Endocytic pathway of scavenger receptors via trans-Golgi system in bovine alveolar macrophages

Abstract

Background: The scavenger receptors are trimeric membrane glycoproteins present on the surface membrane of macrophages and macrophage-derived foam cells and are implicated in the deposition of cholesterol in the arterial walls during atherogenesis. This receptor mediates endocytosis of a diverse alley of negatively charged macromolecules including modified low density lipoproteins.

Experimental design: We tried biochemical and morphologic approaches to elucidate in more detail the endocytic and intracellular pathway of the scavenger receptors during endocytosis of acetylated low density lipoproteins (AcLDL) and oxidized LDL (OxLDL) by bovine alveolar macrophages. Biochemically, cell interaction and subsequent intracellular degradation of AcLDL and OxLDL were examined using [125I]AcLDL or [125I]OxLDL. At the ultramorphologic level, the scavenger receptors were detected by a mouse monoclonal antibody against bovine macrophages, D2, with colloidal gold particles (AcLDL-Au or OxLDL-Au particles) as tracer of ligands.

Results: The biochemical studies demonstrated that after binding of AcLDL and OxLDL to the cell surface, these ligands were internalized and degraded intracellularly. Immunoelectron microscopy revealed that after a 2-hour incubation at 4 degrees C AcLDL-Au or OxLDL-Au particles were bound to the scavenger receptors present on the cell surfaces. Upon a temperature shift from 4 to 37 degrees C, these particles were concentrated in coated pits and internalized in endosomes. After subsequent dissociation of the particles from the receptors within the endosomes, the ligands were transported to secondary lysosomes, the receptors were transported to a trans-Golgi system. In the Golgi complex, the receptors were packed in secretory vesicles and recycled to the cell surface membrane. The transport and recycling of the receptors via the trans-Golgi system were completely inhibited by incubation below 18 degrees C or pretreatment with monensin.

Conclusions: These results suggest that during endocytosis of modified low density lipoprotein, the Golgi complex plays a crucial role in sorting and recycling of the scavenger receptors in macrophages.