在不同的区域和不同的时间阻断了β-连环蛋白的活性。在2-和4-细胞阶段的背侧植物注射阻断背轴形成，在8-细胞阶段阻断头部形成，而在32-细胞阶段的A-层注射导致异常的水泥腺形成。这种方法显示了复杂的参与Xwnt途径在胚胎图案，并提供了一个快速的方法，在非洲爪蟾母源和早期合子基因产物的功能分析。(C)北京大学出版社.
Xenopus embryos develop dorsal/ventral and anterior/posterior axes as a result of the activity of a maternal Xwnt pathway, in which beta-catenin is an essential component, acting as a transactivator of transcription of zygotic genes. However, the questions of where and when beta-catenin is required in early embryogenesis have not been addressed directly, because no loss-of-function method has been available. Here we report the use of a novel antisense approach that allows us to target depletion of protein to individual blastomeres. When a "morpholino" oligo complementary to beta-catenin mRNA is injected into early embryos, it depletes beta-catenin protein effectively through the neurula stage. By targeting the oligo to different cleavage blastomeres, we block beta-catenin activity in different areas and at different times. Dorsal vegetal injection at the 2- and 4-cell stages blocks dorsal axis formation and at the 8-cell stage blocks head formation, while A-tier injection at the 32-cell stage causes abnormal cement gland formation. This approach shows the complex involvement of Xwnt pathways in embryonic patterning and offers a rapid method for the functional analysis of both maternal and early zygotic gene products in Xenopus. (C) 2000 Academic Press.