Array Tomographic Single-Synapse Analysis of Normal and Disordered Cortex

描述(由研究人员提供)：这项拟议的研究探索了一种新的高分辨率组织分子成像方法-阵列断层成像(AT)对阿尔茨海默病(AD)的理解、诊断、预防和治疗的有用性。作为这项工作的一部分，将开发一套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)，目的是对小鼠和人大脑和海马皮层组织中的单个突触进行量化和分类。皮质突触丢失被广泛认为是AD和许多其他神经退行性疾病认知功能障碍的近端原因，但由于现有方法的技术限制，关于这种皮质突触缺失的定量信息有限。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稀缺。这些缺陷是人类心理健康的结果，因为强有力的证据表明，不同神经化学定义的突触类型在AD中表现出不同的病理，因此代表着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，能够分辨和定量三维体积的皮质神经管中的单个突触点状突触，并使用高阶抗体多路复用来区分各种分子定义的突触类型。该项目将继续努力使阵列断层成像方法更快、更容易、更可靠和提供更多信息，并开发用于区分和量化突触类型的高通量阵列断层成像分类工具。为了建立一个量化正常皮质突触种群的基线，AT/SSA方法将首先应用于野生型小鼠的组织样本以及未受AD影响的人类的尸检和活组织检查。然后，同样的方法将应用于转基因疾病模型小鼠和具有神经学特征的AD患者的样本。预计将获得的信息将为人类患者AD疾病进展的细胞和分子基础提供新的见解，并确定疾病模型鼠可能被用于帮助开发和测试旨在预防和治疗AD的新药、疫苗或其他治疗方法的新方法。公共卫生相关性：这项拟议的研究探索了一种名为阵列断层扫描(AT)的新的高分辨率分子成像技术对阿尔茨海默病(AD)的理解、诊断、预防和治疗的潜在用处。将开发一套新的基于AT的方法，并用于比较大脑正常和受AD影响的人以及正常小鼠和经过基因改造以模拟人类AD条件的小鼠的皮质突触。预计将获得的信息将为人类患者AD疾病进展的细胞和分子基础提供新的见解，并确定疾病模型鼠可能被用于帮助开发和测试旨在预防和治疗AD的新药、疫苗或其他治疗方法的新方法。

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.