Cellular Anatomy of Tau Seeding

Tau 播种的细胞解剖学

• 批准号: 10575918
• 负责人: Sarah H Shahmoradian
• 金额: $ 24.6万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10575918
• 关键词: 3-Dimensional; Alzheimer' s Disease; Antibodies; Axon; Biosensor; Brain; Bypass; Cells; Cytoplasm; Data; Dendrites; Diamond; Electron Microscopy; Electrons; Event; Faculty; Human; Image; Immunofluorescence Immunologic; Impaired cognition; In Situ; Label; Light; Methods; Modeling; Molecular; Molecular Conformation; Molecular Machines; Monitor; Morphologic artifacts; Neurodegenerative Disorders; Neurons; Organelles; Pathologic; Pathology; Pattern; Presynaptic Terminals; Process; Proteins; Recombinants; Resolution; Resources; Role; Subcellular Anatomy; System; Techniques; Testing; Transgenic Mice; Visualization; human stem cells; imaging approach; induced pluripotent stem cell; light microscopy; live cell imaging; member; microscopic imaging; nanoscale; neuronal cell body; novel; protein aggregation; spatiotemporal; tau Proteins; tau conformation; tool; uptake

Project Summary/Abstract: The propagation of tau pathology along synaptically connected brain circuits is considered a major driver of cognitive decline in Alzheimer’s Disease (AD). However, the mechanisms underlying proteopathic tau seed entry and amplification within human neurons remain unclear. Given the putative role of brain circuits in tau propagation, axons/axon terminals or dendrites may be involved in tau seed uptake. However, this has never been determined in neuronal models, and typical methods to study these events, such as light microscopy and immunofluorescence alone, have critical resolution limits. Thus, we cannot yet hypothesize where in a neuron that the tau seeds access the cytoplasm and replicate. To answer these questions, we will apply high-resolution correlative light and electron microscopic imaging (3D-CLEM), combined with advanced cell-based fluorescent biosensors and novel conformational antibodies with live cell imaging in human neurons derived from inducible pluripotent stem cells (iPSCs) available here in our center. Our strategy should enable determining where tau seed uptake occurs, and the localization and sub-cellular components and/or molecular machinery involved in tau seed amplification, while bypassing protein aggregate extraction techniques that introduce experimental artifact. By exploring these mechanisms using high-resolution correlative imaging within human neurons, we will address how proteopathic seeds access the cytoplasm and replicate, which may have broader implications for other related neurodegenerative diseases.

项目概要/摘要： tau病理学沿着突触连接的脑回路的传播被认为是神经系统疾病的一个重要特征。 阿尔茨海默病（AD）认知能力下降的主要驱动因素。然而， 人类神经元内的蛋白病性tau种子进入和扩增仍不清楚。鉴于假定的 脑回路在tau传播中的作用，轴突/轴突终末或树突可能参与tau种子 摄取。然而，这从未在神经元模型中确定，并且典型的研究方法 这些事件（例如光显微镜和单独的免疫荧光）具有关键的分辨率极限。 因此，我们还不能假设在神经元中tau种子进入细胞质的位置， 复制为了回答这些问题，我们将应用高分辨率的相关光和电子 显微成像（3D-CLEM），结合先进的基于细胞的荧光生物传感器和新的 构象抗体与来自诱导性多能神经元的人神经元中的活细胞成像 我们中心提供干细胞（iPSC）。我们的策略应该能够确定tau蛋白的种子 发生摄取，并且涉及定位和亚细胞组分和/或分子机制 在tau种子扩增中，虽然绕过了引入 实验品通过使用高分辨率相关成像来探索这些机制， 人类神经元，我们将解决蛋白质种子如何进入细胞质和复制， 可能对其他相关的神经退行性疾病有更广泛的影响。