Gene Expression in Amphibian Development

项目摘要/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。我们在青蛙身上研究这个问题 非洲爪哇，其丰富的卵子很大，很容易通过显微注射和显微外科手术进行操作。这个 胚胎足够大，可以产生生化和蛋白质组分析的材料，也是理想的 全胚胎形态发生成像，外植体共聚焦成像。 在之前的赠款期间，我们已经确定了有效的信号和信号转导活动， 有助于胚胎发育、神经诱导和神经板的前后构型。在……里面 同时，我们改进了热带和莱维氏X.laevis的基因组组装和注释，使系统水平成为可能 接近。这些，以及其他青蛙的新组装，不仅为假说提供了必要的资源 推动社区研究，但也提供了对基因组结构、重组和 进化论。 在接下来的资助期间，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，主要集中在原肠形成和神经形成。基于Rho GTP交换因子的前提 (GEF)经常被选择性地部署来调节特定的细胞行为，我们将继续分析 Plekhg家族的成员，其中7个中的3个在CRISPR后显示出特定的形态发生缺陷 介导基因敲除，并将分析扩展到其他GEF或GTP酶激活蛋白(GAP)，表明 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 脊椎动物的形态发生，以及对不同细胞行为控制方式的洞察。

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.