Structure and function of mammalian CPEB2 aggregates in normal and AD brain

项目摘要 阿尔茨海默病的临床前和临床试验的总体失败表明，改善的基础 对记忆的理解以及记忆的分子成分在阿尔茨海默病中如何改变 这一过程对于开发有效的治疗方法是必要的。该项目的总体目标是确定 哺乳动物持久记忆的生化底物。目前的提案重点是一个家庭 RNA结合，细胞质多聚腺苷酸化元件结合蛋白(CPEB)，稳定记忆 无脊椎动物和老鼠。值得注意的是，CPEB家族蛋白形成非致病淀粉样变性 CPEB的聚合和聚合是稳定内存所必需的。因为淀粉样蛋白通常与 疾病状态下，问题仍然是结构相似的β42或Tau蛋白如何产生相反的影响 在记忆中。因此，为了更好地理解淀粉样蛋白之间的关系 记忆和淀粉样蛋白破坏记忆，我们将使用各种技术来解决结构和 CPEB家族成员CPEB2和CPEB3在人和小鼠中的功能。在目标1中，我们将使用冷冻- 用电子显微镜研究新鲜人额颞叶组织CPEB聚集体的结构 收集自25-50岁接受组织切除手术的人类受试者，标准为 疾病。否则这些纸巾就会被丢弃。在目标2中，小鼠缺乏形成能力 CPEB2和CPEB3的聚集体将在一次试行的抑制回避任务中进行训练和测试，以进行分析 他们形成、保持和回忆记忆的能力。在目标2中，我们还将调查CPEB2的后果 CPEB3聚集在编码突触蛋白的mRNA的翻译中。这一结果将是第一个 提供哺乳动物中与记忆相关的功能性淀粉样蛋白的直接结构分析，结构 人脑中功能性淀粉样蛋白和毒性淀粉样蛋白之间的区别，并精确地将CPEB2和 CPEB3的聚集和活性对动物形成或稳定记忆的能力。这一知识将提供 为未来研究有毒的淀粉样蛋白Aβ42或Tau如何特别干扰记忆奠定了基础。

Project Summary The aggregate failure of pre-clinical and clinical trials in AD has demonstrated that an improved fundamental understanding of memory and how the molecular components of memory are altered in the AD disease process is necessary to develop effective treatment. The broad objective of the project is to identify the biochemical substrates of long-lasting memories in mammals. The current proposal focuses on a family of RNA-binding, cytoplasmic polyadenylation element binding protein (CPEB), that stabilizes memory in invertebrates and mice. Remarkably, CPEB family protein forms non-disease-causing amyloidogenic aggregates and aggregation of CPEB is necessary to stabilize memory. As amyloids are typically linked to disease states, the question remains how similarly structured Aβ42 or Tau proteins can have opposing effects on memory. Therefore, to develop a better understanding of the relationship between amyloids that support memory and amyloids that disrupt memory, we will use a variety of techniques to solve the structure and function of the CPEB family members, CPEB2 and CPEB3, in human and mice. In Aim 1, we will use cryo- electron microscopy to solve the structure of CPEB aggregates from fresh human frontotemporal lobe tissue collected from 25-50-year-old human subjects undergoing tissue removal under the standard of care for their disease. These tissues would have been otherwise discarded. In Aim 2, mice lacking the ability to form aggregates of CPEB2 and CPEB3 will be trained and tested in a one-trial inhibitory avoidance task to assay their ability to form, maintain, and recall memory. In Aim 2 we will also investigate the consequence of CPEB2 and CPEB3 aggregation in translation of mRNA encoding synaptic proteins. The results would be the first to provide direct structural analysis of a functional amyloid linked to memory in mammals, the structural distinctions, if any, between functional and toxic amyloid in the human brain, and precisely link CPEB2 and CPEB3 aggregation and activity to animals’ ability to form or stabilize memory. This knowledge would provide the foundation to investigate in the future how toxic amyloids of Aβ42 or Tau specifically perturb memory.