Clathrin-mediated endocytosis of high density lipoprotein, in human intestinal Caco-2 cells. A post-embedding immunocytochemical study

The mechanism by which high density lipoprotein (HDL) removes excess cholesterol from intracellular sites has been the subject of much controversy. There is some evidence that HDL binds to specific cell surface receptors without internalization. Other evidence suggests that HDL is taken up by endocytosis, enters a pathway of endosomal trafficking and is resecreted from the cells (retroendocytsosis). In the present study, we investigated the distribution of apolipoprotein AI, the major protein constituent of HDL, in cultured intestinal Caco-2 cells employing post-embedding immunocytochemistry on LR White-embedded material. Cells grown under control conditions showed label for apolipoprotein AI in the endoplasmic reticulum. After incubation with native apolipoprotein E-free high density lipoprotein, (HDL,) additional label for apolipoprotein AI was found in endosomes. These endosomes were observed near lipid droplets and in the basolateral cytoplasm. Further, it was demonstrated that label for apolipoprotein AI was cofocalized with label for clathrin on the basolateral membrane. Our results support the concept that HDL, is internalized and subsequently processed in an endosomal pathway in Caco-2 cells besides de novo synthesis of apolipoprotein AI.