Cell-type specific role of circadian-dependent transcription in fentanyl-induced synaptic and behavioral plasticity - Supplement

项目总结 睡眠和昼夜节律紊乱在阿尔茨海默病中非常普遍，比认知早几十年出现 拒绝。来自动物和人类的证据表明，这些干扰直接导致阿尔茨海默病 进一步加剧睡眠和昼夜节律失调的病理。研究这些问题的一个重要突破 Connections是最近开发的一种遗传多样化的小鼠面板，它包含了高危家族性 阿尔茨海默病突变(称为AD-BXD)概括了人类阿尔茨海默病的关键方面 病理生理学，包括衰老相关的神经变性、进行性认知缺陷和睡眠障碍。 利用这些小鼠，发现了一种新的遗传易感性标记，易受认知能力下降和睡眠障碍的影响 阿尔茨海默病：瞬时受体电位非选择性阳离子通道3型(TRPC3)。TRPC3有 已经被认为是改变正常认知老化和阿尔茨海默氏症发展的目标 疾病。我们发现，病毒介导的TRPC3基因敲除可减少淀粉样蛋白负荷并增强认知能力 在易感的AD-BXD小鼠中，为研究连接的机制链接提供了临床前基础 阿尔茨海默病患者的睡眠障碍和认知能力下降。然而，这些方法不能提供生物学 TRPC3通过其调节与疾病相关的病理改变。我们建议全面绘制空间地图 TRPC3的定位和表达以确定这种定位是否在关键脑区发生变化 与阿尔茨海默氏症的病理有关的睡眠和认知。我们开发了一种快速成像的新方法 全脑内阿尔茨海默病神经变性的多种细胞类型和标志物的三维(3D) 单元格分辨率的空间。我们的超高速高分辨率共聚焦带状扫描方法达到了 衍射限制分辨率(~200 NM)，并在不到24小时内收集3D渲染的全脑地图。这个 该附录的目标是利用TRPC3的发现作为阿尔茨海默氏症相关的新靶点 认知和睡眠的变化，并使用我们的创新工具回答基本问题：在小鼠身上，1)是 TRPC3位于与认知和睡眠有关的疾病相关的大脑区域？2)TRPC3与 阿尔茨海默病脑病理的生物学特征，包括淀粉样β蛋白和过度磷酸化 陶先生？我们将用人类死后阿尔茨海默病患者的大脑来问，3)大脑是TRPC3 阿尔茨海默病患者的表情会改变吗？我们预测TRPC3定位于下丘脑的亚区 与睡眠相关，以及与认知相关的海马区和皮质区域，这种分布将 易受睡眠剥夺的小鼠与有弹性的小鼠受到不同程度的影响。在人类大脑中，我们期望 在晚期阿尔茨海默病患者的大脑中，TRPC3的表达会更高，并表现出节律性的改变。 最终，使用我们新颖的基因、成像和计算工具套件，我们将回答长期存在的问题 关于睡眠变化和认知能力下降如何与阿尔茨海默病的病理联系的问题。

PROJECT SUMMARY Sleep and circadian disruption are highly prevalent in Alzheimer’s disease, emerging decades prior to cognitive decline. Evidence from animals and humans suggests these disruptions directly lead to Alzheimer’s disease pathology that further exacerbate sleep and circadian dysfunctions. An important breakthrough for studying these connections was the recent development of a genetically diverse mouse panel that incorporates high-risk familial Alzheimer’s disease mutations (termed AD-BXD) that recapitulate key aspects of human Alzheimer’s pathophysiology, including aging-related neurodegeneration, progressive cognitive deficits, and sleep disruption. Using these mice, identified a new marker of genetic vulnerability to cognitive decline and sleep disruption in Alzheimer’s disease: the transient-receptor potential nonselective cation channel type 3 (TRPC3). TRPC3 has already been implicated as a target for modifying the development of normal cognitive aging and Alzheimer’s disease. We found that viral-mediated knockdown of TRPC3 diminished amyloid load and enhanced cognition in susceptible AD-BXD mice, providing the preclinical basis for investigating the mechanistic links that connect sleep disruption and cognitive decline in Alzheimer’s disease. However, these methods do not offer the biology by which TRPC3 moderate disease-related pathology. We propose to comprehensively map the spatial location and expression of TRPC3 to identify whether this localization changes in key brain regions related to sleep and cognition due to Alzheimer’s pathology. We developed a new approach to rapidly image multiple cell-types and markers of Alzheimer’s neurodegeneration within a whole brain in three-dimensional (3D) space at single-cell resolution. Our ultra-fast high-resolution confocal ribbon-scanning approach reaches diffraction limited resolution (~200nm) and collects 3D rendered whole brain maps in less than 24-hours. The goals of the supplement are to take advantage of the discovery of TRPC3 as a new target for Alzheimer’s-related changes in cognition and sleep and use our innovative tools to answer fundamental questions: In mice, 1) Is TRPC3 located in disease-related brain areas linked to cognition and sleep?; and 2) Does TRPC3 interact with biological hallmarks of Alzheimer’s disease brain pathology, including amyloid beta and hyperphosphorylated Tau? We will use human postmortem brains from Alzheimer’s disease patients to ask, 3) Is brain TRPC3 expression altered in Alzheimer’s disease? We predict TRPC3 localizes to subregions of the hypothalamus associated with sleep and hippocampal and cortical regions associated with cognition, and this distribution will be differentially impacted by sleep deprivation in susceptible vs. resilient mice. In human brains, we expect that TRPC3 expression will be higher in advanced Alzheimer’s disease patient brains and display altered rhythmicity. Ultimately, using our novel suite of genetic, imaging, and computational tools, we will we will answer longstanding questions about how changes in sleep and cognitive decline are linked to Alzheimer’s disease pathology.