The overall objective of the proposed project is to evaluate the molecular mechanisms of hepatic protein synthesis, focusing on the process of cirrhosis, through the use of molecular hybridization technology. This will facilitate my long-term objective to become an independent investigator in academic hepatology with expertise in the current areas of molecular biology. Our use of much of this technology in our previous investigations of the effect of chronic ethanol exposure and uremia on albumin mRNA content indicates the feasability of the project. In addition, I will fuether develop my technological skills to analyze the basis for differential expression of the collagen gene. The specific areas of investigation include: 1) the evaluation of mechanisms involved in the increased synthesis of collagen in 2 animal models, murine schistosomiasis and CC14-induced cirrhosis in rats, using in vitro protein synthesis and molecular hybridization. We are expecially interested in evaluating the qualitative and quantitative changes in procollagen mRNA content in the cirrhotic animal. (2) the evaluation by similar technology of the effect of chronic ethanol exposure on the procollagen mRNA content in the cirrhotic liver (3) the identification by means of in situ hybridization of the specific cell type(s) that synthesizes collagen in the normal and fibrotic liver, and (4) the investigation of whether any human collagen gene polymorphism exists which might determine which chronic alcohol abusers will develop cirrhosis. This would include Southern blot hybridization of enzyme-restricted human genomic DNA, in vitro transcription assays and the use of specialized recombinant plasmid vectors. By accomplishing the aims of the proposal, we would hope to formulate a general model for the pathogenesis of hepatic fibrosis at the molecular level. Identification of human collagen gene polymorphism will serve as a clinically relevant means of predicting which alcohol abusers will develop cirrhosis. The environment provided by the Liver Research Center and the Departments of Cell Biology and Pathology, under the tutelage of Dr. d. Shafritz and others will allow me the develop my skills to the fullest and help train me in new areas of expertise.