CORE--SCIENTIFIC CORE

核心--科学核心

• 批准号: 6413585
• 负责人: ANDREW J CZERNIK
• 金额: $ 24.29万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6413585
• 关键词: animal breeding; animal colony; antibody; biomedical facility; immunological substance; immunoprecipitation; laboratory mouse; reagent /indicator; synapsins; tissue /cell culture; western blottings

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).

本计划项目提出了一种多学科方法， 定义和表征突触蛋白家族的基础， 磷蛋白促进突触发生并稳定突触。更大 了解这些分子机制， 这些作用应该导致新的治疗靶点，用于治疗 老年痴呆症将在不同的地点进行广泛的研究。 组织的复杂性水平，以确定营养属性的 突触蛋白，包括在体外生物化学研究与纯化 分子，细胞系统，以适应实验操作， 以及完整动物的形态学和行为学研究， 表型变化，可以观察到的结果，靶向 在小鼠中缺失一个或两个突触蛋白基因。 科学核心旨在提供一系列技术支持 为项目的其他成员提供服务。的责任 科学核心将包括生产和供应关键 试剂和常规的性能，但必不可少的任务， 所需完成的研究项目中所描述的研究 应用程序.支持职能的集中组织将 提供可靠、高效和具有成本效益的手段，以确保 充足的材料供应。核心工作人员将维持殖民地的 突触蛋白I、突触蛋白II和突触蛋白I/II“双”敲除小鼠， 对后代进行基因分型。动物将被饲养， 项目I、II和IV中研究所需的动物数量， 将提供定时妊娠雌性动物的分娩，以支持培养 具体目标（1）。核心将维持组织培养 提供持续供应培养的小鼠胚胎的设施 海马神经元到项目I和II（具体目标2）。芯会 负责多克隆抗体的生产和筛选 项目二和项目三所需的，包括特定领域和 针对爪蟾突触蛋白I和II同种型的物种选择性抗体，和 MAP激酶依赖位点的磷酸化状态特异性抗体 哺乳动物突触蛋白II和非洲爪蟾突触蛋白I和II（具体目标3）。