Super-Resolution Microscopy of Small Quantum Dots to Elucidate the Mechanisms of Alzheimer's Disease

小量子点的超分辨率显微镜阐明阿尔茨海默病的机制

• 批准号: 9274105
• 负责人: Hee Jung Chung
• 金额: $ 66.86万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-15 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9274105
• 关键词: AMPA Receptors; Acute; Affect; Alzheimer' s Disease; Alzheimer' s disease model; American; Amyloid beta-Protein; Antibodies; Behavior; Binding; Biochemical; Biological Assay; Brain; Brain Diseases; Caliber; Cells; Clinical; Collaborations; Color; Communication; Complex; Cultured Cells; Data; Deposition; Development; Dimensions; Disease; Electrophysiology (science); Engineering; Excitatory Synapse; Exhibits; Extracellular Domain; Fluorescent Dyes; Fostering; Generations; Glutamate Receptor; Glutamates; Goals; Grant; Hippocampus (Brain); Image; Imaging Techniques; Imaging technology; Immunoglobulin Fragments; Impaired cognition; Impairment; Individual; Intercellular Junctions; Label; Lead; Learning; Ligands; Long-Term Depression; Long-Term Potentiation; Manuscripts; Mediating; Membrane; Memory; Memory Loss; Methodology; Methods; Microscopy; Molecular; Mus; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Nerve Degeneration; Neurobiology; Neurologic; Neurons; Optical Methods; Optics; Pathogenesis; Pharmaceutical Preparations; Photons; Positioning Attribute; Probability; Proteins; Publishing; Quantum Dots; Rattus; Research; Research Personnel; Resolution; Sampling; Semiconductors; Senile Plaques; Series; Signal Transduction; Slice; Specificity; Streptavidin; Synapses; Synaptic Transmission; Synaptic plasticity; Techniques; Thick; Tissues; Transgenic Organisms; Wild Type Mouse; Work; abeta oligomer; aerobic respiration control protein; base; brain tissue; direct application; effective therapy; fluorescence imaging; fluorophore; high resolution imaging; improved; insight; light microscopy; molecular imaging; mouse model; nanoparticle; nanoscale; new technology; novel; novel therapeutic intervention; prevent; receptor; single molecule; trafficking; transgenic model of alzheimer disease; two-photon

PROJECT SUMMARY / ABSTRACT Alzheimer's disease (AD) afflicts more than 5 million Americans, yet no known drug is able to prevent or stop the disease. Before AD fully develops with insoluble amyloid-β plaque deposits and neurodegeneration, there is a progressive cognitive decline associated with the impairment of synaptic plasticity that underlies learning and memory. This abnormal synaptic plasticity is likely caused by soluble amyloid-β oligomers affecting the synaptic levels of AMPA and NMDA receptors, two glutamatergic receptors that mediate induction and expression of synaptic plasticity. However, the underlying detailed mechanisms are not known and are exceptionally challenging to study due to the complex behavior of these receptors and the small nanometer-scale dimensions of the synaptic domains in which they reside. The goal of this proposal is to understand the molecular details of abnormal synaptic plasticity present in early AD by developing small nanoparticle-based optical probes and new microscopy techniques to analyze the position and dynamics of AMPA and NMDA receptors in normal and AD brains. This goal will be accomplished through the individual and collective efforts of three principle investigators, Paul Selvin (microscopy), Andrew Smith (quantum dots) and Hee Jung Chung (neurobiology). They have previously worked as a team to publish two manuscripts on generating small quantum dots (sQD) (< 10 nm diameter) that can enter the neuronal synapse and accurately follow the receptor number and dynamic placement in dissociated cultured neurons. To achieve this goal, Aim 1 will optimize super-resolution imaging techniques for sQDs in dissociated hippocampal culture and thick hippocampal slices with intact circuitry, specifically focusing on 1- and 2-photon excitation with FIONA and PALM/STORM microscopy. This will allow < 20 nm resolution in all three dimensions. Aim 2 will develop a novel set of sQDs that are smaller, stable, and monovalent with minimal non-specific interaction with tissue. Aim 3 will apply sQDs and super-resolution optical methods to perform single-molecule imaging of glutamate receptors during synaptic plasticity in hippocampal culture and acute slices from wild-type and AD transgenic model mice. Because of our on-going successful collaboration, we are able to work with the AD model immediately, while new microscopy and quantum dots are being generated. This research will increase our understanding of the early pathogenesis of AD and therefore foster the development of new therapeutic strategies that could specifically inhibit the progression of cognitive decline of this disease.

项目摘要 /摘要 阿尔茨海默氏病（AD）折磨了超过500万美国人，但没有已知的药物能够预防或停止 疾病。在使用不溶性淀粉样蛋白斑块沉积和神经变性充分发展之前，有 与学习和研究基础的突触可塑性受损相关的渐进认知下降和 记忆。这种异常突触可塑性可能是由影响突触的固体淀粉样蛋白β低聚物引起的 AMPA和NMDA受体的水平，两个介导和表达的谷氨酸能受体 突触可塑性。但是，基本的详细机制尚不清楚，并且是异常的 由于这些受体的复杂行为和小纳米尺度的尺寸，研究的挑战 它们所在的突触域。 该提议的目的是了解早期存在异常突触可塑性的分子细节 通过开发基于纳米颗粒的小型光学问题和新的显微镜技术来分析AD AMPA和NMDA受体在正常和AD大脑中的位置和动力学。这个目标将实现 通过三个主要调查人员的个人和集体努力，保罗·塞尔文（显微镜），安德鲁 史密斯（量子点）和Hee Jung Chung（神经生物学）。他们以前曾担任团队发表 可以进入神经元的两个有关产生小量子点（SQD）（<10 nm直径）的手稿 突触并准确跟随受体数量和动态放置在解离的培养神经元中。 为了实现这一目标，AIM 1将优化分离的SQD的超分辨率成像技术 海马培养和厚的海马切片，具有完整的电路，专门针对1片和2光子 fiona和棕榈/风暴显微镜的兴奋。这将允许在所有三个维度上分辨率<20 nm。 AIM 2将开发一组新型的SQD，这些SQD较小，稳定且单价为最少的非特异性SQD 与组织相互作用。 AIM 3将应用SQD和超分辨率光学方法来执行单分子 在海马培养和野生型急性切片中突触可塑性期间谷氨酸受体的成像 和AD转基因模型小鼠。由于我们持续的成功合作，我们能够与 AD模型立即生成新的显微镜和量子点。这项研究会 提高我们对AD早期发病机理的理解，从而促进新的发展 可以专门抑制该疾病认知下降的进展的治疗策略。