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.

项目摘要/摘要： tau 病理学沿着突触连接的大脑回路的传播被认为是 然而，阿尔茨海默病（AD）认知能力下降的主要驱动因素。 鉴于假定，tau 蛋白种子在人类神经元内的进入和扩增仍不清楚。 脑回路在 tau 传播中的作用，轴突/轴突末端或树突可能参与 tau 种子 然而，这从未在神经模型和典型的研究方法中得到确定。 这些事件，例如单独的光学显微镜和免疫荧光，具有关键的分辨率限制。 因此，我们还无法确定 tau 种子在神经元中的哪个位置进入细胞质并 为了回答这些问题，我们将应用高分辨率相关光和电子。 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颖的 在源自诱导多能的人类神经元中具有活细胞成像的构象抗体 我们中心提供的干细胞 (iPSC) 应该能够确定 tau 种子的位置。 发生摄取，并且涉及定位和亚细胞成分和/或分子机制 在 tau 种子扩增中，同时绕过引入了蛋白质聚集体提取技术 通过使用高分辨率相关成像来探索这些机制。 人类神经元，我们将解决蛋白病种子如何进入细胞质并复制，这 可能对其他相关的神经退行性疾病具有更广泛的影响。