Low temperature blocks fluid-phase pinocytosis and receptor-mediated endocytosis in Trypanosoma cruzi epimastigotes

Abstract

Gold-labeled albumin and transferrin were used to follow at the ultrastructural level the early events and the effect of low temperature on protein uptake by Trypanosoma cruzi epimastigotes. In parasites incubated for 5 min at 28 °C with protein-gold complexes, extracellular markers were found only at the cytostome and/or the flagellar pocket regions, whereas intracellular gold particles were detected inside small uncoated vesicles located nearby. Within 10 min, labeling was also observed in uncoated vesicles close to the nucleus. Only after 30 min could the tracers be detected in the reservosomes. Weak labeling in the cytostome and flagellar pocket of parasites incubated at 4 °C with the albumin-gold solution indicated that albumin uptake occurred by fluid-phase pinocytosis. On the other hand, intense labeling at the cytostome was observed in parasites incubated at 4 °C with gold-labeled transferrin, showing that receptor-mediated endocytosis occurs mainly at this site. Both proteins were absent from the cells at 4 °C and 12 °C. Raising the temperature from 12 °C to 28 °C led to transferrin labeling in intracellular vesicles dispersed throughout the cytoplasm, but not in reservosomes. Our results suggest that low temperatures affect the transport and pinching of endocytic vesicles as well as the rate of delivery of transferrin to reservosomes.