Generation and analysis of FoxG1 transgenic mouse lines

描述(申请人提供)：神经退行性疾病的特点是进行性和无情的神经元丢失。预防或减缓神经退化的一种策略是刺激具有神经保护活性的分子。利用组织培养系统，我们已经证明了FoxG1是Forkhead转录因子家族的成员，对于维持神经元的生存是必要的。此外，FoxG1的高表达具有较强的神经保护作用。我们建议扩大我们在体内的研究，并预测FoxG1水平的升高也将在神经退行性疾病的动物模型中起到保护作用。我们建议通过产生转基因小鼠来验证这一预测，转基因小鼠选择性地在神经元中表达高水平的FoxG1。这些小鼠将与两种不同的神经退行性疾病小鼠模型杂交，并评估对神经病理进展的影响。这项建议的两个具体目标是：目标1：获得过度表达野生型FoxG1的转基因小鼠，FoxG1是一种结构性活性形式的FoxG1，以及显性-阴性形式的FoxG1。我们将使用鼠普恩蛋白(MPRP)启动子来驱动这三种形式的FoxG1在小鼠中的表达。目的：分析FoxG1过表达对脑和小鼠神经退行性变模型的影响。我们将通过WT和CA FoxG1的表达来检测FoxG1活性增加对正常大脑中神经元存活的影响。在这个目标的第二部分，我们将检查FoxG1活性升高是否能保护小鼠免受神经退化的影响。我们将FoxG1高表达转基因小鼠与R6/2小鼠(HD模型)和p25/CDK5诱导转基因小鼠(AD模型)杂交。该项目将导致开发三个新的小鼠品系，表达野生型和突变型FoxG1转基因小鼠。FOXG1转基因小鼠目前还不存在。我们认为，培育这些小鼠并测试上调的FoxG1是否可以预防神经退行性疾病，将有助于深入了解FoxG1在体内有丝分裂后神经元中的功能，并可能将其确定为开发神经退行性疾病的新治疗策略的靶点。 与公共健康相关：我们实验室以前的工作已经证明，FoxG1蛋白活性的提高可以保护神经元免受死亡。我们项目的目标是通过产生表达FoxG1高活性和低活性的转基因小鼠，在体内扩展这一观察结果。然后，这些小鼠将用现有的神经退行性疾病模型进行饲养。

DESCRIPTION (provided by applicant): Neurodegenerative diseases are characterized by the progressive and relentless loss of neurons. One strategy to prevent or slow down neurodegeneration is to stimulate molecules that have neuroprotective activity. Using tissue culture systems we have demonstrated that FoxG1, a member of the Forkhead family of transcription factors, is necessary for the maintenance of neuronal survival. Furthermore, elevated expression of FoxG1 has strong neuroprotective effects. We propose to extend our studies in vivo and predict that elevated levels of FoxG1 will be also be protective in animal models of neurodegenerative disease. We propose to test this prediction by generating transgenic mice that express elevated levels of FoxG1 selectively in neurons. These mice will be crossed with two different mouse models of neurodegenerative disease and the effect on progression of neuropathology evaluated. The two specific aims of this proposal are - Aim 1: To generate transgenic mice overexpressing wild-type FoxG1, a constitutively-active form of FoxG1, and a dominant-negative form of FoxG1. We will use the mouse prion protein (mPrP) promoter to drive expression of the three forms of FoxG1 in mice. Aim 2: Analysis of the effect of FoxG1 overexpression on the brain and in mouse models of neurodegeneration. We will examine the effect of increasing FoxG1 activity, through expression of WT and CA FoxG1, on neuronal survival in the normal brain. In a second part of this aim we will examine whether elevated FoxG1 activity protects mice against neurodegeneration. We will cross FoxG1-overexpressing transgenic mice with R6/2 mice (a model of HD) and with p25/CDK5 inducible-transgenic mice (a model of AD). The project will result in the development of three new mouse lines expressing wild-type and mutant FoxG1 transgenic mice. FoxG1 transgenic mice do not currently exist. We feel that generating these mice and testing whether elevated FoxG1 can protect against neurodegenerative disease will shed insight into the function of FoxG1 in postmitotic neurons in vivo and could identify it as a target for the development of novel therapeutic strategies for neurodegenerative disorders. PUBLIC HEALTH RELEVANCE: Previous work in our laboratory has demonstrated that elevated activity of the FoxG1 protein protects neurons against death. The goal of our project is to extend this observation in vivo by generating transgenic mice that express higher and lower activity of FoxG1. These mice will then be bred with existing models of neurodegenerative disease.