Cell and Tissue Research

, Volume 320, Issue 1, pp 99–113

Transferrin recycling and dextran transport to lysosomes is differentially affected by bafilomycin, nocodazole, and low temperature

Authors

  • Günther Baravalle
    • Department of Pathophysiology, Center for Physiology and PathophysiologyMedical University of Vienna
  • Daniela Schober
    • Department of Pathophysiology, Center for Physiology and PathophysiologyMedical University of Vienna
  • Marlis Huber
    • Department of Pathophysiology, Center for Physiology and PathophysiologyMedical University of Vienna
  • Nora Bayer
    • Department of Pathophysiology, Center for Physiology and PathophysiologyMedical University of Vienna
    • PerkinElmer Life and Analytical Sciences Vienna
  • Robert F. Murphy
    • Departments of Biological Sciences and Biomedical EngineeringCarnegie Mellon University
    • Department of Pathophysiology, Center for Physiology and PathophysiologyMedical University of Vienna
Regular Article

DOI: 10.1007/s00441-004-1060-x

Cite this article as:
Baravalle, G., Schober, D., Huber, M. et al. Cell Tissue Res (2005) 320: 99. doi:10.1007/s00441-004-1060-x

Abstract

The effects of bafilomycin, nocodazole, and reduced temperature on recycling and the lysosomal pathway have been investigated in various cultured cell lines and have been shown to vary dependent on the cell type examined. However, the way in which these treatments affect recycling and transport to lysosomes within the same cell line has not been analyzed. In the current study, we used fluorophore-labeled transferrin and dextran as typical markers for the recycling and the lysosomal pathways, respectively, to explore the morphology and the intravesicular pH of endocytic compartments in HeLa cells. The V-ATPase inhibitor bafilomycin selectively inhibited the transport of marker destined for lysosomal degradation in early endosomes, whereas the transport of transferrin to the perinuclear recycling compartment (PNRC) still occurred. The kinetics of transferrin acidification was found to be biphasic, indicative of fast and slow recycling pathways via early endosomes (pH 6.0) and PNRC (pH 5.6), respectively. Furthermore, the disruption of microtubules by nocodazole blocked the transport of transferrin to the PNRC in early endosomes and of lysosome-directed marker into endosomal carrier vesicles. In contrast, incubation at 20°C affected the lysosomal pathway by causing retention of internalized dextran in late endosomes and a delay in transferrin recycling. Taken together, these data clearly demonstrate, for the first time, that the transferrin recycling pathway and transport of endocytosed material to lysosomes are differentially affected by bafilomycin, nocodazole, and low temperature in HeLa cells. Consequently, these treatments can be applied to investigate whether internalized macromolecules such as viruses follow a recycling or degradative pathway.

Keywords

Endocytic pathwaysBafilomycinNocodazoleLow temperatureEndosomal pHHeLa Ohio cells (Wisconsin strain)

Copyright information

© Springer-Verlag 2005