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Endosomal density shift is related to a decrease in fusion capacity.

R Diaz1, M I Colombo, M Koval

  • 1Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis 63110.

European Journal of Cell Biology
|December 1, 1991
PubMed
Summary
This summary is machine-generated.

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Cellular probes reveal that endocytic vesicle density increases during lysosome targeting, impacting fusion activity. This density shift is a key step, distinct from fusion itself, in directing vesicles to lysosomes.

Area of Science:

  • Cell Biology
  • Endocytosis and Vesicular Transport
  • Lysosome Biogenesis

Background:

  • Dinitrophenol (DNP)-beta-glucuronidase and mannosylated anti-DNP IgG were previously established as cell-free probes for early endosome fusion.
  • These probes are endocytosed via the mannose receptor and trafficked to lysosomes.

Purpose of the Study:

  • To investigate the transport dynamics and fusion capabilities of endocytic probes in intact cells.
  • To elucidate the relationship between endocytic vesicle density and fusion activity in targeting to lysosomes.

Main Methods:

  • Utilized Percoll gradient fractionation to analyze the buoyant density of endocytic vesicles in cell homogenates.
  • Employed pulse-chase experiments with DNP-derivatized transferrin-alkaline phosphatase conjugate to track recycling ligands.

Related Experiment Videos

  • Investigated the role of microtubules by treating cells with nocodazole or colchicine.

    • Main Results:

    • In intact cells, probes were transported to endocytic vesicles of increasing buoyant density, correlating with decreased in vitro fusion activity.
    • Monensin treatment retained probes in lighter, fusion-competent vesicles, while recycling ligands consistently localized to light vesicles.
    • Ligand mixing occurred in both early and late endocytic compartments in intact cells, with microtubule disruption reducing mixing in late endosomes.

    Conclusions:

    • The observed increase in endocytic vesicle density is an initial step in lysosomal targeting, preceding or independent of fusion.
    • Fusion activity in vitro does not inherently increase vesicle density; the density shift in intact cells suggests a distinct maturation process.
    • Microtubules play a role in efficient ligand mixing within late endocytic compartments.