ANIMAL MODELS, ELECTRON MICROSCOPY, CELL CULTURE AND MOLECULAR BIOLOGY

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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.