Diffuse, non-polar electropermeabilization and reduced propidium uptake distinguish the effect of nanosecond electric pulses.

Semenov I, Zemlin C, Pakhomova ON, Xiao S, Pakhomov AG
Source: Other
Publication Date: (2015)
Issue: 1848(10A): 2118-2125
Research Area:
Cells used in publication:
Cardiomyocyte (R-CM), rat
Species: rat
Tissue Origin: heart
Ca2+ activation and membrane electroporation by 10-ns and 4-ms electric pulses (nsEP and msEP) were compared in rat embryonic cardiomyocytes. The lowest electric field which triggered Ca2+ transients was expectedly higher for nsEP (36kV/cm) than for msEP (0.09kV/cm) but the respective doses were similar (190 and 460mJ/g). At higher intensities, both stimuli triggered prolonged firing in quiescent cells. An increase of basal Ca2+ level by >10nM in cells with blocked voltage-gated Ca2+ channels and depleted Ca2+ depot occurred at 63kV/cm (nsEP) or 0.14kV/cm (msEP) and was regarded as electroporation threshold. These electric field values were at 150-230% of stimulation thresholds for both msEP and nsEP, notwithstanding a 400,000-fold difference in pulse duration. For comparable levels of electroporative Ca2+ uptake, msEP caused at least 10-fold greater uptake of propidium than nsEP, suggesting increased yield of larger pores. Electroporation by msEP started Ca2+ entry abruptly and locally at the electrode-facing poles of cell, followed by a slow diffusion to the center. In a stark contrast, nsEP evoked a "supra-electroporation" pattern of slower but spatially uniform Ca2+ entry. Thus nsEP and msEP had comparable dose efficiency, but differed profoundly in the size and localization of electropores.