(from the application): A multi-disciplinary approach is proposed in this Program Project to define and characterize the signal transduction pathways by which insulin and the gonadal hormones, estrogen and testosterone, regulate the metabolic fate of the Alzheimer precursor protein (APP) and influence beta-amyloid peptide (ABeta) formation and deposition. 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 (AD). A broad range of studies will be performed at distinct levels of organizational complexity to define the effects of the hormonal regulation of APP/ABeta metabolism, encompassing in vitro biochemical studies with purified enzymes and other molecules cellular and cell-free systems that accommodate experimental manipulation, and studies in intact animals analyzing both biochemical and morphological changes in APP/ABeta metabolism that can be observed as consequence of the experimental depletion and replacement of the hormones. Scientific Core is designed to provide a reliable source for the production and supply of animals cultured neurons, antibodies and other key reagents and the performance of routine, yet essential tasks that will be required to accomplish the studies described in the research Projects. The centralized organization of support functions will facilitate an efficient and cost-effective means to ensure an adequate supply of materials. The Core staff will maintain the colonies of transgenic AD mice and perform genotyping of offspring (Specific Aim 1), maintain stocks of reagents in current use (Specific Aim 2), and will be responsible for the design screening, and purification of new polyclonal antibodies that are required, including epitope-, domain-, and phosphorylation state-specific antibodies to APP/ABeta fragments and? the cdk5/p35/p25 enzyme complex (Specific Aim 3). The Core will maintain a tissue culture facility to provide a continuous supply of primary cultures of rat and mouse embryonic neurons to all three Projects (Specific Aim 4).
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