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

项目摘要 睡眠和昼夜节律紊乱在阿尔茨海默病中非常普遍，比认知疾病早几十年出现 下降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功能障碍的病理学。研究这些的一个重要突破是， 连接是最近开发的一种遗传多样性小鼠面板， 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中断。 使用这些小鼠，确定了一种新的遗传易感性标记，以认知能力下降和睡眠中断， 阿尔茨海默病：瞬时受体电位非选择性阳离子通道3型（TRPC 3）。TRPC 3具有 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靶点 疾病我们发现，病毒介导的TRPC 3敲低减少了淀粉样蛋白负荷，增强了认知能力。 在易感的AD-BXD小鼠中，为研究连接 阿尔茨海默病患者睡眠中断和认知能力下降然而，这些方法不提供生物学 TRPC 3通过其调节疾病相关的病理。我们建议全面绘制 TRPC 3的定位和表达，以确定这种定位是否在关键脑区发生变化 与睡眠和认知有关的疾病。我们开发了一种新的方法来快速成像 在三维（3D）的全脑内阿尔茨海默氏神经变性的多种细胞类型和标志物 以单细胞分辨率间隔。我们的超快速高分辨率共焦带扫描方法达到 衍射极限分辨率（~ 200 nm），并在不到24小时内收集3D渲染的全脑地图。的 该补充剂的目标是利用TRPC 3的发现作为阿尔茨海默病相关疾病的新靶点。 认知和睡眠的变化，并使用我们的创新工具来回答基本问题： TRPC 3位于与认知和睡眠相关的疾病相关脑区？和2）TRPC 3是否与 阿尔茨海默病脑病理学的生物学标志，包括淀粉样蛋白β和过度磷酸化 陶？我们将使用阿尔茨海默病患者的人类死后大脑来问，3）大脑TRPC 3 在阿尔茨海默病中的表达改变我们预测TRPC 3定位于下丘脑的亚区域， 与睡眠相关，与认知相关的海马和皮层区域，这种分布将 睡眠剥夺对易感小鼠和适应性小鼠的影响不同。在人类大脑中， TRPC 3在晚期阿尔茨海默病患者大脑中的表达将更高，并显示出改变的节律性。 最终，利用我们新颖的遗传、成像和计算工具，我们将回答长期存在的问题。 关于睡眠和认知能力下降的变化如何与阿尔茨海默病病理学有关的问题。

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.