Howard Hughes Medical Institute, University of California, Los Angeles, CA 90095-1662.
Department of Biological Chemistry, University of California, Los Angeles, CA 90095-1662.
Proc Natl Acad Sci U S A. 2017 Apr 11;114(15):E3081-E3090. doi: 10.1073/pnas.1700766114. Epub 2017 Mar 27.
The earliest event in development is the dorsal accumulation of nuclear β-catenin under the influence of cytoplasmic determinants displaced by fertilization. In this study, a genome-wide approach was used to examine transcription of the 43,673 genes annotated in the genome under a variety of conditions that inhibit or promote formation of the Spemann organizer signaling center. Loss of function of β-catenin with antisense morpholinos reproducibly reduced the expression of 247 mRNAs at gastrula stage. Interestingly, only 123 β-catenin targets were enriched on the dorsal side and defined an early dorsal β-catenin gene signature. These genes included several previously unrecognized Spemann organizer components. Surprisingly, only 3 of these 123 genes overlapped with the late Wnt signature recently defined by two other groups using inhibition by mRNA or Wnt8 morpholinos, which indicates that the effects of β-catenin/Wnt signaling in early development are exquisitely regulated by stage-dependent mechanisms. We analyzed transcriptome responses to a number of treatments in a total of 46 RNA-seq libraries. These treatments included, in addition to β-catenin depletion, regenerating dorsal and ventral half-embryos, lithium chloride treatment, and the overexpression of , , and mRNAs. Only some of the early dorsal β-catenin signature genes were activated at blastula whereas others required the induction of endomesoderm, as indicated by their inhibition by Cerberus overexpression. These comprehensive data provide a rich resource for analyzing how the dorsal and ventral regions of the embryo communicate with each other in a self-organizing vertebrate model embryo.
发育过程中的最早事件是在细胞质决定因素被受精所取代的影响下,β-连环蛋白在背部的积累。在这项研究中,我们采用了一种全基因组的方法,在各种抑制或促进 Spemann 组织者信号中心形成的条件下,检查了 43673 个注释基因的转录。用反义 morpholino 使 β-连环蛋白失活,可重复性地降低了原肠胚期 247 个 mRNA 的表达。有趣的是,只有 123 个 β-连环蛋白靶点在背部富集,并定义了一个早期的背部 β-连环蛋白基因特征。这些基因包括几个以前未被识别的 Spemann 组织者成分。令人惊讶的是,这 123 个基因中只有 3 个与最近另外两个使用抑制 mRNA 或 Wnt8 morpholino 的小组定义的晚期 Wnt 特征重叠,这表明β-连环蛋白/Wnt 信号在早期发育中的作用受到阶段依赖性机制的精细调控。我们分析了总共 46 个 RNA-seq 文库中许多处理的转录组反应。除了β-连环蛋白耗竭之外,这些处理还包括再生背腹半胚胎、氯化锂处理以及 、 和 mRNA 的过表达。只有一些早期的背部β-连环蛋白特征基因在原肠胚期被激活,而其他基因则需要诱导内胚层和中胚层,这可以通过 Cerberus 过表达抑制它们来证明。这些全面的数据为分析胚胎背腹区域如何在自我组织的脊椎动物模型胚胎中相互通信提供了丰富的资源。