Molecular principles of neuronal maturation and integration in the adult and aging brain

成人和衰老大脑中神经元成熟和整合的分子原理

• 批准号: 10159316
• 负责人: Paola Arlotta
• 金额: $ 53.35万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10159316
• 关键词: Acceleration; Address; Adult; Aging; Argentina; Argentine; Automobile Driving; Behavioral; Bioinformatics; Biological Assay; Birth; Brain; Brain region; Cell Maturation; Cells; Collaborations; Communities; Date of birth; Development; Developmental Delay Disorders; Disease; Environment; Epigenetic Process; Equipment; Exhibits; Exposure to; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Genetic Transcription; Global Change; Goals; Grant; Hippocampus (Brain); Human Resources; Individual; Institutes; Knowledge; Laboratories; Life; Mediating; Mediator of activation protein; Molecular; Molecular Profiling; Morphology; Mus; Neurons; Phase; Physical Exercise; Population; Postdoctoral Fellow; Process; Proteins; Research; Research Personnel; Role; Signal Transduction; Site; Space Perception; Stimulus; Techniques; Technology; Testing; Training; Work; aged; aging brain; aging hippocampus; dentate gyrus; differential expression; equipment training; experience; graduate student; granule cell; improved; in vivo; international center; knock-down; member; middle age; overexpression; prevent; tool; trait; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; young adult

The hippocampus is a region of the brain that continues to produce new dentate granule cells (GCs) throughout life. Development of adult-born GCs and their integration into preexisting circuits is modulated by environment and by electrical activity of the local circuits, and is altered in the aging brain. To understand how GC integration occurs and how it is modulated by activity and aging, it is crucial to dissect the precise molecular mechanisms underlying these processes. The technology required for addressing these fundamental problems is unavailable in Argentina, but routinely applied in the Arlotta lab at Harvard. Joining efforts to address this specific problem is a natural next step. The overarching goal of this proposal is to exploit the strategies for transcriptional profiling and bioinformatic analysis in brain development validated by the Arlotta lab and the Schinder lab's expertise in functional characterization of adult-born GCs to build an experimental pipeline for the discovery of new molecules controlling circuit plasticity in the adult and aging brain, that includes assays for testing the roles of individual proteins to an unprecedented level of molecular and functional detail. In Aim 1, we propose to generate a pipeline to reveal transcription factors, epigenetic regulators, or effector genes controlling the developmental transitions along GC maturation and integration. The proposed experimental pipeline is similar for all three Aims: (i) FACS-purify birth-dated adult-born GCs at different stages; (ii) transcriptionally profile each population using two complementary forms of RNA sequencing; (iii) bioinformatically identify transcription factors or epigenetic regulators that may control stage progression; and (iv) functionally test candidate molecules through in vivo knock-down or overexpression. Using this same approach, we will then investigate how stage-specific transcriptome dynamics are altered in GCs from the aging hippocampus, to identify and functionally test changes in regulatory molecules that may be responsible for their protracted development (Aim 2). Finally, we will seek to identify molecular mediators of activity-mediated acceleration in GC development in the adult and the aging brain. (Aim 3). This grant will further the aims of the Fogarty International Center in expanding the technical capacities of Dr Schinder's lab, including availability of equipment, training of Argentinian graduate students and postdoctoral fellows, and building collegial networks between members of the Leloir and Harvard research communities. This work will significantly expand the capacity of Leloir to apply state-of-the-art transcriptomic profiling (currently limited, throughout Argentina) to solve new problems related to brain function and disease, locally supported by a bioinformatician who is co-investigator in the project. The workflow proposed will therefore not only enable a new generation of molecular studies in the Schinder lab by implementing the most advanced molecular tools and technologies, but also transfer expertise and know-how through personnel training such that these technologies and approaches will become available for routine use at the Leloir Institute at large.

海马体是大脑中不断产生新的齿状颗粒细胞 (GC) 的区域 一生。成年 GC 的发育及其与先前存在的电路的整合受到以下因素的调节 环境和局部电路的电活动，并且在衰老的大脑中发生改变。要了解如何 GC 整合的发生以及它如何受活性和老化的调节，剖析精确的整合至关重要 这些过程背后的分子机制。解决这些问题所需的技术 阿根廷无法解决基本问题，但哈佛大学的阿洛塔实验室经常应用这一问题。加盟 下一步自然要努力解决这一具体问题。该提案的总体目标是利用 大脑发育中的转录谱和生物信息学分析策略已得到验证 Arlotta 实验室和 Schinder 实验室在成年 GC 功能表征方面的专业知识可构建 发现控制成人和衰老电路可塑性的新分子的实验流程 大脑，其中包括将单个蛋白质的作用测试到前所未有的分子水平的分析 和功能细节。在目标 1 中，我们建议建立一个管道来揭示转录因子、表观遗传学 调节器或效应基因控制 GC 成熟和整合的发育转变。 拟议的实验流程对于所有三个目标都是相似的：（i）FACS 纯化出生日期的成年出生 GC 不同阶段； (ii) 使用两种互补形式的 RNA 对每个群体进行转录分析 测序； (iii) 以生物信息学方式识别可能控制阶段的转录因子或表观遗传调节因子 进展； (iv) 通过体内敲低或过表达对候选分子进行功能测试。 使用相同的方法，我们将研究阶段特异性转录组动态如何在 来自老化海马体的 GC，用于识别和功能测试可能会影响的调节分子的变化 对其长期发展负责（目标 2）。最后，我们将寻求识别分子介质 活动介导的成人和衰老大脑中 GC 发育的加速。 （目标 3）。这笔赠款将 进一步实现福格蒂国际中心扩大 Schinder 博士实验室技术能力的目标， 包括设备的可用性、阿根廷研究生和博士后研究员的培训，以及 在勒卢瓦和哈佛大学研究社区的成员之间建立学院网络。这项工作将 显着扩展 Leloir 应用最先进的转录组分析的能力（目前有限， 整个阿根廷）解决与大脑功能和疾病相关的新问题，并得到当地的支持 生物信息学家，该项目的联合研究员。因此，所提出的工作流程不仅能够实现 Schinder 实验室采用最先进的分子工具进行新一代分子研究 和技术，还通过人员培训转让专业知识和技术，以便这些 技术和方法将可供整个勒卢瓦研究所日常使用。