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: 7991785
• 负责人: Jesús Torres-Vázquez
• 金额: $ 42.38万
• 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 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7991785
• 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. PUBLIC HEALTH RELEVANCE: The reproducible anatomical organization of the vaculature is essential for homeostasis and survival. PlexinD1 is an endothelial specific receptor that plays a key role in shaping the vascular pattern. In this proposal we address the molecular and cellular mechanism of vascular patterning by PlexinD1 signaling using zebrafish.

描述（由申请人提供）：早期胚胎血管形成具有良好的解剖学再现性，以确保体内平衡和存活。人们对脉管系统如何获得其典型的结构知之甚少。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）血管的发育。这些血管具有简单的模式，由少量内皮细胞组成，其行为很容易观察，并且可以使用遗传、化学和胚胎学工具进行研究。我们已经证明，内皮特异性 PlexinD1 (PlxnD1) 受体感知的旁分泌、排斥性信号蛋白 (Sema) 信号塑造了 Se 血管解剖结构。 Sema-PlxnD1 信号传导期间内皮细胞内部发生哪些分子事件？细胞内 PlxnD1 功能的机制基础和所涉及的分子参与者尚不清楚。 PlxnD1 具有类似于轴突 Plxns 中排斥活性所必需的胞质结构域以及 PlxnD1 特异性序列特征。这些基序是否是 PlxnD1 血管模式活性所必需的仍有待探索。因此，在具体目标 1 中，我们将询问这些结构域是否是 PlxnD1 介导的 Se 血管模式所必需的，并定义它们的细胞和生化作用。接下来，我们将表征 gipc1，它是第一个确定的 PlxnD1 信号传导候选调节器/效应器。 gipc1 在发育中的血管中特异性转录，它编码一种与 PlxnD1 胞质尾部物理关联的蛋白质，其活性对于 Se 血管模式至关重要。因此，在具体目标 2 中，我们将确定 GIPC1 对于 PlxnD1 介导的 Se 血管模式的分子和细胞作用。为此，我们将定义介导其与 PlxnD1 物理关联的 GIPC1 结构域，询问这种相互作用对于 PlxnD1 介导的 Se 血管模式是否是必要的，定义 GIPC1 是否促进或拮抗 PlxnD1 信号传导，并分析具有减少的动物的 Se 血管模式表型或 gipc1 水平升高。最后，在具体目标 3 中，我们重点关注毁灭之路 (rtp)，这是一种具有与 plxnD1 null 类似的 Se 血管缺陷的突变体。我们将从克隆 rtp 开始，探索它与 PlxnD1 信号传导的关系。然后，我们将确定 rtp 依赖性的内皮细胞行为。我们的研究有望为塑造健康和疾病中血管树的遗传程序和细胞行为提供重要见解。公共健康相关性：脉管系统的可重复解剖组织对于体内平衡和生存至关重要。 PlexinD1 是一种内皮特异性受体，在塑造血管模式中发挥关键作用。在本提案中，我们通过使用斑马鱼的 PlexinD1 信号传导来解决血管模式的分子和细胞机制。