Independent variation in the number of coated pits and of coated vesicles in cultured fibroblasts.
When tissue culture cells were maintained at 37 degrees C in a serum-free medium for 4 hr no change in the number of coated pits could be detected using ultrastructural techniques. However, the number of coated vesicles was highly significantly increased, being 179% more than in the control cultures. If the cells were put back into a medium supplemented with 5% calf serum, the number of coated pits was unchanged, but the number of coated vesicles decreased and returned to the control level within a few minutes. The same results were obtained when using ligands such as Low Density Lipoprotein or alpha-2-macroglobulin which are known to be internalized via coated structures. It is concluded that coated pits appear and disappear at equal rates and that coated vesicles can accumulate independently. It is suggested that this could be due to the presence of a large reserve of soluble clathrin. This pool would have a low turnover rate because cycloheximide did not block coated vesicle accumulation over the period studied.
D000818:Animals; D001386:Azides / Q000494:pharmacology; D002478:Cells, Cultured; D003034:Coated Pits, Cell-Membrane / Q000187:drug effects / Q000648:ultrastructure*; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D004705:Endocytosis; D011992:Endosomes / Q000648:ultrastructure*; D005347:Fibroblasts / Q000648:ultrastructure*; D005476:Fluphenazine / Q000494:pharmacology; D000882:Haplorhini; D008024:Ligands
D001386:Azides; D003470:Culture Media; D008024:Ligands; D003513:Cycloheximide; D005476:Fluphenazine