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.

项目概要 预计到 2050 年，全球将有超过 1.5 亿人患有痴呆症。 阿尔茨海默病 (AD) 是最常见的痴呆症，约占痴呆症病例的 70%。标志性病理特征 阿尔茨海默病 (AD) 的病因是进行性神经退行性变，人们认为它会启动并追踪进行性神经退行性变。 AD患者的认知能力下降。虽然越来越敏感的生化标记物可用于诊断 AD 对于个体来说，在疾病早期阶段阻止疾病的治疗方法仍然难以捉摸。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先于早期回路功能障碍。 大量 AD 患者表现出亚临床癫痫。此外，电路过度兴奋已 在几种家族性 AD 小鼠模型中，在斑块形成之前也观察到这一现象，在小鼠中也有类似的发现 散发性 AD 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，表达小清蛋白（PV）的中间神经元出现 不同的动作电位（AP）放电和潜在的神经传递容易发生变化 家族性和散发性 AD 小鼠模型。 AD 中的神经变性通常被认为是通过良好的进展 明确的大脑区域，有趣的是，过度兴奋的电路可能会加速这种病理学。无论 PV 中间神经元的生理变化首先出现在 AD 的高度脆弱区域尚不清楚。我们的中央 假设PV中间神经元首先会在脆弱的大脑区域产生功能失调的生理缺陷 在AD早期，它可能会以布拉克式的方​​式进展到其他大脑区域。为了评价这一点 假设，这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、自动化的电生理学 大脑切片中的神经元；然而，在这项工作中，我们将通过以下方式增强 PatcherBot 的机器视觉功能： 荧光成像专门针对脑切片中表达 PV 的中间神经元。理由和 该提案的可行性在初步工作中得到了证明，证明了（1）全自动膜片钳 使用 PatcherBot 以荧光为目标的中间神经元，(2) 在全脑范围内引入 PV 特异性标记和 AD小鼠体内的光遗传学方法，以及（3）早期PV放电和神经传递缺陷 前驱期 FAD 小鼠模型。在这里，我们将在三个主要区域（内嗅区 皮层、海马体、同皮质）的 3 种不同模型（APOE4、hAPP-KI、5xFAD）和 3 种相关的发育模型 时间点。该提案的研究结果将产生广泛的进步，包括高通量细胞类型 具体的生理学，有关早期 AD 认知功能障碍的潜在电路播种的信息。