Effect of the amino acid analog hadacidin on intracellular membrane flow and the cell surface in Dictyostelium discoideum: A transmission- and scanning-electron-microscope study

Abstract

We examined the effect of the amino acid analog hadacidin ( N-formyl- N-hydroxy glycine) on the process of endocytosis in the slime mold Dictyostelium discoideum. Endocytosis was followed using iron-dextran and transmission electron microscopy. In cells taken from the mid-log growth stage, iron-dextran was found to be distributed in small, medium, and large vesicles at a density lower than that present in the incubation medium, thus suggesting the fusion of small, iron-dextran-containing pinosomal vesicles with intracellular vesicles not containing iron-dextran. In cells treated with hadacidin, more small vesicles were present than in untreated cells, there being a reduction in the number of larger-sized vesicles; in these vesicles, iron-dextron was present at a density similar to that of the medium. This result is consistent with the conclusion that, while pinocytosis had continued, the fusion of vesicles and dilution of the vesicle contents had been inhibited. Also, the large number of small pinosomal vesicles in the drug-treated cells suggested that the recycling of vesicles to the surface had been inhibited. The observation that pinocytosis but not recycling continued after drug treatment raised the question of the origin of the membrane needed for the formation of pinosomes. Measurements of the cell surface revealed no difference between drug-treated and untreated cells, indicating that, when the membrane was internalized for pinosomes, the cell size remained constant. Scanning electron microscopy of the surface of hadacidin-treated cells revealed that the filopodia normally seen on their surface had disappeared, suggesting that the filopodial membrane may serve as a reservoir of the cellular membrane destined to replace the membrane utilized in pinosome formation.