Gene Expression in Amphibian Development

项目总结/摘要 我们的目标是在分子水平上了解脊椎动物的早期发育。我们研究青蛙的问题 非洲爪蟾，其丰富的卵是大的，容易通过显微注射和显微外科手术操作。的 胚胎足够大，可以产生生物化学和蛋白质组学分析的材料，也是理想的 成像整个胚胎形态发生和外植体的共聚焦成像。 在以前的资助期间，我们已经确定了有效的信号传导和信号转导活动， 有助于胚胎发育、神经诱导和神经板的前后图案化。在 与此同时，我们改进了X的基因组组装和注释。tropicalis和X. laevis，使系统级 approach.这些，以及其他青蛙的新组合，不仅为假设提供了必要的资源， 为社区推动研究，而且还提供了对基因组结构，重组和 进化 在下一个资助期，我们将重点放在控制细胞形状变化和组织水平运动的基因上。 胚胎，侧重于原肠胚和神经胚。基于Rho GTP交换因子 （GEF）通常选择性地用于介导特定的细胞行为，我们将继续分析 Plekhg家族的成员，其中七分之三在CRISPR后显示出特定的形态发生缺陷 介导的敲低，并将分析扩展到其他GEF或GT3激活蛋白（GAP）， 早期形态发生缺陷这些实验的结果将提供对正常的基本理解。 脊椎动物形态发生，以及对不同细胞行为控制方式的见解。

Project Summary/Abstract Our goal is to understand early vertebrate development at the molecular level. We study the problem in the frog Xenopus, whose abundant eggs are large and readily manipulated by microinjection and microsurgery. The embryos are large enough to produce material for biochemical and proteomic analysis, and are also ideal for imaging whole embryo morphogenesis, and confocal imaging of explants. During previous grant periods, we have identified potent signaling and signal transduction activities that contribute to embryonic development, neural induction and anterior-posterior patterning of the neural plate. In parallel we improved genome assemblies and annotation for X. tropicalis and X. laevis, enabling a systems level approach. These, and new assemblies for other frogs, not only provide the resources necessary for hypothesis driven research for the community, but also delivered new insights into genome structures, recombination and evolution. In the next grant period, we focus on the genes that control cell shape changes, and tissue level movements in the embryo, focusing on gastrulation and neurulation. Based on the premise that Rho GTP Exchange Factors (GEFs) are often deployed selectively to mediate specific cell behaviors, we will continue our analysis of members of the Plekhg family, among which three of seven show specific morphogenetic defects after CRISPR mediated knockdown, and extend the analysis to other GEFs or GTPase Activating Proteins (GAPs) that show early morphogenetic defects. The results of these experiments will provide basic understanding of normal vertebrate morphogenesis, and insights into the ways that different cellular behaviors are controlled.