MODELS OF GLUTAMATE AND NICOTINE RECEPTOR DISFUNCTION

这项提议的目的是测试基因治疗的策略 活着。这将通过在鼠标中进行工程设计来实现 两种主要兴奋性受体系统的基因组突变 哺乳动物的大脑负责调节快速突触传递， 谷氨酸受体和胆碱能烟碱受体系统。 谷氨酸受体系统介导了绝大多数的FAST 哺乳动物大脑中的突触传递，并与 记忆形成和神经退行性变的许多实验 阿尔茨海默病是在S患者中观察到的疾病。胆碱能 尼古丁受体系统与记忆功能有关 而尼古丁受体的数量在 阿尔茨海默病~S病。一种假设是尼古丁的缺陷 受体系统与观察到的记忆缺陷有关 阿尔茨海默病~S患者将接受检测。基因治疗策略将 然后在突变小鼠身上尝试恢复基因表达 和认知功能。这些实验将利用 重组DNA技术与突变小鼠的培育 通过基因工程的方法。基因打靶方法将是 用于敲除或干扰谷氨酸受体和烟碱 利用最近开发的新技术在成年小鼠中的受体基因 Cre/loxP系统。突变将被引入谷氨酸 受体基因可以改变谷氨酸受体，使它们 在过程中，异常高的钙流入细胞 正常的突触传输。将对突变的小鼠进行分析 使用各种技术。大脑将被切开并 检查大体形态变化和细胞证据 死亡。包括莫里斯迷宫在内的行为测试将用于 测试学习和记忆功能。在突变的小鼠 以神经退行性疾病和认知缺陷为特征， 治愈突变小鼠的尝试将通过表达 多种潜在的~保护性~或替代蛋白 小鼠脑内使用多个不同的病毒表达和细胞 嫁接系统。

The aim of this proposal is to test strategies for gene therapy in vivo. This will be accomplished by engineering into the mouse genome mutations in the two major excitatory receptor systems in the mammalian brain which mediate fast synaptic transmission, the glutamate receptor and the cholinergic nicotinic receptor system. The glutamate receptor system mediates the vast majority of fast synaptic transmission in the mammalian brain and is implicated by many experiments in memory formation and neurodegenerative disease as is observed in Alzheimer~s patients. The cholinergic nicotinic receptor system has been implicated in memory function and the number of nicotinic receptors is dramatically reduced in Alzheimer~s disease. The hypothesis that a defect in the nicotinic receptor system contributes to the memory deficit observed in Alzheimer~s patients will be tested. Gene therapy strategies will then be attempted in the mutant mice to restore gene expression and cognitive function. The experiments will make use of recombinant DNA technology and the production of mutant mice by genetic engineering methods. Gene targeting methods will be used to knockout or disrupt glutamate receptor and nicotinic receptor genes in the adult mouse using the recently developed CRE/LoxP system. Mutations will be introduced into glutamate receptor genes which alter the glutamate receptors so that they flux abnormally high amounts of calcium into the cell during normal synaptic transmission. The mutant mice will be analyzed using a variety of techniques. The brains will be sectioned and examined for gross morphological changes and evidence of cell death. Behavioral tests including the Morris maze will be used to test for learning and memory function. After the mutant mice are characterized for neurodegenerative disease and cognitive deficits, attempts to cure the mutant mice will be carried out by expressing various potential ~protective~ or replacement proteins in the mouse brain using a number of different viral expression and cell graft systems.