Human embryonic kidney (HEK293) cells express endogenous voltage-gated sodium currents and Na v 1.7 sodium channels.
Human embryonic kidney (HEK293) cells are widely used for the heterologous expression of voltage- and ligand-gated ion channels. Patch clamp analysis of HEK293 cells in the whole-cell configuration identified voltage-gated, rapidly inactivating inward currents. Peak current amplitudes ranged from less than 100 pA to more than 800 pA, with the majority (84 of 130 cells) in the 100-400 pA range. Transient inward currents were separated into three components on the basis of sensitivity to cadmium and tetrodotoxin (TTX). Application of cadmium (300 microM) reduced current amplitude to 65% of control, consistent with the existence of current carried by a cadmium-sensitive nonspecific cation channel previously identified in HEK293 cells. Application of TTX (500 nM) reduced current amplitude by 47%, consistent with the existence of current carried by a TTX-sensitive voltage-gated sodium channel. Joint application of cadmium and TTX was additive, reducing current amplitude to 28% of control. The residual cadmium- and TTX-resistant currents represent a third pharmacologically distinct component of the rapidly inactivating inward current that was not characterized further. The pyrethroid insecticide tefluthrin (10 microM) prolonged the inactivation of transient currents and induced slowly decaying tail currents, effects that are characteristic of sodium channel modification by pyrethroids. The use of sodium channel isoform-specific primers in polymerase chain reaction amplifications on HEK293 cell first-strand cDNA detected the consistent expression of the human Na(v)1.7 sodium channel isoform in cells that expressed the TTX-sensitive component of current. These results provide evidence for an endogenous TTX-sensitive sodium current in HEK293 cells that is associated primarily with the expression of the Na(v)1.7 sodium channel isoform.
He
Bingjun
B
College of Life Sciences, Nankai University, Tianjin 300071, China.
Soderlund
David M
DM
D002104:Cadmium / Q000494:pharmacology; D002460:Cell Line; D003521:Cyclopropanes / Q000494:pharmacology; D006801:Humans; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects* / Q000502:physiology*; D049990:Membrane Transport Modulators / Q000494:pharmacology; D062556:NAV1.7 Voltage-Gated Sodium Channel; D018408:Patch-Clamp Techniques; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000378:metabolism; D026941:Sodium Channel Blockers / Q000494:pharmacology; D015222:Sodium Channels / Q000378:metabolism*; D013779:Tetrodotoxin / Q000494:pharmacology
D003521:Cyclopropanes; D006845:Hydrocarbons, Fluorinated; D049990:Membrane Transport Modulators; D062556:NAV1.7 Voltage-Gated Sodium Channel; D020033:Protein Isoforms; C486866:SCN9A protein, human; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D002104:Cadmium; C489827:2,3,5,6-tetrafluoro-4-methylbenzyl (Z)-(1RS)-cis-3-(2-chloro-3,3,3-trifluoroprop-1-enyl)-2,2-dimethylcyclopropanecarboxylate; D013779:Tetrodotoxin