1 The aim of the present study was to investigate the effects of 2,5-di-t-b
utyl-1,4-benzohydroquinone (BHQ), an inhibitor of the sarco-endoplasmic ret
iculum Ca2+-ATPase (SERCA), on the whole-cell voltage-dependent L-type Ca2 current of freshly isolated smooth muscle cells from the rat tail artery u
sing the patch-clamp technique.
2 BHQ, added to the perfusion solution, reduced I-Cat(L) in a concentration
- (IC50 = 66.7 muM) and voltage-dependent manner. This inhibition was only
partially reversible.
3 BHQ shifted the voltage dependence of the steady-state inactivation curve
to more negative potentials by 7 mV in the mid-potential of the curve, wit
hout affecting the activation curve as well as the time course of I-Ca(L) i
nactivation.
4 Preincubation of the cells either with 10 mum cyclopiazonic acid, a SERCA
inhibitor, or with 3 mm diethyldithiocarbamate, an inhibitor of intracellu
lar superoxide dismutase (SOD), did not modify BHQ inhibition of I-Ca(L). O
n the contrary, this effect was no longer evident when SOD (250 u ml(-1)) w
as added to the perfusion medium.
5 Either in the presence or in the absence of cells, BHQ gave rise to super
oxide anion formation, which was markedly inhibited by the addition of SOD.
6 These results indicate that, at micromolar concentrations, BHQ inhibits v
ascular by giving rise to the formation of superoxide anion which in turn i
mpairs the channel function.