Fate of plasma membrane during endocytosis. II. Evidence for recycling (shuttle) of plasma membrane constituents.
Cultured rat embryo fibroblasts were first allowed to store for 24 h fluorescein-labeled goat immunoglobulins directed against rabbit immunoglobulins (F anti-R IgG), and were subsequently exposed for 24 h to [(3)H]acetylated rabbit immunoglobulins known to bind to the cell membrane either specifically (anti-plasma membrane IgG: A anti-PM IgG) or unspecifically (contol IgG: AC IgG). As a result of immunological interaction between the two antibodies (no effect was found if the cells had been preloaded with control goat FC IgG), a substantial portion of the stored F anti-R IgG was unloaded from its intracellular storage site, appearing in the medium in the form of soluble immune complexes with rabbit A IgG. Part of the unloaded F anti-R IgG also was recovered in association with the plasma membrane, but only when A anti-PM IgG was used. In addition, significant reverse translocation of AC IgG from plasma membrane to lysosomes or some related intracellular storage compartment was also observed. With A anti-PM IgG, this translocation was less marked and affecte at the same time the plasma membrane marker 5'- nucleotidase. Cells that had stored horseradish peroxidase (HRP) simultaneously with F anti-R IgG did not unload HRP when exposed to A anti-PM IgG. These results support strongly, though not unequivocally, the concept that plasma membrane patches interiorized by endocytosis are recycled, or shuttled, back to the cell surface. In the framework of this concept, recycling antibody-coated membrane is taken to serve as vehicle for the selective intracellular capture and extracellular discharge of immunologically bound F anti-R IgG. The alternative explanation of regurgitation triggered off by immune complexes is considered less likely in view of the lack of HRP unloading.
D000818:Animals; D001666:Binding Sites, Antibody; D002462:Cell Membrane / Q000276:immunology / Q000502:physiology*; D002478:Cells, Cultured; D004705:Endocytosis; D005347:Fibroblasts; D006041:Goats / Q000276:immunology; D006735:Horseradish Peroxidase / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology; D008247:Lysosomes / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D051381:Rats
D007074:Immunoglobulin G; D006735:Horseradish Peroxidase