Finding a molecular signature for dendrite regeneration

 描述（由申请人提供）：树突可能在中风、癫痫发作和创伤性脑损伤中受损。目前尚不清楚枝晶再生是否是从这些事件中恢复的正常部分。枝晶再生可能是一个未开发的过程，以提高采收率为目标。两个主要的障碍阻碍了树突再生在神经系统修复中的重要性的评估：1。目前只能通过跟踪单个神经元随时间的变化来研究树突再生，2。介导树突再生的机制是完全未知的。在本提案中，我们建议突破这些障碍。 研究复杂组织损伤后的树突再生需要分子标记。这项提议的第一个目的是确定树突再生的分子特征，可以用来确定它在任何动物的任何组织中何时何地发生。 为了定义树突再生特征，我们将从整个动物中分离经历树突再生的个体神经元。我们将这些细胞的转录组与未损伤的神经元和经历轴突再生的神经元进行比较。我们将选择一组在树突再生中高度上调的10 - 15个基因，并确定它们在树突再生过程中是否也在不同的神经元类型中上调。经过这一改进，我们将 测试商业上可获得的抗体，以确定哪些标记物将作为识别再生树突的细胞的特征集的一部分最有用。有了这个标志物集，就有可能确定树突再生是否是由中风、癫痫发作或创伤性脑损伤引起的。 为了确定树突再生的重要性，并了解我们如何操纵它来改变损伤结果，我们需要知道它是如何工作的。第二个目标是集中在确定树突再生的关键调节。在果蝇中无偏见的功能方法，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，因此这一目标必然是探索性的。通过在模型系统中使用三种方法与现象遗传工具，我们将最大限度地提高我们的机会，确定第一件树突再生机制。 树突再生可能是受损神经系统恢复的主要参与者。与轴突再生不同，树突再生没有分子参与者，而且已知的基本信息很少。通过识别与树突再生相关并需要的第一个分子，该项目将为机制研究开辟一个新的领域。

 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.