DEVELOPMENT OF ANIMAL MODELS OF ALZHEIMER'S DISEASE USING TRANSGENIC MICE

本项目提出的研究旨在开发一系列 阿尔茨海默病的动物模型， 转基因小鼠生产。 这种模式的价值在于， 允许关于生物和分子基础的各种假设， 在一个可重复的、遗传一致的系统中测试疾病。 目前还没有这样的制度，实行这一制度将加快 开发新的诊断工具以及新的治疗方法 治疗。 待创建的特定小鼠的选择基于 有证据表明β-淀粉样蛋白前体 蛋白质在AD的发展中 将产生四种类型的小鼠。 第一种类型将携带称为LC 99的人工构建体，其中 截短的betaAPP cDNA（缺失前导序列之间的所有编码序列）， 序列和最后99个氨基酸）由巨细胞病毒驱动 启动子 由于CMV是一种几乎没有组织特异性的强启动子， 该构建体应该混杂地产生其淀粉样蛋白生成片段 整个老鼠。 LC 99的特异性裂解产物已经被发现。 在体外进行了良好的表征， 活小鼠的组织处理类似的人工肽。 的 其它三个转基因构建体将由Thy-1启动子驱动 在我们的实验室里开发的。 该启动子驱动外源基因的表达。 在大多数中枢神经系统神经元和血管周围星形胶质细胞中的基因。 我们将使用这种组织特异性表达模式来驱动三个 i）用于与不同编码序列比较的LC 99序列， 第一个实验，ii）β APP 695 cDNA的val 717-> ile突变 和iii）野生型β APP 695。 第二个结构是 与哈代发现的突变相同， 家族性阿尔茨海默病的主要形式，并应提供一个 增加检测大脑神经病理变化的可能性 第三种结构是用来作为一种 控制的哈代突变系，使我们能够评估的影响， 突变本身。 我们将进行的分析是生物化学的， 形态计量学我们的目标是彻底分析， 定量方法，我们创建的每一个转基因品系， 避免遗漏可能细微但重要的病变， 完美的动物模型的过早声明。 三位共同调查员 该项目的专家具有互补的专业知识领域，应确保 朝着建立有用的动物模型的目标稳步前进。 老年痴呆症

The research proposed in this project is directed at developing a series of animals models for Alzheimer's disease using the technology of transgenic mouse production. The value of such a model is that it would allow various hypotheses concerning the biological and molecular bases of the disease to be tested in a reproducible, genetically uniform system. No such system exists at present and its introduction would speed the development of new diagnostic tools as well as novel therapeutic treatments. The specific mice to be created have been chosen based on evidence that points to the central role of the beta-amyloid precursor protein in the development of AD. Four types of mice will be produced. The first type will carry an artificial construct known as LC99 in which a truncated betaAPP cDNA (missing all coding sequences between the leader sequence and the last 99 amino acids) is driven by the cytomegalovirus promoter. Since CMV is a strong promoter with little tissue specificity, this construct should produce its amyloidogenic fragment promiscuously throughout the mouse. The specific cleavage products of LC99 have been well characterized in vitro and it will be informative to determine how the tissues of the live mouse deal with similar artificial peptides. The other three transgenic constructs will be driven by the Thy-1 promoter developed in our laboratory. This promoter drives expression of foreign genes in most CNS neurons and in a population of perivascular astrocytes. We will use this tissue specific pattern of expression to drive three different coding sequences: i) the LC99 sequences for comparison with the first experiment, ii) the val7l7 -> ile mutation of the betaAPP695 cDNA and iii) the wild-type form of betaAPP695. The second construct is identical to the mutation discovered by Hardy in association with a dominant form of familial Alzheimer's disease and should provide an increased likelihood of detecting neuropathological changes in the brains of the mice that carry it. The third construct is meant to serve as a control to the Hardy mutation lines, allowing us to assess the impact of the mutation itself. The analyses we will perform are biochemical and morphometric. Our goal is to thoroughly analyze, with rigorous quantitative methods, each transgenic line that we create in order to avoid missing lesions that might be subtle but important, and to prevent premature claims of a perfect animal model. The three co-investigators of this project have complementary areas of expertise that should ensure steady progress towards the goal of creating useful animal models of Alzheimer's disease.