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

本项目中提出的研究是针对开发一系列 使用阿尔茨海默病动物模型的技术 转基因小鼠的生产。这种模式的价值在于，它将 允许关于生物和分子基础的各种假说 这种疾病将在一个可复制的、基因统一的系统中进行测试。 目前还不存在这样的系统，它的引入将加快 开发新的诊断工具和新的治疗方法 治疗。要创建的特定老鼠是根据以下条件选择的 表明β-淀粉样蛋白前体的中心作用的证据 蛋白质与阿尔茨海默病的发生发展。将生产四种类型的小鼠。 第一种类型将携带一种名为LC99的人造结构，其中 截短的BetaAPP cDNA(缺少前导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 推动者。由于CMV是一个很强的启动子，几乎没有组织特异性， 这种结构应该混杂地产生它的淀粉样变性片段。 在整个老鼠身上。LC99的特定切割产物已被 在体外有很好的特性，这将是确定如何 活着的老鼠的组织处理的是类似的人造多肽。这个 其他三个转基因构建物将由Thy-1启动子驱动 这是我们实验室研发的。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群体中的基因。 我们将使用这种特定于组织的表达模式来驱动三个 不同的编码序列：i)将LC99序列与 第一个实验，II)βAPP695基因的val717-和gt；ile突变 和iii)βAPP695的野生型。第二个构造是 与哈代发现的突变完全相同，与 家族性阿尔茨海默病的主要形式，并应提供 增加检测大脑中神经病变的可能性 携带病毒的老鼠。第三个构造用于充当 控制哈代突变系，使我们能够评估 突变本身。我们将执行的分析是生化和 形态测量学。我们的目标是彻底分析，以严谨的 定量方法，我们创造的每一株转基因株系 避免遗漏可能是细微但重要的损伤，并防止 过早地宣称有一个完美的动物模型。三名联合调查员 具有互补的专业知识领域，应确保 朝着创建有用的动物模型的目标稳步前进 阿尔茨海默氏症。

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.