ANIMAL MODELS, ELECTRON MICROSCOPY, CELL CULTURE AND MOLECULAR BIOLOGY

该计划中的所有项目都利用动物模型和细胞培养系统来研究 衰老中钙离子调节失调、氧化应激和慢性兴奋性毒性的研究。动物模型，电子 显微复制、细胞培养和分子生物学核心是一组不同的 个别实验室无法提供的资源、服务和专业知识。核心将维护网络 可访问的关系数据库，用于跟踪程序中的每一只动物并持续更新数据 用每种动物和细胞培养实验获得。该网站综合了 核心提供多样化的服务，并极大地促进所有参与者之间的信息共享。这个 以NIA群体中的F344BNF1大鼠为体内生物衰老模型。此外，还有一部小说 在神经元中过度表达谷氨酸合成酶GLUD1的转基因小鼠提供了一个模型 过量的谷氨酸释放会导致大脑的退行性变化和寿命的减弱。的工作人员 核心C维持小鼠群体，包括基因分型、杂交育种和长寿监测。 与动物系统相关的细胞培养模型在核心准备和维护，以允许 对来自完整动物研究的观察结果进行的机械测试。细胞模型包括C2C12肌细胞， 原代骨骼肌细胞、胚胎F344BNF1大鼠和GLUD1/wt小鼠的原代神经元，以及 神经元性SH-SY5Y细胞株。研究人员将在细胞模型中进行几项基因操作 并依赖于核心中提供的分子生物学专业知识。原位研究的超微结构技术 通过神经解剖学家的参与，对动物模型的组织切片进行分析 在科罗拉多州立大学医疗中心担任Core的联合负责人(见分包合同)。这使所有调查人员能够 以非常高的分辨率检查正在研究的动物模型的实际组织。的具体目标 核心C是：(1)维护关系数据库，该数据库集成了所使用的动物的所有信息 他们计划并持续更新每种动物组织和细胞培养实验的数据 模型；(2)协调涉及动物的活动，包括收获组织、杂交和 对转基因小鼠进行基因分型，维持特殊饮食，监测寿命，并保持充分的记录 记录；(3)准备和维护所需的细胞系和原代肌肉和神经元培养 (4)协助研究人员设计和使用新的分子生物试剂，包括 细胞转染的性能和异常故障的排除；(5)为动物准备迎接光线和 电子显微镜研究并进行脑、肌肉和其他组织的超微结构分析 在项目中指定的。核心C级工作人员都具有丰富的技术经验， 并因此使调查人员能够迅速交流新的成果，并 进行远远超出每个实验室小组可用专业知识或设施的实验。

All projects in this Program make use of both animal models and cell culture systems to investigate the roles of Ca2+ dysregulation, oxidative stress, and chronic excitotoxicity in aging. The Animal Models, Electron Micrsocopy, Cell Culture, and Molecular Biology Core is a point of convergence for a diverse set of resources, services, and expertise not available in individual laboratories. The Core will maintain the web accessible relational database for the tracking each animal in the program and continuously updating data obtained with each of the animals and the cell culture experiments. This site integrates the outcomes of the diverse services the Core provides and greatly facilitates information sharing among all participants. The F344BNF1 rats from the NIA colony serve as the model for in vivo biological aging. In addition, a novel transgenic mouse over-expressing a glutamate-synthesizing enzyme, GLUD1, in neurons provides a model of excess glutamate release leading to degenerative changes in brain and attenuated longevity. The staff of Core C maintain the mouse colony, including the genotyping, cross breeding and monitoring of longevity. Cell culture models related to the animal systems are prepared and maintained in the Core to permit mechanistic testing of observations from studies of intact animals. Cell models include C2C12 myocytes, primary skeletal muscle cells, primary neurons from embryonic F344BNF1 rats and GLUD1/wt mice, and neuronal SH-SY5Y cell lines. Investigators will carry out several genetic manipulations in the cell models and rely on the molecular biological expertise provided in the Core. Ultrastructural expertise for the in situ analysis of tissue sections from the animal models is provided through the involvement of a neuroanatomist at the KU Medical Center as a co-leader for the Core (See subcontract). This enables all investigators to examine at very high resolution the actual tissues from the animal models under study. Specific aims of Core C are to: (1) maintain the relational database that integrates all information about the animals used in the projects and continuously updates data from experiments with each animal tissue and the cell culture models; (2) coordinate activities involving animals, including harvesting tissues, cross-breeding and genotyping transgenic mice, maintaining special diets, monitoring longevity, and maintaining fullydocumented records; (3) prepare and maintain cell lines and primary muscle and neuronal cultures needed within projects; (4) assist investigators in designing and using new molecular biological reagents, including the performance of cell transfections and troubleshooting anomalies; (5) prepare animals for light and electron microscopy studies and conduct Ultrastructural analyses of brain, muscle, and other tissues specified in the projects. Core C staff members are all experienced in the techniques required for their contributions and, as a result, enable the investigators to have rapid exchange of new results and to undertake experiments that go much beyond the expertise or facilities available in each lab group.