Inhibition of pinocytosis in rat embryo fibroblasts treated with monensin.
Rat embryo fibroblasts cultured in the presence of monensin exhibited an inhibited uptake of horseradish peroxidase. The inhibition was detected after 3 h, after which time the cells became increasingly vacuolated; the concentration of monensin required to inhibit pinocytosis (0.4 microM for half-maximum inhibition at 18 h) was similar to that found by others to inhibit secretion. Both the exchange of 5'-nucleotidase between the membranes of cytoplasmic organelles and the cell surface and the internalization of anti-5'-nucleotidase bound to the cell surface were inhibited by approximately 90% in monensin-treated cells. The effects of monensin were reversible: cells cultured first with monensin, and then in fresh medium, exhibited control levels of horseradish peroxidase uptake, exchange of 5'-nucleotidase, and internalization of anti-5'-nucleotidase bound to the cell surface. After monensin treatment, the median density of both galactosyl transferase and 5'-nucleotidase increased from 1.128 to 1.148, and the median density of both N-acetyl-beta-glucosaminidase and horseradish peroxidase taken up by endocytosis decreased from 1.194 to 1.160. The results indicate that monensin is a reversible inhibitor of pinocytosis and, presumably, therefore, of membrane recycling. They suggest that the inhibition of membrane recycling occurs at a step other than the fusion of pinocytic vesicles with lysosomes and is perhaps a consequence of an effect of the ionophore on the Golgi complex.
D015720:5'-Nucleotidase; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005347:Fibroblasts; D005663:Furans / Q000494:pharmacology*; D006735:Horseradish Peroxidase; D007425:Intracellular Membranes / Q000201:enzymology; D008247:Lysosomes / Q000187:drug effects; D008985:Monensin / Q000494:pharmacology*; D009708:Nucleotidases / Q000378:metabolism; D010873:Pinocytosis / Q000187:drug effects*; D014176:Protein Biosynthesis; D051381:Rats; D014617:Vacuoles / Q000648:ultrastructure
D005663:Furans; D000255:Adenosine Triphosphate; D008985:Monensin; D006735:Horseradish Peroxidase; D009708:Nucleotidases; D015720:5'-Nucleotidase