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: 6872660
• 负责人: TONY WYSS-CORAY
• 金额: $ 14.56万
• 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/6872660
• 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可导致中风或神经退行性疾病中的细胞死亡和退化。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的信息或研究致病途径。这与许多外周疾病形成鲜明对比，在许多外周疾病中，可以更容易地测量血液中的疾病标志物，或者可以获得活组织检查，而不会显着干扰疾病。因此，需要新的方法来无创地研究大脑中的疾病进展或细胞变化。活体动物的生物发光成像最近已被用于监测和定量外周器官的基因活性和疾病进展，并取得了巨大成功。虽然这种成像方式缺乏高分辨率，目前不能用于高精度定位信号，但它是定量的，如果使用适当的融合基因构建体，可以如实地报告基因激活。在这里，我们建议在含有损伤反应性荧光素报告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量。强有力的初步数据表明，我们确实可以从活体小鼠受伤的大脑中收集与报告基因活性相关的光子。我们建议使用这些携带对 TGF-β 信号通路有反应的报告基因的小鼠来优化大脑的非侵入性生物发光成像，并尝试使用生物发光在两种急性脑损伤模型中非侵入性地定量神经元损伤成像（具体目标 1}。我们还建议设计相关小鼠，使其在特定的神经元组中高水平表达荧光素酶和 YFP，以尝试将活体小鼠的生物发光信号与神经元细胞数量相关联（具体目标 2）如果我们成功，我们的研究将证明使用非侵入性生物发光成像来测量大脑中的基因活性并将此类测量结果用作个体小鼠的病理或生理过程的指标的可行性。报告小鼠还可用于筛选药物以干扰或激活活体小鼠中特定信号通路的功效，并评估药物在大脑中的可用性。