A variety of virus vectors have established that effective gene delivery can be attained in non-dividing mammalian cells. For example, adeno- associated virus (AAV) vectors have been shown to stably transfer and express foreign genes in brain and muscle with little or no accompanying toxicity (McCown et al., 1996; Xiac et al., 1996). Based upon these advances, the present center grant will focus upon the use of AAV and adenovirus (AD) vectors to deliver and express genes proposed to ameliorate the adverse effects of chronic ethanol exposure, both in brain and liver. Therefore, the role of the vector core will be able to construct the cDNA cassettes, insert these cassettes into AAV of Ad vector plasmids, replicate and package the virus vectors, and finally test the function of the vectors, both in vitro and in vivo. Specifically, in Dr. Crews' section, AAV-tyrosine hydroxylase or tryptophan hydroxylase vectors will be prepared to test the involvement of dopaminergic and/or serotonergic function in rodent models of ethanol preference. In Dr. Thurman's section, AAV vectors will be prepared for the delivery of superoxide dismutase (SOD)/catalase, in order to evaluate the origin of ethanol induced oxidative damage to the liver, while Dr. Sulik will use the same AAV vectors to probe the mechanisms of ethanol teratogenicity. For Dr. Brenner, AD vectors will be prepared in order to identify the role of the immediate early genes, APl and NFkappaB, in ethanol-induced ethanol liver pathology. For Dr. Murrow, both sense and antisense AAV vectors will be prepared to delineate the role of GABAa receptor subunits in ethanol self administration, and finally, AAV vectors with antisense constructs to diazepam-binding inhibitor (DBI) and corticotrophan-releasing hormone (CRF) will be prepared for Dr. Breese to probe the role of these factors in the evolution of ethanol withdrawal anxiety. Thus, the vector core will provide the means to investigate molecular mechanisms directly involved in ethanol-induced pathologies, both in the brain and in the liver.
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