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Vol. 10, Issue 12, 4419-4427, December 1999

Circulation of the Plasma Membrane in Dictyostelium

Carmen Aguado-Velasco,^* and Mark S. Bretscher

Medical Research Council Laboratory for Molecular Biology, Cambridge CB2 2QH, United Kingdom

We have developed a fluorimetric assay with the use of the dye FM1-43 to determine the rate at which Dictyostelium amoebae endocytose their surface membrane. Our results show that they do so about once each 4-10 min. A clathrin null mutant takes its surface up only ~30% more slowly, showing that this membrane uptake cannot be caused by clathrin-coated vesicles. Surprisingly, Ax2 and its parent, NC4, which differ in their rates of fluid-phase internalization by ~60-fold, take up their surfaces at the same rates. These results show that, in axenic cells, the uptake of fluid and of surface area are separate processes. The large activity of this new endocytic cycle in both Ax2 and NC4 amoebae appears capable of delivering sufficient new surface area to advance the cells' fronts during migration.

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^* Present address: Departamento de Anatomía, Universidad de Navarra, Pamplona, Spain.

Corresponding author. E-mail address: msb{at}mrc-lmb.cam.ac.uk.

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Molecular Biology of the Cell
Vol. 10, 4419-4427, December 1999
Copyright © 1999 by The American Society for Cell Biology

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