Finding a molecular signature for dendrite regeneration

寻找树突再生的分子特征

• 批准号: 8867657
• 负责人: Melissa Rolls
• 金额: $ 22.16万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8867657
• 关键词: Animal Model; Animals; Antibodies; Apoptosis; Biological Assay; Biological Models; Boxing; Cells; Complement; Complex; Data; Dendrites; Drosophila genus; Event; Excision; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Individual; Injury; Investigation; Label; Light; MAP Kinase Modules; MAPK8 gene; Mediating; Meta-Analysis; Methods; Mining; Molecular; Molecular Profiling; Morphology; Natural regeneration; Nervous System Trauma; Nervous system structure; Neurons; Outcome; Pathway interactions; Peripheral Nervous System; Phosphotransferases; Positioning Attribute; Preparation; Process; Proteins; RNA Interference; RNA Sequences; RNA library; Recovery; Role; Seizures; Signal Transduction; Stroke; Testing; Time; Tissues; Traumatic Brain Injury; Vertebrates; Visual; Work; axon injury; axon regeneration; base; cell type; dosage; improved; in vivo; injured; jun Oncogene; laser capture microdissection; molecular marker; new growth; prevent; public health relevance; repaired; research study; response; response to injury; tool; transcription factor; transcriptome sequencing

 DESCRIPTION (provided by applicant): Dendrites can be damaged in stroke, seizure and traumatic brain injury. It is not known whether dendrite regeneration is a normal part of recovery from any of these events. Dendrite regeneration may be an unexplored process to target for improving recovery. Two major impediments prevent assessment of the importance of dendrite regeneration in nervous system repair: 1. Dendrite regeneration can currently be studied only by tracking individual neurons over time, 2. The machinery that mediates dendrite regeneration is completely unknown. In this proposal we propose to break through these impediments. Molecular markers are required to study dendrite regeneration in complex tissues after they are damaged. The first aim of this proposal is to identify a molecular signature of dendrite regeneration that can be used to pinpoint when and where it occurs in any tissue in any animal. To define a dendrite regeneration signature we will isolate individual neurons undergoing dendrite regeneration from whole animals. We will compare transcriptomes of these cells to uninjured neurons and neurons undergoing axon regeneration. We will select a set of 10-15 genes that are highly upregulated in dendrite regeneration and determine whether they are also upregulated in a different neuron type during dendrite regeneration. After this refinement, we will test commercially available antibodies to determine which markers will be most useful as part of a signature set to identify cells regenerating dendrites. With this marker set, it will be possibleto determine whether dendrite regeneration is initiated by stroke, seizure or traumatic brain injury. To determine where dendrite regeneration is important, and to understand how we might manipulate it to change injury outcome, we need to know how it works. The second aim is focused on identifying key regulators of dendrite regeneration. Unbiased functional approaches in Drosophila, as well as a screen based on transcriptome data, will be used to identify proteins that control dendrite regeneration. As no genes required for dendrite regeneration have yet been identified this aim is necessarily exploratory. By using three approaches in a model system with phenomenal genetic tools we will maximize our chances of identifying the first pieces of the dendrite regeneration machinery. Dendrite regeneration is potentially a major player in the recovery of the damaged nervous system. Unlike axon regeneration, no molecular players and very little basic information about dendrite regeneration is known. By identifying the first molecules that are associated with and required for dendrite regeneration, this project will open a new field for mechanistic studies.

 描述（由适用提供）：树突在中风，癫痫发作和创伤性脑损伤中可能受损。尚不清楚树突再生是否是从任何这些事件中恢复的正常部分。树突再生可能是提高恢复目标的意外过程。两种主要障碍阻止了树突再生在神经系统修复中的重要性：1。目前只能通过跟踪单个神经元随着时间的推移，2。介导的树突再生的机制完全未知，目前只能通过跟踪单个神经元进行研究。在此提案中，我们建议打破这些障碍。分子标记需要在损坏后在复杂时机中研究树突再生。该提案的第一个目的是确定树突再生的分子特征，该分子特征可用于查明任何动物中任何组织中发生的何时何地发生。为了定义树突再生签名，我们将分离从整个动物的树突再生的单个神经元。我们将比较这些细胞的转录组与正在进行轴突再生的未受损神经元和神经元。我们将选择一组10-15个基因，这些基因在树突再生中高度更新，并确定它们在树突再生过程中是否也以不同的神经元类型进行更新。精炼之后，我们将 测试可商购的抗体，以确定哪些标记物作为签名集的一部分最有用，以识别再生树突的细胞。使用此标记集，可以确定是否通过中风，癫痫发作或创伤性脑损伤引发树突再生。 要确定树突再生在哪里很重要，并了解我们如何操纵它以改变伤害结果，我们需要知道其工作原理。第二个目的是确定树突再生的关键调节剂。果蝇中的无偏功能方法以及基于转录组数据的屏幕将用于识别控制树突再生的蛋白质。由于尚未确定树突再生所需的基因，因此这种目标一定是探索性的。通过在模型系统中使用具有惊人遗传工具的三种方法，我们将最大程度地识别识别树突再生机制的第一部分的机会。树突再生可能是恢复受损神经系统的主要参与者。与轴突再生不同，没有分子参与者和关于树突再生的基本信息很少。通过识别与树突再生相关和所需的第一个分子，该项目将为机械研究开放一个新的领域。