Polarized Protein Trafficking and Angiogenesis

摘要 血管携带氧气和营养物质，对生物体的生存能力和持续的动态平衡至关重要。 血管生成，即从原有血管形成新的血管，是主要的发育过程。 调节血管网络密度的过程。在血管生成发育过程中，内皮细胞 创建一个称为管腔的中空腔，为血液到达远处的组织提供一个连续的管道。这个 支持内皮结构和信号的形态动力学变化的机制导致 血管管腔的形成，或小管的形成，尚不完全清楚。在这项提案中，我们将调查 一种叫做突触素样蛋白2(Syl 2)的蛋白质，我们认为它是导致管腔表面defi的原因。 在血管发育过程中，通过引导蛋白质运输到顶膜。我们的初步数据 提示Sytl2 defi不存在于顶膜上，并连接Rab GTP酶蛋白以传递囊泡。 货物，如足叶黄素。在目标1中，我们将描述sytl2a在血管腔形成中的作用。 利用活体成像和基于crispr的突变世代相结合的方法开发斑马fish胚胎。在……里面 目标2，我们将全面演示Sirt2与GTPase Rab35的结合作用，以实现 在体外管腔形成过程中，Podocalyin对顶端质膜的影响。在目标3中，我们将进一步 利用新斑马fish的生成研究了sytl2a如何与Rab35相互作用来传递podocalyin。 体内报道系和复合突变体。如何调节血管管腔的形成仍然是一个主要的问题 在fi领域，这一提议将为协调这一过程的关键机制提供新的见解。

SUMMARY Blood vessels carry oxygen and nutrients and are vital to organismic viability and continued homeostasis. Angiogenesis, or the formation of new blood vessels from pre-existing ones, is the predominant developmental process by which blood vessel network density is regulated. During angiogenic development, endothelial cells create a hollow cavity called a lumen, providing a continuous conduit for blood to reach distant tissues. The mechanisms underpinning the morphodynamic changes in endothelial architecture and signaling leading to vascular lumen formation, or tubulogenesis, are incompletely understood. In this proposal we will investigate a protein called synaptotagmin-like protein 2 (sytl2) that we believe is responsible for defining the luminal surface by directing protein transport to the apical membrane during blood vessel development. Our preliminary data suggests that sytl2 defines the apical membrane and tethers Rab GTPase proteins for delivery of vesicular cargo, such as podocalyxin. In aim 1, we will characterize the role of sytl2a during vascular lumen formation in developing zebrafish embryos using a combination of live-imaging and CRISPR-based mutant generation. In aim 2, we will comprehensively demonstrate that sylt2 works in combination with the GTPase Rab35 to deliver podocalyxin to the apical plasma membrane during lumenogenesis in vitro. In aim 3, we will further characterized how sytl2a interacts with Rab35 to deliver Podocalyxin using generation of new zebrafish reporter lines and compound mutants in vivo. How blood vessel lumen formation is regulated is still a major question in the field, this proposal will provide novel insight into critical mechanisms orchestrating this process.