SCEINTIFIC CORE

阿尔茨海默病的神经病理改变及其分子和细胞的研究 它们背后的变化已经产生了关于这种毁灭性疾病的病因的丰富信息 疾病。项目1-3中概述的综合研究将在现有知识的基础上扩展 神经细胞的严格细胞生物学、分子、生化、行为和电生理学研究 在体外和几种阿尔茨海默病小鼠模型中。这些研究有可能为更深入地了解 AD的病因以及阐明AD治疗的可能新靶点。科学核心将是一个 中心致力于促进项目1-3中提议的实验，这些实验需要使用初级 神经元和器官培养，转基因动物，免疫试剂和酵母- 重组的γ-分泌酶。这个集中化设施的存在将确保 项目1-3将以最及时和最具成本效益的方式完成。许多研究都是在 该计划项目拨款将需要使用高质量的神经元培养来验证最初的发现 从转化的细胞系中。在具体目标一中，核心将负责执行例行任务 参与项目1-3所需的原代神经元和器官培养的准备工作。作为所有人 项目1-3的发现最终将在完整的动物身上进行验证和测试，实验 这些项目中概述的还将需要培育和维护转基因动物。 因此，转基因小鼠的培育和维护是核心的具体目标II。为了帮助 新的蛋白质相互作用、蛋白质定位和蛋白质磷酸化的特征， Science Core将成为新的多克隆抗体的关键来源，包括磷酸化状态- 特定目的III中描述的特定抗体。最后，Y-分泌酶活性的重建 简单的真核细胞巴斯德毕赤酵母的遗传操作和筛选是专门的目的四。

Studies of the neuropathological alterations seen in Alzheimer's Disease (AD) and the molecular and cellular changes underlying them have yielded a wealth of information regarding the etiology of this devastating disease. The combined studies outlined in Projects 1-3 will extend upon this existing knowledge with rigorous cell biological, molecular, biochemical, behavioral, and electrophysiological studies of neuronal cells in vitro and in several mouse models of AD. These studies have the potential to provide greater insight into the etiology of AD as well as to elucidate possible novel targets for AD therapy. The Scientific Core will be a center devoted to facilitating the experiments proposed in Projects 1-3 that require the use of primary neuronal and organotypic cultures, genetically modified animals, immunological reagents, and yeast- reconstituted y-secretase. The existence of this centralized facility will ensure that the experiments outlined in Projects 1-3 will be completed in the most timely and cost-effective manner. Many of the studies proposed in this Program Project Grant will require the use of high-quality neuronal cultures to validate initial findings from transformed cell lines. In Specific Aim I, the Core will be responsible for performing the routine tasks involved in the preparation of primary neuronal and organotypic cultures needed by Projects 1-3. As all of the findings of Projects 1-3 will ultimately be validated and tested in intact animals, the experiments outlined in these Projects will also require the breeding and maintenance of genetically modified animals. Thus the breeding and maintenance of transgenic mice is Specific Aim II of the Core. In order to aid in the characterization of new protein-protein interactions, protein localization, and protein phosphorylation, the Scientific Core will be a keysource of new polyclonal antibodies, including phosphorylation state- specific antibodies, as described in Specific Aim III. Finally, the reconstitution of y-secretaseactivity in the simple eukaryote P. pastoris for genetic manipulation and screening is Specific Aim IV.