Array Tomographic Single-Synapse Analysis of Normal and Disordered Cortex

描述（由研究者提供）：拟议的研究探讨了一种新的高分辨率组织分子成像方法，称为阵列断层扫描（AT）的有用性，以了解，诊断，预防和治疗阿尔茨海默病（AD）。 作为这项工作的一部分，一组新的阵列断层扫描分子标记和图像分析方法，称为阵列断层扫描单突触分析（AT/SSA），将被开发用于量化和分类小鼠和人类大脑和海马皮质组织中的单个突触的目的。 皮质突触的丧失被广泛认为是AD和许多其他神经退行性疾病中认知功能障碍的近端原因，但是由于可用方法的技术限制，关于这种皮质突触缺陷的定量信息有限。 此外，皮质突触现在被认为是高度多样化的，而且相关的定量信息也很少。 这些缺点对人类心理健康是重要的，因为有强有力的证据表明，不同的神经化学定义的突触类型在AD中表现出不同的病理学，因此代表了不同的治疗靶点发现机会。 AT/SAA有望提供有关皮质突触群体及其多样性的新的定量信息，这些信息可能填补我们对AD理解的关键空白，并导致新的诊断和治疗策略的发展。 AT/SSA是基于阵列断层免疫荧光成像的能力，以解决和量化的皮质神经元的三维体积中的单个突触点，并使用高阶抗体复用来区分各种分子定义的突触类型。 本项目将继续努力使阵列层析成像方法更快，更容易，更可靠和更丰富的信息，并开发高通量的阵列层析分类工具，用于区分和量化突触类型。 为了建立量化正常皮质突触群体的基线，AT/SSA方法将首先应用于来自野生型小鼠和来自未受AD影响的人的尸检和活组织检查的组织标本。 然后将相同的方法应用于转基因疾病模型小鼠和神经学特征的AD患者的标本。 所获得的信息有望为人类患者AD疾病进展的细胞和分子基础提供新的见解，并确定疾病模型小鼠可用于帮助开发和测试新药、疫苗或其他旨在预防和治愈AD的治疗方法的新方法。 公共卫生相关性：这项研究探讨了一种新的高分辨率分子成像技术，称为阵列断层扫描（AT），以了解，诊断，预防和治疗阿尔茨海默病（AD）的潜在用途。 将开发一套新的基于AT的方法，并用于比较大脑正常和AD受影响的人和正常小鼠和经遗传修饰以模拟人类AD状况的小鼠的皮质突触。 所获得的信息有望为人类患者AD疾病进展的细胞和分子基础提供新的见解，并确定疾病模型小鼠可用于帮助开发和测试新药、疫苗或其他旨在预防和治愈AD的治疗方法的新方法。

DESCRIPTION (provided by investigator): The proposed research explores the usefulness of a new high-resolution tissue molecular imaging method called Array Tomography (AT) to the understanding, diagnosis, prevention, and cure of Alzheimers Disease (AD). As part of this effort, a new set of array tomographic molecular labeling and image analysis methods, called Array Tomographic Single-Synapse Analysis (AT/SSA), will be developed for the purpose of quantifying and classifying individual synapses in tissues of mouse and human cerebral and hippocampal cortex. Loss of cortical synapses is widely believed to be the proximal cause of cognitive dysfunction in AD and many other neurodegenerative disorders, but quantitative information about such cortical synapse deficiencies has been limited, due to the technical limitations of available methodology. Moreover, cortical synapses are now recognized as being highly diverse, and again, relevant quantitative information is scarce. These shortcomings are consequential to human mental health because of strong evidence that different neurochemically defined synapse types exhibit differential pathologies in AD and therefore represent distinct therapeutic target discovery opportunities. AT/SAA is expected to provide new quantitative information about cortical synapse populations and their diversity that could fill critical gaps in our understanding of AD and lead to the development of novel diagnostic and therapeutic strategies. AT/SSA is based on the ability of array tomographic immunofluorescence imaging to resolve and quantify individual synaptic puncta in three-dimensional volumes of cortical neuropil, and the use of high-order antibody multiplexing to discriminate amongst various molecularly defined synapse types. This project will continuing efforts to make array tomography imaging methods faster, easier, more reliable and more informative and to develop high-throughput array tomographic classification tools for discrimination and quantification of synapse types. To establish a baseline quantifying normal cortical synapse populations, AT/SSA methods will be applied first to tissue specimens from wild-type mice and from autopsies and biopsies of humans unaffected by AD. The same methods will then be applied to specimens from transgenic, disease-model mice and from neurologically characterized AD patients. The information to be obtained is expected to provide new insights into the cellular and molecular basis of AD disease progression in human patients and to identify new ways that disease model mice may be used to help develop and test new drugs, vaccines, or other treatments aimed at the prevention and cure of AD. PUBLIC HEALTH RELEVANCE: The proposed research explores the potential usefulness of a new high-resolution molecular imaging technique called Array Tomography (AT) to the understanding, diagnosis, prevention, and cure of Alzheimers Disease (AD). A new set of AT-based methods will be developed and used to compare cortical synapses from the brains normal and AD-affected humans and from normal mice and mice genetically modified to mimic the human AD condition. The information to be obtained is expected to provide new insights into the cellular and molecular basis of AD disease progression in human patients and to identify new ways that disease model mice may be used to help develop and test new drugs, vaccines, or other treatments aimed at the prevention and cure of AD.