Hepatic lipase is localized at the parenchymal cell microvilli in rat liver.
Hepatic lipase (HL) is thought to be located at the vascular endothelium in the liver. However, it has also been implicated in the binding and internalization of chylomicron remnants in the parenchymal cells. In view of this apparent discrepancy between localization and function, we re-investigated the localization of HL in rat liver using biochemical and immunohistochemical techniques. The binding of HL to endothelial cells was studied in primary cultures of rat liver endothelial cells. Endothelial cells bound HL in a saturable manner with high affinity. However, the binding capacity accounted for at most 1% of the total HL activity present in the whole liver. These results contrasted with earlier studies, in which non-parenchymal cell (NPC) preparations had been found to bind HL with a high capacity. To study HL binding to the different components of the NPC preparations, we separated endothelial cells, Kupffer cells and blebs by counterflow elutriation. Kupffer cells and endothelial cells showed a relatively low HL-binding capacity. In contrast, the blebs, representing parenchymal-cell-derived material, had a high HL-binding capacity (33 m-units/mg of protein) and accounted for more than 80% of the total HL binding in the NPC preparation. In contrast with endothelial and Kupffer cells, the HL-binding capacity of parenchymal cells could account for almost all the HL activity found in the whole liver. These data strongly suggest that HL binding occurs at parenchymal liver cells. To confirm this conclusion in situ, we studied HL localization by immunocytochemical techniques. Using immunofluorescence, we confirmed the sinusoidal localization of HL. Immunoelectron microscopy demonstrated that virtually all HL was located at the microvilli of parenchymal liver cells, with a minor amount at the endothelium. We conclude that, in rat liver, HL is localized at the microvilli of parenchymal cells.
Breedveld
B
B
Department of Biochemistry, Erasmus University Rotterdam, The Netherlands.
Schoonderwoerd
K
K
Verhoeven
A J
AJ
Willemsen
R
R
Jansen
H
H
D000818:Animals; D002478:Cells, Cultured; D004727:Endothelium / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D007150:Immunohistochemistry; D008049:Lipase / Q000378:metabolism* / Q000648:ultrastructure; D008099:Liver / Q000166:cytology / Q000201:enzymology* / Q000648:ultrastructure*; D008297:Male; D016253:Microscopy, Immunoelectron; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar
D008049:Lipase