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J Biol Chem, Vol. 273, Issue 33, 21084-21090, August 14, 1998

Human Saphenous Vein Endothelial Cells Express a Tetrodotoxin-resistant, Voltage-gated Sodium Current

Martin Gosling, Suzanne L. Harley, Robert J. Turner, Nessa Carey, and Janet T. Powell

From the Department of Vascular Surgery, Imperial College School of Medicine at Charing Cross, Charing Cross Hospital, Fulham Palace Road, London W6 8RF, United Kingdom

Whole-cell patch-clamp electrophysiological investigation of endothelial cells cultured from human saphenous vein (HSVECs) has identified a voltage-gated Na+ current with a mean peak magnitude of -595 ± 49 pA (n = 75). This current was inhibited by tetrodotoxin (TTX) in a concentration-dependent manner, with an IC50 value of 4.7 µM, suggesting that it was of the TTX-resistant subtype. An antibody directed against the highly conserved intracellular linker region between domains III and IV of known Na+ channel alpha -subunits was able to retard current inactivation when applied intracellularly. This antibody identified a 245-kDa protein from membrane lysates on Western blotting and positively immunolabeled both cultured HSVECs and intact venous endothelium. HSVECs were also shown by reverse transcription-polymerase chain reaction to contain transcripts of the hH1 sodium channel gene. The expression of Na+ channels by HSVECs was shown using electrophysiology and cell-based enzyme-linked immunosorbent assay to be dependent on the concentration and source of human serum. Together, these results suggest that TTX-resistant Na+ channels of the hH1 isoform are expressed in human saphenous vein endothelium and that the presence of these channels is controlled by a serum factor.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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