The long-term objectives of this proposal are to discover fundamental new knowledge about the proliferative capacity of human neurons, to apply this knowledge to age- related changes in the brain, and to discover drugs for the treatment of neurological injuries and disorders. The focus of the latter will be on stroke, myocardial infarction and traumatic head injury models in which ischemic damage leads to the opening of calcium channels and excessive release of glutamate, causing cell damage and, ultimately, cell death. Thus, effective inhibitors of human glutamate receptors and calcium channels are of clinical importance. To attain these goals, human neuronal lines from the central nervous system (CNS) will be developed.From these we will gain information about the ability of human neurons to retain their phenotype after proliferation, and they will provide an assay system for screening glutamate and calcium channel antagonists that may be clinically useful. Currently, only few human neuronal lines exist. In the past, most studies have been carried out with rodent cells; the results were then extrapolated to human, with the caveat that differences between species may exist. Therefore, the establishment of human cell lines from specific regions of the central nervous system would offer distinct advantages for testing neuropharmacological agents for clinical applications. The specific aims of this project are: to optimize conditions for growing perpetualized and immortalized human neurons from the CNS that retain neuronal phenotype, and to characterize their glutamate receptors and calcium channels, so that they may eventually be used to screen drug candidates.
