Pore resealing inactivation in electroporated erythrocyte membrane irradiated with electrons

The changes in the electroporation process of human erythrocytes membrane due to the direct action of high energy electron radiation were investigated. To avoid the indirect effects caused by radiolytic products of water, the irradiation was performed at liquid nitrogen temperature. The irradiated cells have been exposed to square-wave electric pulses at 4°C in isotonic suspensions to induce membrane electropores. The pores resealing were quantified by monitoring the cell hemolysis. A significant decrease of the resealing process was found for irradiation doses higher than 100 Gy. The mass of molecular structures affected by the direct action of radiation was estimated using the target analysis method. We found a molecular weight M_m≈930 kDa roughly corresponding to spectrin tetramer of the cytoskeleton. This suggests that spectrin network plays an important role in the pores resealing of the electropermeabilized erythrocyte membrane.