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J Biol Chem, Vol. 273, Issue 33, 21084-21090, August 14, 1998
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
Human Saphenous Vein Endothelial Cells Express a
Tetrodotoxin-resistant, Voltage-gated Sodium Current
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
-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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