Receptor-mediated endocytosis and intracellular trafficking of lipoproteins and transferrin in insect cells.
While the intracellular pathways of ligands after receptor-mediated endocytosis have been studied extensively in mammalian cells, in insect cells these pathways are largely unknown. We transfected Drosophila Schneider line 2 (S2) cells with the human low-density lipoprotein (LDL) receptor (LDLR) and transferrin (Tf) receptor (TfR), and used endocytosis of LDL and Tf as markers. After endocytosis in mammalian cells, LDL is degraded in lysosomes, whereas Tf is recycled. Fluorescence microscopy analysis revealed that LDL and Tf are internalized by S2 cells transfected with LDLR or TfR, respectively. In transfectants simultaneously expressing LDLR and TfR, both ligands colocalize in endosomes immediately after endocytic uptake, and their location remained unchanged after a chase. Similar results were obtained with Spodoptera frugiperda Sf9 cells that were transfected with TfR, suggesting that Tf is retained intracellularly by both cell lines. The insect lipoprotein, lipophorin, is recycled upon lipophorin receptor (LpR)-mediated endocytosis by mammalian cells, however, not after endocytosis by LpR-expressing S2 transfectants, suggesting that this recycling mechanism is cell-type specific. LpR is endogenously expressed by fat body tissue of Locusta migratoria for a limited period after an ecdysis. A chase following endocytosis of labeled lipophorin by isolated fat body tissue at this developmental stage resulted in a significant decrease of lipophorin-containing vesicles, indicative of recycling of the ligand.
Van Hoof
Dennis
D
Department of Biochemical Physiology and Institute of Biomembranes, Utrecht University, Padualaan 8, Room W-209, Utrecht 3584 CH, The Netherlands.
Rodenburg
Kees W
KW
Van der Horst
Dick J
DJ
D000818:Animals; D002460:Cell Line; D004330:Drosophila / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D005216:Fat Body / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007313:Insecta / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism*; D021381:Protein Transport; D011973:Receptors, LDL / Q000235:genetics / Q000502:physiology*; D011990:Receptors, Transferrin / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism*
D008074:Lipoproteins; D011973:Receptors, LDL; D011990:Receptors, Transferrin; D011994:Recombinant Proteins; D014168:Transferrin