Membrane disorder and phospholipid scrambling in electropermeabilized and viable cells

• Published in: Biochimica et biophysica acta Vol. 1838; no. 7; pp. 1701 - 1709
• Main Authors: Escoffre, Jean-Michel, Bellard, Elisabeth, Faurie, Cécile, Sébaï, Sarra C., Golzio, Muriel, Teissié, Justin, Rols, Marie-Pierre
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2014; Elsevier
• Subjects: Adenosine Triphosphate - metabolism; Animals; Biotechnology; Cell Line; Cell Membrane - metabolism; Cell Membrane Permeability; Cell Survival - physiology; Cellular Biology; CHO Cells; Cricetulus; Electropermeabilization; Electroporation; Electroporation - methods; Life Sciences; Membrane disorder; Optical imaging; Phosphatidylcholines - metabolism; Phospholipid flip-flop; Phospholipids - metabolism; Electroporation; Optical imaging; Membrane disorder; Phospholipid flip-flop; Electropermeabilization
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/j.bbamem.2014.02.013

Membrane electropermeabilization relies on the transient permeabilization of the plasma membrane of cells submitted to electric pulses. This method is widely used in cell biology and medicine due to its efficiency to transfer molecules while limiting loss of cell viability. However, very little is known about the consequences of membrane electropermeabilization at the molecular and cellular levels. Progress in the knowledge of the involved mechanisms is a biophysical challenge. As a transient loss of membrane cohesion is associated with membrane permeabilization, our main objective was to detect and visualize at the single-cell level the incidence of phospholipid scrambling and changes in membrane order. We performed studies using fluorescence microscopy with C6-NBD-PC and FM1-43 to monitor phospholipid scrambling and membrane order of mammalian cells. Millisecond permeabilizing pulses induced membrane disorganization by increasing the translocation of phosphatidylcholines according to an ATP-independent process. The pulses induced the formation of long-lived permeant structures that were present during membrane resealing, but were not associated with phosphatidylcholine internalization. These pulses resulted in a rapid phospholipid flip/flop within less than 1s and were exclusively restricted to the regions of the permeabilized membrane. Under such electrical conditions, phosphatidylserine externalization was not detected. Moreover, this electrically-mediated membrane disorganization was not correlated with loss of cell viability. Our results could support the existence of direct interactions between the movement of membrane zwitterionic phospholipids and the electric field. Electro-induced phospholipid scrambling [Display omitted] •Electropermeabilization induces a rapid phosphatidylcholine translocation.•Phospholipid scrambling is restricted to the permeabilized regions.•Direct interactions between phospholipids and the electric field occurred.•Electrically-mediated membrane disorganization is not related to a loss of cell viability.