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This study explored if the inhibition of VG Na<jats:sup>+<\/jats:sup> current (<jats:italic>I<\/jats:italic><jats:sub>Na<\/jats:sub>) resulted from (i) reduction of the transmembrane Na<jats:sup>+<\/jats:sup> gradient due to its influx via nsPEF\u2010opened pores, and\/or (ii) downregulation of the VG channels by a Ca<jats:sup>2+<\/jats:sup>\u2010dependent mechanism. We found that a single 300\u2009ns electric pulse at 1.6\u20135.3\u2009kV\/cm triggered sustained Na<jats:sup>+<\/jats:sup> influx in exposed NG108 cells and in primary chromaffin cells, as detected by increased fluorescence of a Sodium Green Dye. In the whole\u2010cell patch clamp configuration, this influx was efficiently buffered by the pipette solution so that the increase in the intracellular concentration of Na<jats:sup>+<\/jats:sup> ([Na]<jats:sub><jats:italic>i<\/jats:italic><\/jats:sub>) did not exceed 2\u20133\u2009mM. [Na]<jats:sub><jats:italic>i<\/jats:italic><\/jats:sub> increased uniformly over the cell volume and showed no additional peaks immediately below the plasma membrane. Concurrently, nsPEF reduced VG <jats:italic>I<\/jats:italic><jats:sub>Na<\/jats:sub> by 30\u201360% (at 4 and 5.3\u2009kV\/cm). In control experiments, even a greater increase of the pipette [Na<jats:sup>+<\/jats:sup>] (by 5\u2009mM) did not attenuate VG <jats:italic>I<\/jats:italic><jats:sub>Na<\/jats:sub>, thereby indicating that the nsPEF\u2010induced Na<jats:sup>+<\/jats:sup> influx was not the cause of VG <jats:italic>I<\/jats:italic><jats:sub>Na<\/jats:sub> inhibition. Similarly, adding 20\u2009mM of a fast Ca<jats:sup>2+<\/jats:sup> chelator 1,2\u2010bis(<jats:italic>o<\/jats:italic>\u2010aminophenoxy)ethane\u2010<jats:italic>N<\/jats:italic>,<jats:italic>N<\/jats:italic>,<jats:italic>N<\/jats:italic>\u2032,<jats:italic>N<\/jats:italic>\u2032\u2010tetraacetic acid (BAPTA) into the pipette solution did not prevent or attenuate the inhibition of the VG <jats:italic>I<\/jats:italic><jats:sub>Na<\/jats:sub> by nsPEF. These findings point to possible Ca<jats:sup>2+<\/jats:sup>\u2010independent downregulation of the VG Na<jats:sup>+<\/jats:sup> channels (e.g., caused by alteration of the lipid bilayer) or the direct effect of nsPEF on the channel. Bioelectromagnetics  33:443\u2013451, 2012. \u00a9 2012 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/bem.21703","type":"journal-article","created":{"date-parts":[[2012,1,10]],"date-time":"2012-01-10T13:17:41Z","timestamp":1326201461000},"page":"443-451","source":"Crossref","is-referenced-by-count":46,"title":["Inhibition of voltage\u2010gated Na<sup>+<\/sup> current by nanosecond pulsed electric field (nsPEF) is not mediated by Na<sup>+<\/sup> influx or Ca<sup>2+<\/sup> signaling"],"prefix":"10.1002","volume":"33","author":[{"given":"Vasyl","family":"Nesin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrei G.","family":"Pakhomov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2012,1,10]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.187559"},{"key":"e_1_2_6_3_1","unstructured":"BowmanA XiaoS SchoenbachKS PakhomovAG.2008.Inhibition of voltage\u2010gated calcium channels of cell plasma membrane by nanosecond electric pulses. 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