Benzyl alcohol differently affects fluid phase endocytosis and exocytosis in renal epithelial cells

The effects of benzyl alcohol, a local anaesthetic commonly used for modification of membrane fluidity, on fluid phase endocytosis and on exocytosis have been investigated in MDCK cells. Fluid phase endocytosis in confluent cells monolayer grown on solid support was determined, at 37 degrees C, by the uptake of the fluorescent dye Lucifer Yellow (LY). Exocytosis was estimated from the release of LY by cells preloaded with the dye. Addition of benzyl alcohol resulted in a concentration dependent inhibition of fluid phase endocytosis. For 30 mM benzyl alcohol, the inhibition obtained (83%) compared with that produced by preincubating the cells in a solution made hypertonic with 0.25 M sucrose. The inhibitory effect of benzyl alcohol was reversed within 30 min by washing. Endocytosis inhibition by benzyl alcohol was also observed in LLC-PK1 cells and OK cells, two renal epithelial cell lines of proximal tubule origin. In contrast, benzyl alcohol had no effect on exocytosis in LLC-PK1 cells, a limited but significant (15% at 30 mM) stimulatory effect on exocytosis in MDCK cells and a marked stimulatory effect (75% at 30 mM) in OK cells. These data demonstrate that benzyl alcohol affects endocytosis and exocytosis processes in renal epithelial cells. They suggest that membrane fluidity may alter membrane trafficking in living renal epithelial cells.