The fine topological relationship between sinus-lining endothelial cells (SLE) and vessel-lining endothelial cells (VLE) at the opening portion of sinusoids into central or interlobular veins of rat liver was studied by a comparison of morphological and functional properties of both types of cells. Three minutes after intravenous injection of formalin-denatured albumin conjugated with horseradish peroxidase (HRP-FDA), liver was perfused with fixative. Chopped sections of the liver (50 micron thick) were incubated in diaminobenzidine-H2O2 medium, followed by processing for electron microscopy. The HRP-FDA was localized in endocytotic vesicles and vacuoles of the SLE and Kupffer cells but not of the VLE lining interlobular or central veins or interlobular arteries. In the opening portion of the sinusoids into these veins, the attenuated cytoplasmic extensions of the SLE containing positive vesicles were in direct contact with squamous process of the VLE having no positive vesicles. The contact was mediated by overlapping junctions. No intermediate cell type between the SLE and VLE in this region or other portions was noted. The results indicate that the habitat of the SLE is exactly isolated from that of the VLE in rat liver and at the transitional portion from sinusoids to veins or arteries they are directly connected with each other by overlapping junctions.