Maternal Wnt11b regulates cortical rotation during Xenopus axis formation: analysis of maternal-effect wnt11b mutants.

Maternal Wnt11b regulates cortical rotation during Xenopus axis formation: analysis of maternal-effect wnt11b mutants.
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DOI:
10.1242/dev.200552
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发表时间:
2022-09-01
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Development (Cambridge, England)
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早期胚胎中的不对称信号中心对于脊椎动物轴的形成至关重要。这些区域(例如两栖动物背侧桑葚胚、哺乳动物前内脏内胚层)需要稳定的核 β-连环蛋白,但局部 Wnt 配体信号传导活性在其建立中的作用仍不清楚。在非洲爪蟾中,背侧β-连环蛋白是由受精卵中植物微管介导的对称性破坏启动的,称为“皮质旋转”。局部化的 wnt11b mRNA 和 β-catenin 的配体独立激活剂与背侧 β-catenin 激活有关,但在这种范式中,每种激活剂对轴形成的贡献程度仍不清楚。在这里,我们描述了非洲爪蟾 wnt11b.L 中 CRISPR 介导的母体效应突变。我们发现,wnt11b 对于强健的背轴形成和及时的原肠胚形成是母体必需的,对于左右不对称也是合子必需的。重要的是,我们发现 wnt11b 突变卵中植物微管组装和皮质旋转减少。此外,我们发现激活的 Wnt 辅助受体 Lrp6 和 Disheveled 缺乏与早期 β-catenin 稳定作用一致的行为,并且两者都不受 Wnt11b 调节。因此,这项工作表明 Wnt11b 参与了假定的背部决定簇的分布,而不是决定簇本身的组成。摘要:母体 Wnt11b 在皮质旋转过程中调节微管动力学。
Asymmetric signalling centres in the early embryo are essential for axis formation in vertebrates. These regions (e.g. amphibian dorsal morula, mammalian anterior visceral endoderm) require stabilised nuclear β-catenin, but the role of localised Wnt ligand signalling activity in their establishment remains unclear. In Xenopus, dorsal β-catenin is initiated by vegetal microtubule-mediated symmetry breaking in the fertilised egg, known as ‘cortical rotation’. Localised wnt11b mRNA and ligand-independent activators of β-catenin have been implicated in dorsal β-catenin activation, but the extent to which each contributes to axis formation in this paradigm remains unclear. Here, we describe a CRISPR-mediated maternal-effect mutation in Xenopus laevis wnt11b.L. We find that wnt11b is maternally required for robust dorsal axis formation and for timely gastrulation, and zygotically for left-right asymmetry. Importantly, we show that vegetal microtubule assembly and cortical rotation are reduced in wnt11b mutant eggs. In addition, we show that activated Wnt coreceptor Lrp6 and Dishevelled lack behaviour consistent with roles in early β-catenin stabilisation, and that neither is regulated by Wnt11b. This work thus implicates Wnt11b in the distribution of putative dorsal determinants rather than in comprising the determinants themselves. Summary: Maternal Wnt11b regulates microtubule dynamics during cortical rotation.
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