CORE--SCIENTIFIC CORE

在该计划项目中提出了多学科的方法来 定义和描述突触素家族的基础 磷蛋白促进突触的形成和稳定突触。一个更伟大的 对导致这些问题的分子机制的理解 效果应导致新的治疗靶点的治疗 阿尔茨海默氏症。广泛的研究将在不同的 组织复杂程度，以定义其营养特性 突触素，包括纯化的体外生化研究 分子，适应实验操作的细胞系统， 以及对完好动物的形态和行为研究，分析 作为目标的结果可以观察到的表型变化 小鼠突触素基因中的一个或两个缺失。 科学核心旨在提供一系列技术支持 向计划项目的其他成员提供服务。责任 将包括关键技术的生产和供应 试剂和表现例行公事，但基本任务将是 完成研究项目中所述的研究所需 申请。支助职能的集中组织将 促进以可靠、高效且经济实惠的方式确保 材料供应充足。核心员工将维持殖民地 突触素I、突触素II和突触素I/II双基因敲除小鼠和 对后代进行基因分型。动物将被培育成提供 在项目I、II和IV中研究所需的动物数量和常规 将提供计时怀孕雌性婴儿的分娩，以支持培养 设施(具体目标1)。核心将保持组织培养 提供持续供应培养的小鼠胚胎的设施 海马神经元至投射I和投射II(特定目标2)。核心意志 负责多克隆抗体的生产和筛选 项目二和项目三所需的资源，包括具体领域和 非洲爪哇突触素I和II亚型的物种选择性抗体，以及 MAP激酶依赖位点的磷酸化状态特异性抗体 关于哺乳动物突触素II和非洲爪哇突触素I和II(特定目标3)。

A multi-disciplinary approach is proposed in this Program Project to define and characterize the basis by which the synapsin family of phosphoproteins promote synaptogenesis and stabilize synapses. A greater understanding of the molecular mechanisms which are responsible for these effects should lead to novel therapeutic targets for the treatment of Alzheimer's disease. A wide range of studies will be performed at distinct levels of organizational complexity to define the trophic properties of synapsins, encompassing in vitro biochemical studies with purified molecules, cellular systems that accommodate experimental manipulation, and morphological and behavioral studies in intact animals, analyzing the phenotypic changes that can be observed as a consequence of the targeted deletion of one or both of the synapsins genes in mice. The Scientific Core is designed to provide a range of technical support services to the other members of the Program Project. The responsibilities of the Scientific Core will include the production and supply of key reagents and the performance of routine, yet essential tasks that will be required to accomplish the studies described in research Projects of the application. The centralized organization of support functions will facilitate a reliable, efficient, and cost-effective means to ensure an adequate supply of materials. The Core staff will maintain the colonies of synapsin I, synapsin II, and synapsin I/II "double" knock-out mice and perform genotyping of offspring. Animals will be bred to provide the required number of animals for study in Projects I, II and IV, and regular delivery of timed-pregnant females will be provided to support the culture facility (Specific Aim 1). The Core will maintain a tissue culture facility to provide a continuous supply of cultured mouse embryonic hippocampal neurons to Projects I and II (Specific Aim 2). The Core will be responsible for the production and screening of polyclonal antibodies that are required for Projects II and III, including domain-specific and species-selective antibodies to Xenopus synapsin I and II isoform, and phosphorylation state-specific antibodies for MAP kinase-dependent sites on mammalian synapsin II and Xenopus synapsins I and II (Specific Aim 3).