Molecular and cellular mechanisms of vascular patterning by PlexinD1 signaling

PlexinD1 信号传导血管模式的分子和细胞机制

• 批准号: 8387036
• 负责人: Jesús Torres-Vázquez
• 金额: $ 39.94万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8387036
• 关键词: Address; Alleles; Anatomy; Animals; Architecture; Behavior; Biochemical; Blood Vessels; Cell Surface Receptors; Cloning; Cues; Defect; Development; Disease; Embryo; Endothelial Cells; Endothelium; Ensure; Event; Genes; Genetic; Genetic Programming; Genetic Screening; Growth; Health; Homeostasis; Mediating; Modeling; Molecular; Morphogenesis; Pathway interactions; Pattern; Phenotype; Play; Positioning Attribute; Proteins; Reproducibility; Role; Semaphorins; Shapes; Signal Transduction; Tail; Testing; Time; Tissues; Trees; Zebrafish; base; cell behavior; chemical genetics; in vivo; insight; member; mutant; novel; paracrine; receptor; tool; zebrafish development

DESCRIPTION (provided by applicant): Early embryonic blood vessels form with great anatomical reproducibility to ensure homeostasis and survival. How the vasculature acquires its stereotypical architecture is poorly understood. To uncover the genetic and cellular basis of vascular patterning we study the development of the zebrafish Segmental (Se) vessels. These vessels have a simple pattern, are made of few endothelial cells whose behaviors are easily observed and can be studied with genetic, chemical and embryological tools. We have shown that paracrine, repellent Semaphorin (Sema) cues sensed by the endothelial-specific PlexinD1 (PlxnD1) receptor shape the Se vessel anatomy. What molecular events occur inside the endothelial cell during Sema-PlxnD1 signaling? The mechanistic basis of intracellular PlxnD1 function and the molecular players involved are unknown. PlxnD1 has cytosolic domains similar to those essential in axonal Plxns for repulsive activity and also PlxnD1- specific sequence features. Whether these motifs are required for PlxnD1's vascular patterning activity remains unexplored. Thus, in Specific Aim 1 we will ask if these domains are required for PlxnD1-mediated Se vessel patterning and define their cellular and biochemical roles. Next, we will characterize gipc1, the first identified candidate modulator/effector of PlxnD1 signaling. gipc1 is specifically transcribed in the developing vessels, it encodes a protein that physically associates with the PlxnD1 cytosolic tail and its activity is essential for Se vessel patterning. Thus, in Specific Aim 2 we will determine the molecular and cellular roles of GIPC1 for PlxnD1-mediated Se vessel patterning. To do this we will define the GIPC1 domains that mediate its physical association with PlxnD1, ask if this interaction is necessary for PlxnD1-mediated Se vessel patterning, define if GIPC1 promotes or antagonizes PlxnD1 signaling and analyze the Se vessel patterning phenotypes of animals with reduced or increased levels of gipc1. Finally, in Specific Aim 3 we focus on road to perdition (rtp), a mutant with Se vessel defects similar to those of plxnD1 nulls. We will start by cloning rtp to explore its relationship to PlxnD1 signaling. Then, we will determine the endothelial cell behaviors that are rtp-dependent. Our studies promise to offer key insights into the genetic programs and cellular behaviors that shape the vascular tree in both health and disease.

描述（由申请人提供）：早期的胚胎血管形成，具有出色的解剖学可重复性，以确保体内平衡和生存。脉管系统如何获得其刻板印象的结构知之甚少。为了揭示血管模式的遗传和细胞基础，我们研究了斑马鱼节段（SE）血管的发展。这些容器具有简单的模式，由几个内皮细胞制成，其行为很容易观察到，并且可以使用遗传，化学和胚胎学工具进行研究。我们已经表明，旁分泌，驱虫词（SEMA）通过内皮特异性的Plexind1（PLXND1）受体所感受到SE血管解剖结构。在SEMA-PLXND1信号传导期间，内皮细胞内发生了哪些分子事件？细胞内PLXND1功能和所涉及的分子参与者的机械基础尚不清楚。 PLXND1具有与轴突PLXN中必不可少的胞质结构域的排斥活性和PLXND1-特定序列特征。 PLXND1的血管图案活性是否需要这些基序仍未得到探索。因此，在特定的目标1中，我们将询问PLXND1介导的SE容器的模式是否需要这些结构域，并定义其细胞和生化作用。接下来，我们将表征GIPC1，这是PLXND1信号传导的第一个确定的候选调制器/效应子。 GIPC1专门在发育中的血管中转录，它编码一种与PLXND1胞质尾巴物理相关的蛋白质，其活性对于SE血管构图至关重要。因此，在特定的目标2中，我们将确定PLXND1介导的SE血管模式的GIPC1的分子和细胞作用。为此，我们将定义介导其与PLXND1的物理关联的GIPC1结构域，询问这种相互作用是否对于PLXND1介导的SE容器模式性是必需的，定义GIPC1是否促进或拮抗PLXND1信号传导并分析SE SEEL模式性表型，具有降低GIPC1的动物或增加GIPC1的动物。最后，在特定的目标3中，我们专注于通往灭生道路（RTP），这是一种具有SE血管缺陷的突变体，类似于PLXND1 nulls。我们将从克隆RTP开始探索其与PLXND1信号传导的关系。然后，我们将确定依赖RTP的内皮细胞行为。我们的研究承诺将对塑造健康和疾病中血管树的遗传程序和细胞行为提供关键见解。