Finding a molecular signature for dendrite regeneration

 描述(申请人提供)：树突可在中风、癫痫发作和创伤性脑损伤中受损。目前尚不清楚树突再生是否是从这些事件中恢复的正常部分。树枝晶再生可能是一个未知的过程，以提高回收率为目标。两个主要障碍阻碍了对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推移来研究树突再生；2.调节树突再生的机制完全未知。在这项提案中，我们建议突破这些障碍。在复杂组织损伤后，需要分子标记来研究树突再生。这项提议的第一个目标是识别树突再生的分子特征，可以用来精确定位树突再生发生在任何动物的任何组织中的时间和地点。为了定义树突再生特征，我们将从整个动物中分离出正在进行树突再生的单个神经元。我们将把这些细胞的转录本与未损伤的神经元和正在进行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类型的神经元中上调。在这次改进之后，我们将 测试商业上可用的抗体，以确定哪些标记将作为识别再生树突状细胞的特征集的一部分最有用。有了这个标记物，就有可能确定树突再生是由中风、癫痫发作还是创伤性脑损伤引发的。 为了确定树突再生在哪里是重要的，并了解我们可能如何操纵它来改变损伤结果，我们需要知道它是如何发挥作用的。第二个目标是确定树突再生的关键调控因素。在果蝇中无偏见的功能方法，以及基于转录组数据的筛选，将被用于识别控制树突再生的蛋白质。由于还没有鉴定出树突再生所需的基因，这一目标必然是探索性的。通过在一个模型系统中使用三种方法和现象级的遗传工具，我们将最大限度地增加我们识别树枝晶再生机制的第一块的机会。树突再生可能是受损神经系统恢复的主要因素。与轴突再生不同，树突再生没有分子参与者，关于树突再生的基本信息也很少。通过识别与树突再生相关和需要的第一个分子，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.