Key result
Expression of alpha(1E) with beta(2b) and alpha(2)/delta in rabbit ventricular myocytes significantly increased total Ca(2+) current, generating a high-voltage activated, PN-resistant current.
Expression of alpha1E in cardiac myocytes generates a high-voltage activated calcium current rather than a T-like current, and modulates intrinsic L-type currents, likely through competition for auxiliary beta subunits.
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Gene expression alters native cardiac Ca currents; leaves open specificity of transfected alpha(1E) currents in myocytes.
Using the whole-cell patch-clamp technique, we have studied the properties of alpha(1E) Ca(2+) channel transfected in cardiac myocytes. We have also investigated the effect of foreign gene expression on the intrinsic L-type current (I(Ca,L)). Expression of green fluorescent protein significantly decreased the I(Ca,L). By contrast, expression of alpha(1E) with beta(2b) and alpha(2)/delta significantly increased the total Ca(2+) current, and in these cells a Ca(2+) antagonist, PN-200-110 (PN), only partially blocked the current. The remaining PN-resistant current was abolished by the application of a low concentration of Ni(2+) and was little affected by changing the charge carrier from Ca(2+) to Ba(2+) or by beta-adrenergic stimulation. On the basis of its voltage range for activation, this channel was classified as a high-voltage activated channel. Thus the expression of alpha(1E) did not generate T-like current in cardiac myocytes. On the other hand, expression of alpha(1E) decreased I(Ca,L) and slowed the I(Ca,L) inactivation. This inactivation slowing was attenuated by the beta(2b) coexpression, suggesting that the alpha(1E) may slow the inactivation of I(Ca,L) by scrambling with alpha(1C) for intrinsic auxiliary beta.
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Tateyama et al. (2001) studied this question. Expression of alpha(1E) Ca(2+) channel with beta(2b) and alpha(2)/delta vs. Expression of green fluorescent protein was evaluated on Total Ca(2+) current and I(Ca,L) properties. Expression of alpha(1E) with beta(2b) and alpha(2)/delta in rabbit ventricular myocytes significantly increased total Ca(2+) current, generating a high-voltage activated, PN-resistant current.
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