Kinetics of fluid-phase pinocytosis in Dictyostelium discoideum amoebae

Abstract

Kinetics of pinocytosis in Dictyostelium discoideum were investigated over an extended period of time (up to 6 hours) using fluorescein isothiocyanate (FITC)-dextran as a fluid-phase marker. FITC-dextran added to the medium accumulated rapidly inside the cells with a rate of influx equivalent to 9 microns3 of fluid/cell x min. After a period of about 90 min of uptake, the intracellular FITC-dextran level reached a plateau which corresponded to a strict balance between pinocytosis and exocytosis as shown both by efflux measurements and pulse experiments with (3H) dextran. At equilibrium, the amount of internalized marker reached a value equivalent to 790 microns3 of fluid taken up per amoeba, i.e. a volume paradoxically higher than the total aqueous space of the cell (520 microns3 ). FITC-dextran was thus markedly concentrated intracellularly. The endocytic compartment in which the intracellular FITC-dextran was concentrated could be completely washed out when FITC-dextran was removed from the external medium.