Tauopathies and neuronal connectivity: diffusion imaging and CLARITY

Tau蛋白病和神经元连接：扩散成像和 CLARITY

基本信息

批准号：
9112509
负责人：
KRISTI A CLARK
金额：
$ 24.3万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9112509
关键词：
Affect Algorithms Alzheimer&apos s Disease Animal Model Area Autopsy Axon Biological Biological Models Brain Brain imaging Caliber Cells Cellular biology Clinical Protocols Companions Complex Corpus Callosum Data Detection Development Diffusion Diffusion Magnetic Resonance Imaging Disease Disease Progression Disease model Doctor of Medicine Doctor of Philosophy Early Diagnosis Formalin Future Geometry Goals Hippocampus (Brain)Human Image Immunohistochemistry Individual Interdisciplinary Study Lead Location Maps Measures Methods Microscopy Modeling Molecular Molecular Biology Monitor Nerve Degeneration Neuroanatomy Neurodegenerative Disorders Neurofibrillary Tangles Neurons Oligodendroglia Pattern Post-Translational Protein Processing Preparation Process Progressive Supranuclear Palsy Property Protocols documentation RNA Processing Resolution Sampling Sensitivity and Specificity Site Staging Structure of genu of corpus callosum System Tauopathies Technology Testing Time Tissue Model Tissue Sample Tissue imaging Tissues Variant base brain tissue disorder control extracellular frontal lobe human subject human tissue in vivo insight microscopic imaging millimeter myelination neurofilament neuroimaging neuropathology non-invasive imaging normal aging novel protein aggregate public health relevance tau Proteins tau aggregation tissue processing tool transcriptome sequencing treatment response two-photon white matter

Affect Algorithms Alzheimer&apos s Disease Animal Model Area Autopsy Axon Biological Biological Models Brain Brain imaging Caliber Cells Cellular biology Clinical Protocols Companions Complex Corpus Callosum Data Detection Development Diffusion Diffusion Magnetic Resonance Imaging Disease Disease Progression Disease model Doctor of Medicine Doctor of Philosophy Early Diagnosis Formalin Future Geometry Goals Hippocampus (Brain)Human Image Immunohistochemistry Individual Interdisciplinary Study Lead Location Maps Measures Methods Microscopy Modeling Molecular Molecular Biology Monitor Nerve Degeneration Neuroanatomy Neurodegenerative Disorders Neurofibrillary Tangles Neurons Oligodendroglia Pattern Post-Translational Protein Processing Preparation Process Progressive Supranuclear Palsy Property Protocols documentation RNA Processing Resolution Sampling Sensitivity and Specificity Site Staging Structure of genu of corpus callosum System Tauopathies Technology Testing Time Tissue Model Tissue Sample Tissue imaging Tissues Variant base brain tissue disorder control extracellular frontal lobe human subject human tissue in vivo insight microscopic imaging millimeter myelination neurofilament neuroimaging neuropathology non-invasive imaging normal aging novel protein aggregate public health relevance tau Proteins tau aggregation tissue processing tool transcriptome sequencing treatment response two-photon white matter

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项目摘要

DESCRIPTION (provided by applicant): Bridging an understanding of neuroanatomy from the cellular level of microns to the systems level of millimeters is both challenging and important at this time. We are uniquely poised to undertake this multidisciplinary research topic applied to the human postmortem brain, because we have two PI's: Dr. Carol Miller, M.D., with expertise in the neuropathology and cell and molecular biology of neurodegenerative diseases, such as tauopathies, especially Alzheimer's disease (AD), and FTLD-tau variants, using the new CLARITY preparation method for large tissue volume; and Kristi Clark, Ph.D., a neuroscientist with expertise in developing novel neuroimaging methods of high angular and spatial resolution diffusion MRI (dMRI). AIM 1 Examines tau protein detection and effects of axonal aggregates in the human postmortem corpus callosum in AD, and in disease controls, e.g. progressive supranuclear palsy (PSP) a tauopathy, and in neurologically normal, age-matched controls. First, tissues (~1cm blocks) are imaged with dMRI followed by three dimensional (3D) 2-photon microscopy imaging of CLARITY processed tissue immunostained for tau, phospho-tau and neurofilament. Then, we will spatially co-register the dMRI data with the 3D 2-photon microscopy and superimpose the maps to evaluate the sensitivity and specificity of several dMRI metrics for the detection of tau. In AIM 2, cross-sections of hippocampus, one of the earliest sites affected in AD, will be analyzed by dMRI models followed by CLARITY-processed tissues to determine effects of tau aggregation on neuronal connectivity. With the CLARITY protocol applied to formalin-fixed, hippocampal cross-sections, we have already achieved immunolabeling with neuronal subtype markers in 500μm tissue sections. Similarly, we have successfully acquired our dMRI acquisition on the ex vivo human hippocampus, achieving delineation of hippocampal sub-regions and the connectivity among the sub-regions. These studies could potentially lead to development of an in vivo, non-invasive imaging protocol for clinical use in the early diagnosis and monitoring of treatment responses in a myriad of neurodegenerative diseases where protein aggregates form. In future studies, companion, unfixed tissue samples could be examined for molecular correlates pinpointing changes in RNA processing and post-translational modifications, which cannot be done in disease model systems.

描述（由适用提供）：桥接对神经解剖学的了解从微米的细胞水平到毫米的系统水平既挑战又重要。我们很毒，可以接受这一多学科研究主题，适用于人类后大脑的大脑，因为我们有两个PI：Carol Miller，M.D。博士，具有神经病理学，细胞和分子生物学的专业知识，神经退行性疾病的分子生物学，尤其是Alzheimer's Priantiation（Alzheimer's Priantiation），尤其是Alzheimer's Priantion and Frientiant and varient for varient and fiment-TAU，FTLED-TAU。组织体积；和克里斯蒂·克拉克（Kristi Clark）博士，他是一名具有专业知识的神经科学家，以开发高角度和空间分辨率扩散MRI（DMRI）的新型神经影像学方法。 AIM 1检查了tau蛋白的检测和轴突聚集体在AD和疾病控制中的人类后call体中的轴突骨料的作用，例如进行性上腹部麻痹（PSP）一种tauopathy，在神经学正常的年龄匹配的对照中。首先，用DMRI对组织（〜1cm块）进行成像，然后对tau，磷酸-TAU和神经丝进行免疫染色的透明组织的三维（3D）2个光显微镜成像。然后，我们将在空间上与3d 2光子显微镜一起将DMRI数据共同批准，并叠加地图以评估几个DMRI指标检测TAU的灵敏度和特异性。在AIM 2中，将通过DMRI模型分析海马的横截面，这是AD中最早的部位之一，然后使用清晰处理的组织进行分析，以确定TAU聚集对神经元连接性的影响。通过适用于福尔马林固定的海马横截面的清晰度方案，我们已经在500μm组织切片中使用神经元亚型标记实现了免疫标记。同样，我们已经成功地获取了对离体海马的DMRI获取，从而实现了海马子区域的划定以及子区域之间的连通性。这些研究可能会导致在早期诊断和监测蛋白质聚集物形成的无数神经退行性疾病中的早期诊断和监测治疗反应中的临床使用方案的发展。在未来的研究中，可以检查伴侣，未固定的组织样品的分子相关性，可以确定RNA加工和翻译后修饰的变化，这在疾病模型系统中无法进行。