Live Imaging of Neuronal Injury in Reporter Mice

报告小鼠神经元损伤的实时成像

• 批准号: 7005430
• 负责人: TONY WYSS-CORAY
• 金额: $ 10.41万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-15 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7005430
• 关键词: bioimaging /biomedical imaging; biological signal transduction; bioluminescence; biomarker; brain imaging /visualization /scanning; brain metabolism; cellular pathology; charge coupled device camera; disease /disorder model; gene expression; genetically modified animals; histology; laboratory mouse; luciferin monooxygenase; nerve injury; nervous system disorder diagnosis; neural degeneration; noninvasive diagnosis; reporter genes; transforming growth factors

Acute or chronic neuronal injury can lead to cell death and degeneration in stroke or neurodegenerative diseases. A number of animal models have been developed to study these diseases but in most cases, animals have to be sacrificed to gain information on disease severity or to study pathogenic pathways. This is in contrast to many peripheral diseases where disease markers can be more easily measured in the blood or biopsies can be obtained without interfering dramatically with the disease. Therefore, new methods are necessary to study disease progression or cellular changes in the brain noninvasively. Bioluminescence imaging in living animals has recently been used to monitor and quantitate gene activity and disease progression in peripheral organs with great success. Although, this imaging modality lacks high resolution and cannot be used at present to localize signals with high accuracy, it is quantitative and can faithfully report gene activation if appropriate fusion gene constructs are used. Here we propose to use bioluminescence imaging in transgenic mice that harbor an injury-responsive Lucifer's reporter gene to assess neuronal injury or use mice that express luciferase at high levels in neurons to quantitate cell number. Strong preliminary data indicate that we can indeed collect photons that correlate with reporter gene activity from injured brains of living mice. We propose to use these mice, which carry a reporter gene responsive to the TGF-beta signaling pathway, to optimize non-invasive bioluminescence imaging of the brain and to try to quantitate neuronal injury non-invasively in two acute models of brain injury using bioluminescence imaging (Specific Aim 1}. We also propose to engineer related mice that express luciferase and YFP constitutively at high levels in defined groups of neurons to try to correlate bioluminescence signals in living mice with neuronal cell number (Specific Aim 2). If we are successful, our studies will demonstrate feasibility of using non-invasive bioluminescence imaging to measure gene activity in the brain and to use such measurements as indicators of pathological or physiological processes in individual mice. These, and other reporter mice could also be used to screen drugs for efficacy to interfere with or activate specific signaling pathways in living mice and to assess drug availability in the brain.

急性或慢性神经元损伤会导致中风或神经退行性疾病的细胞死亡和变性。已经开发了许多动物模型来研究这些疾病，但在大多数情况下，必须牺牲动物才能获得有关疾病严重程度或研究致病途径的信息。这与许多周围疾病相反，在没有剧烈干扰该疾病的情况下，可以在血液或活检中更容易地测量疾病标志物。因此，对于研究大脑的疾病进展或细胞变化是无创的。最近，活跃动物中的生物发光成像已用于监测和定量基因活性和疾病进展，并取得了巨大成功。尽管这种成像方式缺乏高分辨率，目前不能使用高精度来定位信号，但它是定量的，如果使用适当的融合基因构建体，则可以忠实地报告基因激活。在这里，我们建议在具有损伤响应性路西法的记者基因的转基因小鼠中使用生物发光成像来评估神经元损伤或使用在神经元中高水平表达荧光素酶的小鼠来定量细胞数。强有力的初步数据表明，我们确实可以收集与活小鼠受伤大脑的报告基因活性相关的光子。我们建议使用这些小鼠，这些小鼠具有对TGF-beta信号通路有反应的报告基因，以优化大脑的非侵入性生物发光成像，并试图在两个急性的脑损伤中使用双应发光的脑损伤在两个急性的脑损伤中非侵入性地进行神经元损伤，并提出了与特定的AIM级相关的相关级别（我们对YF的高度相关型）。定义的神经元试图将活小鼠中的生物发光信号与神经元的细胞数相关联（特定的目的2），我们的研究将证明使用无创的生物发光成像来测量大脑中的基因活性并将这些测量作为病理学的指标。干扰或激活活小鼠中的特定信号通路并评估大脑中的药物可用性。