Netrin signaling during neuronal development

神经元发育过程中的 Netrin 信号传导

基本信息

批准号：
7804466
负责人：
ELKE STEIN
金额：
$ 33.74万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7804466
关键词：
Axon Binding Biochemical Biochemical Genetics Biological Assay Cells Complex Cues Cytoskeleton Destinations Development Embryo Event Family Foundations Genetic Goals Growth Cones Homodimerization Homologous Gene In Vitro Injury Invertebrates Lead Ligands Mammalian Cell Maps Mediating Messenger RNA Methods Molecular Nerve Regeneration Nervous system structure Neurites Neurons Orphan Proteins Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Route Signal Transduction Spinal Spinal Cord Staging Testing Therapeutic Intervention Translating Vertebrates Work Xenopus Yeasts axonal guidance cell motility developmental disease extracellular gain of function human NTN1 protein improved in vivo insight loss of function mRNA Expression member netrin receptor netrin-1 neurodevelopment neuron development novel programs receptor receptor function reconstitution research study response

项目摘要

DESCRIPTION (provided by applicant): In the developing nervous system, nerve cells and axons respond to a variety of attractive and repulsive guidance cues en route to their final destination. On their journey they follow a complicated trajectory using in some circumstances multiple cues, in others switching their responsiveness to a particular bifunctional cue, acting over short and long range. An essential step in obtaining a better understanding of the molecular mechanisms that underlie these events is to identify receptor complexes that couple guidance cues to directed guidance. Uncovering the molecular mechanisms that translate an extracellular cue into a change in the growth cone cytoskeleton is likely to lead to a better understanding of certain developmental disorders, and also to contribute towards development of therapeutic interventions to improve nerve regeneration following injury. The central aim of this proposal is to characterize receptor complexes that mediate responses of growth cones to the bifunctional guidance cue Netrin-1 in the developing nervous system.Netrin-1 attracts several classes of axons, while repelling others. Netrin receptors of the DCC family are implicated in mediating both attraction and repulsion; in contrast members of the UNC-5 family are implicated in signaling repulsion alone or in combination with DCC. How UNC5 signals repulsion independent of DCC is not known. Here we hypothezise that UNC5 signals two types of DCC-independent repulsion, through homomultimerization of UNC5A and through heteromultimerization of UNC5B-D with the orphan receptor PUNC. We are determined to elucidate how Netrin-1 mediates axonal repulsion. We will (I) characterize DCC- independent repulsion in vertebrates spinal neurites in vitro, (II) define the receptor complexes mediating DCC independent repulsion by using a combination of biochemical and genetic approaches and (III) evaluate the function of these receptors in Netrin-1 signaling in vitro and in vivo by utilizing directional and outgrowth responses of primary neuronal cultures in combination with loss- and gain-of-function approaches. This work will provide the foundation for understanding how neurons translate a bidirectional cue, such as Netrin-1, into directed guidance. In addition, it should be of more general importance in providing insights into the underlying receptor mechanism used by bifunctional cues to guide axons during neurodevelopment.

描述（由申请人提供）：在发育中的神经系统中，神经细胞和轴突在通往其最终目的地的途中对各种有吸引力和排斥的指导提示做出反应。在旅途中，他们遵循复杂的轨迹，在某些情况下使用多个提示，在其他情况下，他们将响应能力切换到特定的双功能提示，并在短距离和远距离上起作用。更好地了解这些事件基础的分子机制的重要步骤是确定将指导提示引导指导引导的受体复合物。揭示将细胞外提示转化为生长锥细胞骨架变化的分子机制可能会使人们更好地了解某些发育障碍，并有助于发展治疗干预措施以改善受伤后的神经再生。该建议的核心目的是表征受体复合物，以介导生长锥对发育中的神经系统中的双功能指导提示Netrin-1的反应。Netrin-1吸引了几类轴突，同时驱除了其他轴突。 DCC家族的Netrin受体与介导吸引和排斥有关。相比之下，UNC-5家族的成员与单独的信号拒绝或与DCC结合在一起。 UNC5信号如何独立于DCC信号。在这里，我们假设UNC5通过UNC5A的同源化和通过与孤儿受体PUNC的UNC5B-D异拟二聚化来指示两种类型的DCC依赖性排斥。我们决心阐明Netrin-1如何介导轴突排斥。我们（i）将在体外表征脊椎动物的脊髓神经突中的DCC-独立排斥，（ii）通过使用生物化学和遗传方法的组合来定义受体复合物，从而介导了DCC独立排斥，并通过在netrin-1信号中的构图中评估这些受体在体外和概要中的功能，并评估这些受体的功能，并构成了概述，并构成了概述，并将远期构成及列表结合构成，远期构成概述，远期概述概述，远期概述概述的概述，远期远程构成概述。和功能收益方法。这项工作将为理解神经元如何将双向提示（例如Netrin-1）转化为指导指导的基础。此外，在提供双功能提示在神经发育过程中使用的基本受体机制的见解应该更为重要。