Automated cell-type-specific electrophysiology for understanding circuit dysregulation in Alzheimer's Disease

自动化细胞类型特异性电生理学用于了解阿尔茨海默氏病的电路失调

• 批准号: 10525870
• 负责人: Craig Forest
• 金额: $ 226.33万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-17 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10525870
• 关键词: Action Potentials; Acute; Address; Affect; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease diagnosis; Alzheimer' s disease model; Alzheimer' s disease patient; Amyloid Proteins; Animal Model; Area; Biochemical Markers; Biological Markers; Brain; Brain region; Cell Therapy; Cells; Clinical Research; Cognition; Data Set; Dementia; Deposition; Detection; Development; Diagnostic tests; Disease; Electrophysiology (science); Engineering; Epilepsy; Evolution; Fluorescence; Functional disorder; Future; Goals; Hippocampus (Brain); Human; Impaired cognition; Individual; Interneurons; Label; Lead; Learning; Memory; Memory Loss; Methods; Modeling; Molecular; Mus; Neocortex; Nerve Degeneration; Neurobiology; Neurofibrillary Tangles; Neurons; Neurosciences; Optics; Parvalbumins; Pathologic; Pathology; Persons; Physiological; Physiology; Property; Research Personnel; Resolution; Robotics; Science; Senile Plaques; Slice; Sorbus; Symptoms; Technology; Therapeutic; Time; Work; base; cell type; commercialization; effective therapy; entorhinal cortex; excitatory neuron; familial Alzheimer disease; fluorescence imaging; forest; in vivo; machine vision; mild cognitive impairment; mouse model; neuronal circuitry; neurotransmission; novel; operation; optogenetics; patch clamp; pre-clinical; prevent; progressive neurodegeneration; robotic system; success; tau Proteins; theories; tool; tool development

Project Summary Over 150 million people are projected to be living with dementia worldwide by 2050. Alzheimer’s disease (AD) is the most common form of dementia, responsible for ~70% of dementia cases. A hallmark pathological feature of Alzheimer’s disease (AD) is progressive neurodegeneration, which is thought to initiate and track progressive cognitive decline in AD patients. While increasingly sensitive biochemical markers are available to diagnose AD in individuals, treatments to stall the disease in its early stages remain elusive. Increasing evidence in AD patients models indicates that significant accumulation of these biomarkers may be preceded by early circuit dysfunction. A large plurality of AD patients display subclinical epilepsy. Furthermore, circuit hyperexcitability has been observed before plaque formation in several familial AD mouse models as well, with similar findings in mouse models of sporadic AD. Cellular evidence from these studies suggests that circuit dysregulation is due to altered circuit inhibition from GABAergic interneurons. In particular, parvalbumin-expressing (PV) interneurons appear to be prone to changes in their action potential (AP) firing, and potentially neurotransmission, across distinct familial and sporadic AD mouse models. Neurodegeneration in AD is often thought to progress through well- defined brain regions, and interestingly, hyperexcitable circuits may accelerate this pathology. Whether physiological changes to PV interneurons emerge first in regions of high vulnerability in AD is unclear. Our central hypothesis is that PV interneurons will develop dysfunctional physiological deficits first in vulnerable brain regions during early AD, which may then progress to other brain areas in a Braak-esque fashion. To evaluate this hypothesis, which will require electrophysiological recordings from thousands of individual neurons, we will use the PatcherBot, our robotic platform capable of performing high-throughput, automated electrophysiology of neurons in brain slices; however, in this work, we will augment the PatcherBot’s machine vision capabilities with fluorescence imaging to specifically target PV-expressing interneurons in brain slices. The rationale and feasibility of this proposal are shown in preliminary work, demonstrating (1) fully automated patch clamping of florescent-targeted interneurons using the PatcherBot, (2) brain-wide introduction of PV specific labeling and optogenetic methods in AD mice in vivo, and (3) early-stage PV firing and neurotransmission deficits in a prodromal FAD mouse model. Here, we will address our hypothesis across three major regions (entorhinal cortex, hippocampus, isocortex) in 3 distinct models (APOE4, hAPP-KI, 5xFAD) and 3 relevant developmental timepoints. Findings from this proposal will yield wide-ranging advances, including high-throughput cell-type- specific physiology, to information regarding the potential circuit-seeding of cognitive dysfunction in early AD.

项目摘要 预计到2050年，全球有超过1.5亿人患痴呆症。阿尔茨海默氏病（AD） 是痴呆症最常见的形式，造成约70％的痴呆病例。标志性的病理特征 阿尔茨海默氏病（AD）是进行性神经变性，被认为可以启动和跟踪渐进性 AD患者的认知能力下降。虽然越来越敏感的生化标记可用于诊断AD 在个体中，在早期阶段停滞这种疾病的治疗仍然难以捉摸。越来越多的AD患者证据 模型表明，这些生物标志物的显着积累可能是早期电路功能障碍。 大量多个AD患者表现出亚临床癫痫。此外，电路过度克服性已经 在几种家庭AD小鼠模型中也观察到牙菌斑形成之前，在小鼠中有相似的发现 零星广告的模型。这些研究的细胞证据表明，电路失调是由于改变 GABA能中间神经元的电路抑制。特别是表达白蛋白的（PV）中间神经元出现 容易改变其动作潜力（AP）发射和潜在的神经传递 家族和零星的AD小鼠模型。通常认为AD中的神经变性是通过良好的 定义的大脑区域，有趣的是，过度可见的电路可能会加速这种病理。无论 在AD高脆弱性地区，PV中间神经元的生理变化尚不清楚。我们的中心 假设是PV中间神经元将在脆弱的大脑区域中首先发展出功能失调的生理缺陷 在AD早期，它可能会以Braak风格的方式发展到其他大脑区域。评估这一点 假设将需要数千个单个神经元的电生理记录，我们将使用 Patcherbot，我们的机器人平台，能够执行高通量的自动电生理学 脑切片中的神经元；但是，在这项工作中，我们将通过 荧光成像以特异性靶向大脑切片中表达PV的中间神经元。理由和 该提案的可行性在初步工作中显示，证明（1）全自动补丁夹具的夹具 使用PatcherBot，（2）PV特定标签和 AD小鼠体内的光遗传学方法，以及（3）早期PV发射和神经传递缺陷 前驱球小鼠模型。在这里，我们将解决三个主要地区的假设（内河 3种不同的模型（APOE4，HAPP-KI，5XFAD）和3个相关的发展中的皮层，海马，等皮质） 时间点。该提案的发现将产生广泛的进步，包括高通量细胞类型 - 特定的生理学，有关早期认知功能障碍潜在电路种子的信息。