3D Reconstruction and Analysis of Alzheimer's Patient Biopsy Samples to Map and Quantify Hallmarks of Pathogenesis and Vulnerability

项目概要/摘要 该项目将扩大三维电子显微镜的获取、重建、分析和传播， (3D EM）参考数据，揭示了保存在一个显着的遗产集合中的关键超微结构细节。 来自患有阿尔茨海默病（AD）的患者的活组织检查脑样品。这些样本最初是 由神经病理学家R.D.特里和N. Gonatas（在A.爱因斯坦在 1960年代），后来的样品作为脑脊液（CSF）药物输注研究的一部分，涉及S。米拉（在 1980年代的埃默里）。在20世纪80年代，Ellisman，Masliah，Terry和Mirra重新研究了它们，使用早期 3D EM方法，并发现表现出良好的超微结构保存，显示成对的螺旋 纤维（PHF）和淀粉样蛋白积累以及亚细胞器和细胞骨架的修饰 轴突和树突的突起。在这里，我们将利用高通量的最新进展， 自动化3D EM来大规模地检查这些珍贵的样本，重建100个大脑 有和没有PHF的细胞，通过更大的大脑体积跟踪轴突（并映射神经胶质和突触） 比以前可行。我们的目标是靶向与斑块和缠结相关的区域， 现有发现表明细胞和网络脆弱性并包含分子相互作用的位置 一些人怀疑这是AD发生和发展的基础。支持调查 索马/树突状变性的进展，我们将靶细胞操作定义为代表进展 在AD中观察到的神经元衰退;确定细胞质中PHF的体积分数作为一个实用的分期 测量并将其与主要亚细胞微结构中的定量变化的表征联系起来， 选民。同样，我们将分析斑块内和斑块附近轴突变性的进展作为数据 研究表明，轴突可能在其母细胞体和树突之前变得营养不良 堕落

PROJECT SUMMARY/ABSTRACT This project will expand the acquisition, reconstruction, analysis, and dissemination of 3D electron microscopic (3D EM) reference data, disclosing key ultrastructural details preserved within a remarkable collection of legacy biopsy brain samples from patients suffering from Alzheimer’s Disease (AD). These samples were originally collected, characterized and archived by neuropathologists R.D. Terry and N. Gonatas (at A. Einstein in the 1960’s), with later samples taken as part of a cerebrospinal fluid (CSF) drug infusion study involving S. Mirra (at Emory in the 1980’s). They were re-examined by Ellisman, Masliah, Terry, and Mirra in the 1980s, using early 3D EM methods, and were found to manifest excellent preservation of ultrastructure, showing paired helical filaments (PHF) and amyloid accumulations as well as modifications to subcellular organelles and cytoskeletons of the cell bodies, axonal and dendritic processes. Here, we will exploit recent advances in high throughput, automated 3D EM to massively scale the examination of these precious samples, reconstructing 100s of brain cells with and without PHF, tracking axons (and mapping glia and synapses) through much greater brain volumes than was feasible previously. Our goal is to target areas associated with both plaques and tangles, attending to locations where existing findings suggest cell and network vulnerability and contain molecular interactions suspected by some to underlie the initiation and progression of AD. Supporting investigations into the progression of soma/dendritic degeneration, we will target cells operationally defined to represent a progression of neuronal decline as seen in AD; determining the volume fraction of PHF in the cytoplasm as a practical staging measure and linking this to the characterization of quantitative changes in the microstructure of major subcellular constituents. Likewise, we will analyze the progression of axonal degeneration in and near plaques as data obtained suggests that axons may become dystrophic before their parent cell bodies and their dendrites degenerate.