ANIMAL MODELS, ELECTRON MICROSCOPY, CELL CULTURE AND MOLECULAR BIOLOGY

动物模型、电子显微镜、细胞培养和分子生物学

• 批准号: 8037701
• 负责人: MARY L. MICHAELIS
• 金额: $ 39.53万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8037701
• 关键词: Age; Aging; Aging-Related Process; Animal Model; Animals; Attenuated; Biological; Biological Aging; Brain; Breeding; Cell Culture System; Cell Culture Techniques; Cell Line; Cell model; Cells; Chronic; Data; Databases; Development; Diet Monitoring; Electron Microscopy; Electrons; Embryo; Enzymes; Event; Gene Targeting; Genes; Genotype; Glutamate Dehydrogenase; Glutamates; Goals; Hippocampus (Brain); Human; Hybrids; In Situ; Individual; Laboratories; Lead; Light; Longevity; Medical center; Membrane Microdomains; Metabolism; Modeling; Molecular; Molecular Biology; Monitor; Mus; Muscle; Muscle Cells; Muscle Fibers; Mutation; Neuroblastoma; Neurons; Norway; Outcome; Oxidative Stress; Participant; Pattern; Performance; Perfusion; Preparation; Protocols documentation; Rattus; Reagent; Records; Research; Research Personnel; Resolution; Resources; Role; Services; Signal Transduction; Site; Skeletal Muscle; Source; Specific qualifier value; Study models; System; Systems Integration; Techniques; Testing; Tissue Harvesting; Tissues; Training; Transfection; Transgenic Mice; Transgenic Organisms; Update; age related; aging brain; animal tissue; brain tissue; data sharing; design; excitotoxicity; experience; genetic manipulation; in vivo; in vivo Model; innovation; insight; member; novel; overexpression; programs; relational database; research study; sample fixation; success; tissue/cell culture; trend; virtual; web-accessible

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.

该计划中的所有项目均利用动物模型和细胞培养系统来研究其作用 衰老过程中的 Ca2+ 失调、氧化应激和慢性兴奋性毒性。动物模型，电子 显微镜、细胞培养和分子生物学核心是多种学科的汇聚点 个别实验室不具备的资源、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只动物并不断更新数据 通过每只动物和细胞培养实验获得。该网站整合了 Core提供的服务多样化，极大地促进了所有参与者之间的信息共享。这 来自 NIA 群体的 F344BNF1 大鼠作为体内生物衰老模型。另外，还有一本小说 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 过量的谷氨酸释放导致大脑退行性变化和寿命缩短。的工作人员 Core C 维护小鼠群体，包括基因分型、杂交育种和寿命监测。 与动物系统相关的细胞培养模型在核心中准备和维护，以允许 对完整动物研究的观察结果进行机械测试。细胞模型包括C2C12肌细胞， 原代骨骼肌细胞、来自胚胎 F344BNF1 大鼠和 GLUD1/wt 小鼠的原代神经元，以及 神经元 SH-SY5Y 细胞系。研究人员将在细胞模型中进行多项基因操作 并依赖核心中提供的分子生物学专业知识。原位超微结构专业知识 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 KU 医疗中心担任核心联合领导者（参见分包合同）。这使得所有调查人员能够 以非常高分辨率检查所研究的动物模型的实际组织。具体目标 核心 C 是：（1）维护关系数据库，该数据库集成了有关所用动物的所有信息 项目并不断更新每种动物组织和细胞培养物的实验数据 模型； (2) 协调涉及动物的活动，包括收获组织、杂交育种和 对转基因小鼠进行基因分型、维持特殊饮食、监测寿命并维持完整记录 记录； (3) 准备和维持所需的细胞系和原代肌肉和神经元培养物 在项目内； （4）协助研究者设计和使用新型分子生物学试剂，包括 细胞转染的表现和异常故障排除； (5) 准备动物光照和 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 项目中指定。核心 C 人员在其所需的技术方面都拥有丰富的经验 贡献，从而使研究人员能够快速交流新结果并 进行远远超出每个实验室组现有专业知识或设施的实验。