Hepatic uptake and intracellular processing of LDL in rainbow trout

The process of receptor-mediated endocytosis in poikilothermic vertebrates such as salmonid fish have not been subjected to much research, compared to the detailed studies done in mammalian systems. We have investigated the hepatic uptake and intracellular processing of low-density lipoprotein (LDL) in rainbow trout liver. After intravenous injection of the [125I]tyramine-cellobiose ([125I]TC) -labelled lipoprotein, the liver was perfused and cells isolated or fractionated by differential centrifugation and isopycnic centrifugation in Nycodenz gradients. We found that LDL was mainly endocytosed by parenchymal cells of the liver. Cell fractionation experiments showed that LDL was localized sequentially in three groups of organelles of increasing density. Initially, LDL was localized in small, slowly sedimenting endosomes before being transferred to denser endosomes (prelysosomes) and finally to dense lysosomes. The lysosomes were identified by three lysosomal marker enzymes. Degradation products formed from [125I]TC-labelled LDL could also be detected in prelysosomal vesicles. In vitro experiments with cultured trout hepatocytes demonstrated that intracellular processing of [125I]TC-LDL in these cells could be suppressed by endocytic and lysosomal inhibitors. The catabolism of LDL in rainbow trout therefore follows the endocytic-lysosomal pathway described for many macromolecules in mammalian cells.