GM1 asymmetry in the membrane stabilizes pores

Cell membranes are highly asymmetric and their stability against poration is crucial for survival. We investigated the influence of membrane asymmetry on electroporation of giant unilamellar vesicles with membranes doped with GM1, a ganglioside asymmetrically enriched in the outer leaflet of neuronal cell membranes. Compared to symmetric membranes, the lifetimes of micronsized pores are about an order of magnitude longer suggesting that pores are stabilized by GM1. Internal membrane nanotubes caused by the GM1 asymmetry, obstruct and additionally slow down pore closure, effectively reducing pore edge tension and leading to leaky membranes. Our results point to the drastic effects this ganglioside can have on pore resealing in biotechnology applications based on poration as well as on membrane repair processes. SIGNIFICANCEMembrane pore closure is crucial for cell survival and is important for biotechnological and medicine applications based on transfer of material, e.g. drugs, genes, through pores. Electroporation is widely used as means to perforate the membrane but factors governing membrane resealing are still a matter of debate, in particular the large variations of pore lifetimes. Here, we probed the effect of bilayer asymmetry on pore dynamics employing cell-sized giant unilamellar vesicles doped with the ganglioside GM1 (asymmetrically enriched in neurons). We find that the presence of GM1 and its asymmetric distribution in the bilayer dramatically slows down pore resealing, not only by mere molecular stabilization of the pore rim, but also by generating membrane nanotubes.