The physiological regulation of macropinocytosis during Dictyostelium growth and development

Abstract

Macropinocytosis is a conserved endocytic process used by Dictyostelium amoebae for feeding on liquid medium. To further Dictyostelium as a model for macropinocytosis, we developed a high-throughput flow cytometry assay for macropinocytosis, and used it to identify inhibitors and investigate the physiological regulation of macropinocytosis. Dictyostelium has two feeding states: phagocytic and macropinocytic. When cells are switched from phagocytic growth on bacteria to liquid media, the rate of macropinocytosis slowly increases, due to increased size and frequency of macropinosomes. Upregulation is triggered by a minimal medium of 3 amino acids plus glucose and likely depends on macropinocytosis itself. Bacteria suppress macropinocytosis while their product, folate, partially suppresses upregulation of macropinocytosis. Starvation, which initiates development, does not of itself suppress macropinocytosis: this can continue in isolated cells, but is shut down by a conditioned-medium factor or activation of PKA signalling. Thus macropinocytosis is a facultative ability of Dictyostelium cells, regulated by environmental conditions that are identified here.\n\nSummaryA high-throughput flow cytometry assay shows that macropinocytosis in D. discoideum is upregulated in the presence of nutrients and absence of bacteria. Development and bacteria induce cells to downregulate macropinocytosis.