Characterization of the involvement of the cerebellum in animal models of C9orf72 ALS/FTD

C9orf72 ALS/FTD 动物模型中小脑参与的表征

• 批准号: 10041549
• 负责人: YUQING LI
• 金额: $ 41.94万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10041549
• 关键词: Acute; Affect; Age-Months; Amyotrophic Lateral Sclerosis; Anatomy; Animal Disease Models; Animal Model; Antisense RNA; Autopsy; Bacterial Artificial Chromosomes; Behavior; Behavioral; Biochemical; Biochemistry; Brain; C9ORF72; Calcium; Caregivers; Cells; Cerebellar Diseases; Cerebellar degeneration; Cerebellum; Cognitive deficits; Data; Dementia; Dipeptides; Disease; Economic Burden; Electrophysiology (science); Environment; Equilibrium; Feedback; Frontotemporal Dementia; Functional disorder; Funding; Genes; Genetic; Goals; Individual; Introns; Ion Channel; Knockout Mice; Knowledge; Language; Length; Mediating; Membrane; Messenger RNA; Molecular Genetics; Morphology; Motor; Motor Neurons; Motor output; Mus; Musculoskeletal Equilibrium; Mutant Strains Mice; Neurodegenerative Disorders; Neurological Models; Neurons; Other Genetics; Outcomes Research; Pathogenesis; Pathologic; Pathology Report; Patients; Persons; Phenotype; Play; Potassium; Potassium Channel; Property; Proteins; Purkinje Cells; RNA; Reporting; Research; Research Personnel; Resources; Role; Spinal Cord; Symptoms; Techniques; Temporal Lobe; Testing; Transcript; Transgenic Mice; Translations; United States National Institutes of Health; Walking; Western Blotting; Work; behavior test; behavioral variant frontotemporal dementia; cognitive function; early onset; effective therapy; emotional functioning; frontal lobe; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; gray matter; imaging study; interdisciplinary approach; loss of function; motor deficit; motor learning; mouse model; nervous system disorder; protein TDP-43; therapeutic target; Acute; Affect; Age-Months; Amyotrophic Lateral Sclerosis; Anatomy; Animal Disease Models; Animal Model; Antisense RNA; Autopsy; Bacterial Artificial Chromosomes; Behavior; Behavioral; Biochemical; Biochemistry; Brain; C9ORF72; Calcium; Caregivers; Cells; Cerebellar Diseases; Cerebellar degeneration; Cerebellum; Cognitive deficits; Data; Dementia; Dipeptides; Disease; Economic Burden; Electrophysiology (science); Environment; Equilibrium; Feedback; Frontotemporal Dementia; Functional disorder; Funding; Genes; Genetic; Goals; Individual; Introns; Ion Channel; Knockout Mice; Knowledge; Language; Length; Mediating; Membrane; Messenger RNA; Molecular Genetics; Morphology; Motor; Motor Neurons; Motor output; Mus; Musculoskeletal Equilibrium; Mutant Strains Mice; Neurodegenerative Disorders; Neurological Models; Neurons; Other Genetics; Outcomes Research; Pathogenesis; Pathologic; Pathology Report; Patients; Persons; Phenotype; Play; Potassium; Potassium Channel; Property; Proteins; Purkinje Cells; RNA; Reporting; Research; Research Personnel; Resources; Role; Spinal Cord; Symptoms; Techniques; Temporal Lobe; Testing; Transcript; Transgenic Mice; Translations; United States National Institutes of Health; Walking; Western Blotting; Work; behavior test; behavioral variant frontotemporal dementia; cognitive function; early onset; effective therapy; emotional functioning; frontal lobe; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; gray matter; imaging study; interdisciplinary approach; loss of function; motor deficit; motor learning; mouse model; nervous system disorder; protein TDP-43; therapeutic target

Project Summary: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the progressive loss of motor neurons in the brain and spinal cord. Frontotemporal dementia (FTD) is early-onset dementia caused by degeneration of the frontal and temporal lobes with cognitive deficits and behavioral and language abnormalities. FTD shares pathophysiological features with ALS. A hexanucleotide repeat expansion in an intron of C9orf72 is the most frequently reported genetic cause of both diseases. Most of the pathophysiological studies of C9orf72 ALS/FTD and other types of ALS so far have been focused on upper and lower motor neurons. Recently, several functional studies have detected lower gray matter volume in the cerebellum in pre-symptomatic C9orf72 repeat expansion carriers, and anatomical evidence in C9orf72 disease mouse models have demonstrated cerebellar degeneration. Furthermore, pathology reports using postmortem or patient cells have demonstrated inclusions, RNA foci, dipeptide repeat proteins, and decreased C9orf72 mRNA and protein levels in the cerebellum, especially Purkinje cells, implicating cerebellar dysfunction in the pathogenesis of C9orf72 ALS/FTD. However, there are very few studies in animal disease models focusing on the cerebellum. Our strong preliminary data on C9orf72 knockout mice has revealed motor deficits and altered Purkinje cell activity. The functional significance of the cerebellum in the C9orf72 ALS/FTD pathogenesis is not known, let alone the detailed mechanisms. These unknowns hamper efforts to understand the pathophysiology of C9orf72 ALS/FTD and to develop effective therapies for patients. The overall goal of our research is to use transgenic mice to study the pathophysiology of C9orf72 ALS/FTD. The specific goal of this application is to characterize mouse models of C9orf72 ALS/FTD to determine the role of cerebellum in the pathogenesis of ALS and FTD. We hypothesize that at the presymptomatic stage, the C9orf72 mutant mice have motor deficits in coordination and balance, anatomical and functional deficits in the cerebellum, especially in the Purkinje cells due to the loss of function of C9orf72 protein and changes in the ion channels. We plan to test our hypothesis with the following Specific Aims: (1) we will examine C9orf72 mutant mice in the behavioral test battery, (2) we will characterize neuronal morphology, passive membrane properties, and intrinsic excitabilities of Purkinje cells, and (3) we will perform acutely dissociated Purkinje cell recording and Western blot analysis to determine ion channels involved in the altered firing of Purkinje cells. The successful completion of the Aims will expand the current knowledge concerning the involvement of the cerebellum in C9orf72-mediated ALS/FTD. This should significantly increase our understanding of the pathophysiology of C9orf72 ALS/FTD and contribute to the pathophysiological studies of other genetic ALS. Furthermore, the outcome of the research will support cerebellum as a potential therapeutic target to treat C9orf72 ALS/FTD. .

项目摘要： 肌萎缩性侧索硬化症（ALS）是一种神经退行性疾病，其特征是进行性丧失 大脑和脊髓中的运动神经元。额颞痴呆（FTD）是早期发作的痴呆 通过具有认知缺陷以及行为和语言的额叶和颞叶的变性 异常。 FTD与ALS共享病理生理特征。六核苷酸重复膨胀 C9ORF72的内含子是两种疾病的遗传原因。大多数 到目前为止，C9orf72 ALS/FTD和其他类型的ALS的病理生理研究集中在上部 和降低运动神经元。最近，一些功能研究发现了较低的灰质体积 症状前C9orf72重复膨胀载体和C9orf72的解剖学证据的小脑 疾病小鼠模型表明小脑变性。此外，病理学报告 验尸或患者细胞表现出夹杂物，RNA焦点，二肽重复蛋白，并降低 小脑，尤其是Purkinje细胞中的C9ORF72 mRNA和蛋白质水平，暗示小脑 C9orf72 ALS/FTD发病机理的功能障碍。但是，动物疾病的研究很少 专注于小脑的模型。我们关于C9orf72敲除小鼠的强大初步数据揭示了电动机 缺陷和Purkinje细胞活性改变。小脑在C9ORF72 ALS/FTD中的功能意义 发病机理尚不清楚，更不用说详细的机制了。这些未知数阻碍了理解的努力 C9ORF72 ALS/FTD的病理生理，并为患者开发有效的疗法。总体目标 我们的研究是使用转基因小鼠研究C9ORF72 ALS/FTD的病理生理。的具体目标 该应用是为了表征C9orf72 ALS/FTD的鼠标模型，以确定小脑在 ALS和FTD的发病机理。我们假设在症状阶段，c9orf72突变体 小鼠在协调和平衡，小脑中的解剖和功能缺陷方面缺乏运动缺陷， 特别是由于C9ORF72蛋白功能的损失以及离子通道的变化，尤其是在Purkinje细胞中。 我们计划以以下特定目的测试我们的假设：（1）我们将检查C9orf72突变小鼠 行为测试电池，（2）我们将表征神经元形态，被动膜特性和 Purkinje细胞的内在兴奋性，（3）我们将执行急性解离的Purkinje细胞记录和 蛋白质印迹分析以确定涉及Purkinje细胞发射改变的离子通道。成功 目的的完成将扩大有关小脑参与的当前知识 C9orf72介导的ALS/FTD。这应该大大提高我们对 C9orf72 ALS/FTD，并为其他遗传ALS的病理生理研究做出了贡献。此外， 该研究的结果将支持小脑作为治疗C9orf72 ALS/FTD的潜在治疗靶标。 。