Receptor-mediated endocytosis (RME) by a scavenger receptor on sinusoidal liver endothelial cells (LECs) for formaldehyde-treated bovine serum albumin (f-Alb) has previously been shown to be impaired following chronic ethanol consumption. These studies were initially performed by in situ perfusion, making it difficult to determine the point in the process at which RME is affected. Therefore, it was the purpose of this study to use isolated LECs to begin elucidating at what point in the process chronic ethanol consumption affects RME. Initial studies showed that degradation at the single-cell level were similarly decreased at levels that had been observed for in situ studies, suggesting that the ethanol effects can be repeated using isolated LECs, making them useful for in vitro studies. Binding studies with 125I-formaldehyde-treated bovine serum albumin (125I-f-Alb) demonstrated there was a slight, but significantly different, decrease in binding by LECs from ethanol-fed rats when compared with pair-fed or chow-fed rats. However, the affinity of these receptors was not different between these groups. In contrast, a defect in the initial stages of receptor-ligand internalization was indicated as less surface-bound ligand was internalized and subsequently degraded in cells from the ethanol-treated animals as compared with controls. Additionally, once the data were adjusted for the amount of ligand internalized, the degradation of the internalized ligand was only slightly impaired. These results indicate that chronic ethanol feeding impairs the process of RME by the liver; the major cause of this impairment appears to be caused by a decreased ability of these cells to internalize all of the surface-bound ligand, with a minimal defect in postinternalization events.