Modulation of intracellular Ca2+ via L-type calcium channels in heart cells by the autoantibody directed against the second extracellular loop of the α1-adrenoceptors

Abstract

The effects of methoxamine, a selective α1-adrenergic receptor agonist, and the autoantibody directed against the second extracellular loop of α1-adrenoceptors were studied on intracellular free Ca2+ levels using confocal microscopy and ionic currents using the whole-cell patch clamp technique in single cells of 10-day-old embryonic chick and 20-week-old fetal human hearts. We observed that like methoxamine, the autoantibody directed against the second extracellular loop of α1-adrenoreceptors significantly increased the L-type calcium current (ICa(L)) but had no effect on the T-type calcium current (ICa(T)), the delayed outward potassium current, or the fast sodium current. This effect of the autoantibody was prevented by a prestimulation of the receptors with methoxamine and vice versa. Moreover, treating the cells with prazosin, a selective α1-adrenergic receptor antagonist blocked the methoxamine and the autoantibody-induced increase in ICa(L), respectively. In absence of prazosin, both methoxamine and the autoantibody showed a substantial enhancement in the frequency of cell contraction and that of the concomitant cytosolic and nuclear free Ca2+ variations. The subsequent addition of nifedipine, a specific L-type Ca2+ channel blocker, reversed not only the methoxamine or the autoantibody-induced effect but also completely abolished cell contraction. These results demonstrated that functional α1-adrenoceptors exist in both 10-day-old embryonic chick and 20-week-old human fetal hearts and that the autoantibody directed against the second extracellular loop of this type of receptors plays an important role in stimulating their activity via activation of L-type calcium channels. This loop seems to have a functional significance by being the target of α1-receptor agonists like methoxamine.Key words: α1-adrenoceptor, autoantibody, heart cells, calcium, ion channels, confocal microscopy.