Fluid-phase uptake by macropinocytosis in Dictyostelium

J Cell Sci. 1997 Jan:110 ( Pt 2):105-12. doi: 10.1242/jcs.110.2.105.

Abstract

To study fluid-phase endocytosis in living cells and its relationship to changes in the cell cortex, we have used a green fluorescent protein (GFP)-tagged version of coronin, an actin-associated protein that localises to dynamic regions of the Dictyostelium cell cortex. In the confocal microscope, internalisation of fluorescently labelled dextran as a fluid-phase marker can be recorded simultaneously with the recruitment of the coronin-GFP fusion-protein from the cytoplasm of the phagocyte. At crown-shaped surface protrusions, extracellular medium is taken up into vesicles with an average diameter of 1.6 microns, which is significantly larger than the 0.1 microns diameter of clathrin-coated pinosomes. The observed frequency of macropinosome formation can account for a large portion, if not all, of the fluid-phase uptake. The redistribution of coronin-GFP strongly resembles cytoskeletal rearrangements during phagocytosis. Scanning-electron micrographs indicate that crown-shaped cell-surface extensions can undergo shape changes, without a particle bound, that are similar to shape changes that occur during phagocytosis. In quantitative assays, the uptake of particles and fluid are about equally dependent on F-actin and coronin.

MeSH terms

  • Actins / metabolism*
  • Animals
  • Dictyostelium / physiology*
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / pharmacokinetics
  • Microfilament Proteins / genetics
  • Microfilament Proteins / pharmacokinetics*
  • Pinocytosis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / pharmacokinetics

Substances

  • Actins
  • Luminescent Proteins
  • Microfilament Proteins
  • Recombinant Fusion Proteins
  • coronin proteins
  • Green Fluorescent Proteins