In the present work we have studied the effect of prenatal exposure to alcohol on the synthesis, glycosylation, and transport of proteins in fetal hepatocytes isolated from 21-day-old fetuses derived from control and chronic alcoholic rats. Protein synthesis was evaluated both in a cell-free system and in hepatocytes after (35S)methionine and (3H)leucine incorporation, respectively. Glycosylation was assessed using (3H)mannose and (3H)galactose as precursors. Protein synthesis was significantly decreased in treated hepatocytes. In control hepatocytes, quantitative electron microscope autoradiography showed that both (3H)leucine and (3H)mannose incorporation occur first in the rough endoplasmic reticulum (rER). Later the silver grains appeared over the Golgi apparatus, and, finally there was a transport towards the cell periphery. After pulse, silver grains corresponding to (3H)galactose incorporation appeared over the Golgi apparatus. The label then moved to the hepatocyte periphery. Alcohol treated hepatocytes showed a retention of grains over the Golgi apparatus with a diminution in the label at the cell periphery. These results indicate that prenatal exposure to alcohol induces a decrease in the synthesis of proteins in the hepatocyte as well as an alteration in the process of glycosylation and/or transport of secretory proteins.