All projects in this Program make use of both animal models and cell culture systems to investigate the rolesof Ca2+ dysregulation, oxidative stress, and chronic excitotoxicity in aging. The Animal Models, ElectronMicrsocopy, Cell Culture, and Molecular Biology Core is a point of convergence for a diverse set ofresources, services, and expertise not available in individual laboratories. The Core will maintain the webaccessible relational database for the tracking each animal in the program and continuously updating dataobtained with each of the animals and the cell culture experiments. This site integrates the outcomes of thediverse services the Core provides and greatly facilitates information sharing among all participants. TheF344BNF1 rats from the NIA colony serve as the model for in vivo biological aging. In addition, a noveltransgenic mouse over-expressing a glutamate-synthesizing enzyme, GLUD1, in neurons provides a modelof excess glutamate release leading to degenerative changes in brain and attenuated longevity. The staff ofCore 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 permitmechanistic 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, andneuronal SH-SY5Y cell lines. Investigators will carry out several genetic manipulations in the cell modelsand rely on the molecular biological expertise provided in the Core. Ultrastructural expertise for the in situanalysis of tissue sections from the animal models is provided through the involvement of a neuroanatomistat the KU Medical Center as a co-leader for the Core (See subcontract). This enables all investigators toexamine at very high resolution the actual tissues from the animal models under study.
Specific aims ofCore C are to: (1) maintain the relational database that integrates all information about the animals used inthe projects and continuously updates data from experiments with each animal tissue and the cell culturemodels; (2) coordinate activities involving animals, including harvesting tissues, cross-breeding andgenotyping transgenic mice, maintaining special diets, monitoring longevity, and maintaining fullydocumentedrecords; (3) prepare and maintain cell lines and primary muscle and neuronal cultures neededwithin projects; (4) assist investigators in designing and using new molecular biological reagents, includingthe performance of cell transfections and troubleshooting anomalies; (5) prepare animals for light andelectron microscopy studies and conduct Ultrastructural analyses of brain, muscle, and other tissuesspecified in the projects. Core C staff members are all experienced in the techniques required for theircontributions and, as a result, enable the investigators to have rapid exchange of new results and toundertake experiments that go much beyond the expertise or facilities available in each lab group.
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