Reversible plasma membrane ultrastructural changes correlated with electropermeabilization in Chinese hamster ovary cells

Abstract Chinese hamster ovary cells (CHO) grown in monolayers were permeabilized to molecules with molecular weight up to 1000 by high intensity 100 μs square wave electric field pulses. This permeability was transient and the cell viability was not affected. It was not possible for molecules with a molecular weight larger than 1500 to penetrate inside the cytoplasm if lytic pulsing conditions were not used. In order to investigate the ultrastructural changes associated with this transient and limited permeabilization, cells were chemically fixed a few seconds after their pulsation and observed by electron microscopy. By scanning electron microscopy, numerous microvilli and blebs were observed almost immediately after application of the field. No other membrane changes were observed. Permeabilization of the membrane was visualized at the electron microscopic level by penetration of Ruthenium red. The appearance of osmotic pressure-dependent ‘blebs’ was indicative of local weakening of the plasma membrane. Most of these effects were fully reversible and disappeared within 30 min at 37 °C with the formation of huge polykaryons when cells were in contact before pulsing.