Silencing of ATP1A1 attenuates cell membrane disruption by nanosecond electric pulses

• DOI: 10.1016/j.bbrc.2023.08.011
• Publication Date: 2023-08-05T01:42:13Z
• Authors: Giedre Silkuniene; Uma M. Mangalanathan; Andrei G. Pakhomov; Olga N. Pakhomova
• Keywords: Cell Membrane Permeability; Electroporation; Electricity; Cell Membrane; Humans; RNA, Small Interfering; Sodium-Potassium-Exchanging ATPase

This study explored the role of the Na/K-ATPase (NKA) in membrane permeabilization induced by nanosecond electric pulses. Using CRISPR/Cas9 and shRNA, we silenced the ATP1A1 gene, which encodes α1 NKA subunit in U937 human monocytes. Silencing reduced the rate and the cumulative uptake of YoPro-1 dye after electroporation by 300-ns, 7-10 kV/cm pulses, while ouabain, a specific NKA inhibitor, enhanced YoPro-1 entry. We conclude that the α1 subunit supports the electropermeabilized membrane state, by forming or stabilizing electropores or by hindering repair mechanisms, and this role is independent of NKA's ion pump function.