Transgenic Mice for the Visualization of Dopamine Neurons in vivo

描述（由申请人提供）：帕金森病（PD）是一种由大脑黑质区域内多巴胺神经元进行性变性引起的神经系统疾病。这种退化过程的原因尚不清楚，然而，许多类型的化合物，如生长因子和抗凋亡药物，已被提出防止多巴胺神经元的损失。评估这些化合物的一个主要障碍是缺乏用于高通量评估的体内模型分析系统。目前，对这些化合物的测试需要对每只动物的大脑进行数周的处理和分析，然后才能通过对黑质内多巴胺神经元的量化来确定这些化合物的效果。我们提出了一种新的方法，在黑质内的多巴胺神经元可以在单个动物中可视化和量化，通过创建表达萤火虫酶荧光素酶的转基因小鼠。这种酶的表达将被限制在体内产生酪氨酸羟化酶的细胞中，酪氨酸羟化酶是合成多巴胺的限速酶。这种限制性分布将通过使用酪氨酸羟化酶启动子来驱动荧光素酶基因的表达来实现。荧光素酶的细胞特异性表达将使我们能够可视化活小鼠黑质中量化的多巴胺神经元，并为筛选影响多巴胺能神经元变性和存活的化合物提供高通量的检测方法。公共卫生相关性：帕金森病（PD）是一种以震颤、僵硬和运动迟缓为特征的神经系统疾病。这是一种逐渐恶化的疾病，目前在美国约有150万人受到影响。目前还没有治愈这种疾病的方法。在我们的提案中，我们打算开发一种转基因小鼠，使我们能够在活体动物中可视化多巴胺神经元。使用这种动物，我们将能够以高通量的方式评估防止多巴胺神经元损失的化合物，这将使我们能够快速筛选许多可能的化合物。此外，这种转基因动物的发展将使我们能够研究可能用于产生可移植到帕金森病患者体内的多巴胺神经元的诱导线索。因此，这种转基因小鼠可能在开发治疗帕金森病的新疗法中发挥重要作用。

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is a neurological disorder caused by the progressive degeneration of dopamine neurons within the substantia nigra region of the brain. The causes of this degenerative process have yet to be clearly determined, however, many types of compounds such as growth factors, and anti-apoptotic agents have been proposed to protect against the loss of dopamine neurons. A major impediment to the assessment of these compounds is the lack of an in vivo model assay system for high through-put evaluation. Currently, the testing of these compounds requires that the brains of each animal be processed and analyzed over several weeks before the effects of these compounds can be determined by the quantification of dopamine neurons within the substantia nigra. We propose a new approach where the dopamine neurons within the substantia nigra can be visualized and quantified in individual animals be creating transgenic mice that express the firefly enzyme luciferase. The expression of this enzyme will be restricted to cells within the body that produce the enzyme tyrosine hydroxylase, the rate limiting enzyme in the synthesis of dopamine. This restricted distribution will be accomplished by using the tyrosine hydroxylase promoter to drive the expression of the luciferase gene. Cell specific expression of luciferase will enable us to visualize the quantify dopamine neurons in the substantia nigra of living mice, and provide a high through-put assay for screening compounds that affect dopaminergic neuron degeneration and survival. PUBLIC HEALTH RELEVANCE: Parkinson's disease (PD) is a neurological disorder characterized by tremor, rigidity, and bradykinesia. This is a progressively deteriorating condition that currently affects about 1.5 million people in the United States. At the present time, there is no cure for this disease. In our proposal, we intend to develop a line of transgenic mice that will enable us to visualize dopamine neurons in the living animal. Using this animal, we will be able to evaluate compounds that protect against dopamine neuron loss in a high throughput manner that will allow us to quickly screen many possible compounds. In addition, the development of this transgenic animal will allow us to study inductive cues that might be used to produce dopamine neurons that can be transplanted into patients with Parkinson's disease. As a consequence, this transgenic mouse could play an important role in developing new therapies for treating Parkinson's disease.