Genetic labeling and visualization of CSMN in models of motor neuron disorders

运动神经元疾病模型中 CSMN 的基因标记和可视化

• 批准号: 8623379
• 负责人: Pembe Hande Ozdinler
• 金额: $ 23.18万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8623379
• 关键词: ARHGEF5 gene; Affect; Amyotrophic Lateral Sclerosis; Anatomy; Attention; Biology; Brain; CCL1 gene; Cellular Structures; Cellular biology; Cerebral cortex; Cessation of life; Complex; Corticospinal Tracts; Crossbreeding; Defect; Development; Disease; EP300 gene; Frontotemporal Dementia; Frustration; Generations; Genes; Genetic; Goals; Grant; Health; Hereditary Spastic Paraplegia; Human; Human Pathology; Imagery; Knockout Mice; Label; Lead; Life; Mediating; Mind; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Motor; Motor Cortex; Motor Neuron Disease; Motor Neurons; Movement; Movement Disorders; Mus; Mutation; Neurodegenerative Disorders; Neurons; Operative Surgical Procedures; Pathology; Patients; Persons; Population; Primary Lateral Sclerosis; Reporter; Signal Transduction; Source; Spinal Cord; Staging; Structure; System; Time; Transgenic Organisms; Translating; UCHL1 gene; Visual; base; effective therapy; hippocampal pyramidal neuron; human TFRC protein; improved; in vivo; molecular marker; motor neuron degeneration; motor neuron function; mouse model; novel; promoter; protein TDP-43; public health relevance; ranpirnase; success; tool; treatment strategy; tyrosine kinase ABL1

There are thousands of different neuron populations in our cerebral cortex, but in neurodegenerative diseases only a select neuron population show primary vulnerability and undergo progressive degeneration. Corticospinal motor neurons (CSMN) are large pyramidal neurons that are located in layer V of the motor cortex. Their cellular structure is very unique and they function as the "spokes person"of the cerebral cortex for the initiation and modulation of movement. Voluntary movement is the act of a clever and well-informed mind. Therefore CSMN receive information from numerous neurons, including long-distance projection and local circuitry neurons. CSMN's unique ability to integrate and translate this information into one signal towards spinal cord targets sets it apart from other cortical neurons. Therefore its degeneration has severe consequences that lead to various movement disorders. There is a developing need to understand the basis of CSMN degeneration in diseases. However, identification and visualization of CSMN is not easy as they are embedded among thousands of other neuron populations within the cerebral cortex. We recently generated a novel reporter line, the UCHL1-eGFP mice, in which CSMN are genetically labeled in the motor cortex. eGFP expression under the control of UCHL1 promoter is stable, persistent up to P800 in vivo, and is restricted to CSMN in the motor cortex. CSMN identity of eGFP+ neurons in the motor cortex are identified by anatomy, retrograde labeling, molecular marker expression profile and electrophysiological analysis. This reporter line offers many unique advantages; a) we can for the first time visualize CSMN without any need for a retrograde labeling surgery; b) CSMN can be purified by FACS- mediated approaches at different stages in life; c) the cellular and molecular mechanisms that are responsible for CSMN vulnerability and degeneration can be studied in detail and with precision; d) most importantly this novel reporter line can be crossed to various mouse models of movement disorders to investigate the biology of CSMN with respect to disease. In this proposal, our goal is to bring visual clarity to CSMN in various mouse models of motor neuron diseases. We will be crossing UCHL1-eGFP mice with the recently identified mouse models that show potential involvement of upper motor neuron degeneration in disease pathology. Due to time limitations of R21 grant, we will characterize the timing and extent of CSMN degeneration in a limited number of mouse models, such as the Tdp43A315T and Alsin KO mice. The tools we generate and the approach we develop will help generation and characterization of other reporter mouse models, and will improve our efforts of understanding the cellular and molecular mechanisms behind CSMN vulnerability and degeneration.

在我们的大脑皮层中有成千上万种不同的神经元群体，但在神经退行性疾病中， 只有选定的神经元群体显示出原发性脆弱性并经历进行性退化。皮质脊髓 运动神经元（CSMN）是位于运动皮层的V层中的大锥体神经元。它们的细胞 结构是非常独特的，他们的功能是作为“辐条人“的大脑皮层的启动和 运动的调制。自愿的运动是一个聪明和见多识广的头脑的行为。CSMN 从许多神经元接收信息，包括长距离投射和局部电路神经元。 CSMN的独特能力，整合和翻译这些信息成一个信号对脊髓的目标设置它 与其他皮层神经元分开。因此，它的退化具有严重的后果，导致各种 运动障碍有一个发展的需要，以了解疾病的CSMN变性的基础。 然而，CSMN的识别和可视化并不容易，因为它们嵌入在数千个其他 大脑皮层内的神经元群体。我们最近产生了一种新的报告细胞系，UCHL 1-eGFP 小鼠，其中CSMN在运动皮层中被遗传标记。UCHL 1控制下的eGFP表达 该启动子在体内是稳定的，持续到P800，并且仅限于运动皮层中的CSMN。CSMN身份 通过解剖学、逆行标记、分子标记物表达鉴定运动皮质中的eGFP+神经元 轮廓和电生理分析。这条记者热线提供了许多独特的优势; a）我们可以为第一个 时间可视化CSMN，而不需要任何逆行标记手术; B）CSMN可以通过FACS纯化， 在生命的不同阶段介导的方法; c）负责的细胞和分子机制 CSMN的脆弱性和退化可以详细和精确地研究; d）最重要的是， 报告细胞系可以与各种运动障碍的小鼠模型杂交以研究CSMN的生物学 关于疾病。在这个提议中，我们的目标是在各种小鼠模型中为CSMN带来视觉清晰度。 运动神经元疾病我们将UCHL 1-eGFP小鼠与最近鉴定的小鼠模型杂交， 显示在疾病病理学中可能涉及上运动神经元变性。由于时间限制， R21授权，我们将在有限数量的小鼠中描述CSMN变性的时间和程度。 模型，如Tdp 43 A315 T和Alsin KO小鼠。我们开发的工具和方法将 帮助其他报告小鼠模型的生成和表征，并将改善我们的努力， 了解CSMN脆弱性和变性背后的细胞和分子机制。