Thomas Dierk, Wu Kezhong, Wimmer Anna-Britt, Zitron Edgar, Hammerling Bettina C, Kathöfer Sven, Lueck Sonja, Bloehs Ramona, Kreye Volker A W, Kiehn Johann, Katus Hugo A, Schoels Wolfgang, Karle Christoph A
Department of Cardiology, Medical University Hospital, Im Neuenheimer Feld 410, 69120, Heidelberg, Germany.
J Mol Med (Berl). 2004 Dec;82(12):826-37. doi: 10.1007/s00109-004-0582-8. Epub 2004 Sep 8.
Patients with cardiac disease typically develop life-threatening ventricular arrhythmias during physical or emotional stress, suggesting a link between adrenergic stimulation and regulation of the cardiac action potential. Human ether-a-go-go related gene (hERG) potassium channels conduct the rapid component of the repolarizing delayed rectifier potassium current, I(Kr). Previous studies have revealed that hERG channel activation is modulated by activation of the beta-adrenergic system. In contrast, the influence of the alpha-adrenergic signal transduction cascade on hERG currents is less well understood. The present study examined the regulation of hERG currents by alpha(1A)-adrenoceptors. hERG channels and human alpha(1A)-adrenoceptors were heterologously coexpressed in Xenopus laevis oocytes, and currents were measured using the two-microelectrode voltage clamp technique. Stimulation of alpha(1A)-receptors by applying 20 microM phenylephrine caused hERG current reduction due to a 9.6-mV shift of the activation curve towards more positive potentials. Simultaneous application of the alpha(1)-adrenoceptor antagonist prazosin (20 microM) prevented the activation shift. Inhibition of PKC (3 microM Ro-32-0432) or PKA (2.5 microM KT 5720) abolished the alpha-adrenergic activation shift, suggesting that PKC and PKA are required within the regulatory mechanism. The effect was still present when the PKA- and PKC-dependent phosphorylation sites in hERG were deleted by mutagenesis. In summary, cardiac repolarizing hERG/I(Kr) potassium currents are modulated by alpha(1A)-adrenoceptors via PKC and PKA independently of direct channel phosphorylation. This novel regulatory pathway of alpha1-adrenergic hERG current regulation provides a link between stress and ventricular arrhythmias, in particular in patients with heart disease.
患有心脏疾病的患者通常会在身体或情绪应激期间出现危及生命的室性心律失常,这表明肾上腺素能刺激与心脏动作电位调节之间存在联系。人类醚 - 去极化相关基因(hERG)钾通道传导复极化延迟整流钾电流I(Kr)的快速成分。先前的研究表明,hERG通道激活受β - 肾上腺素能系统激活的调节。相比之下,α - 肾上腺素能信号转导级联对hERG电流的影响尚不太清楚。本研究检测了α(1A) - 肾上腺素能受体对hERG电流的调节作用。hERG通道和人类α(1A) - 肾上腺素能受体在非洲爪蟾卵母细胞中异源共表达,并使用双微电极电压钳技术测量电流。通过施加20μM去氧肾上腺素刺激α(1A) - 受体,导致hERG电流减少,这是由于激活曲线向更正电位方向偏移了9.6 mV。同时应用α(1) - 肾上腺素能受体拮抗剂哌唑嗪(20μM)可防止激活曲线偏移。抑制蛋白激酶C(3μM Ro - 32 - 0432)或蛋白激酶A(2.5μM KT 5720)可消除α - 肾上腺素能激活曲线偏移,表明在调节机制中需要蛋白激酶C和蛋白激酶A。当通过诱变删除hERG中依赖蛋白激酶A和蛋白激酶C的磷酸化位点时,该效应仍然存在。总之,心脏复极化hERG/I(Kr)钾电流受α(1A) - 肾上腺素能受体通过蛋白激酶C和蛋白激酶A调节,与直接通道磷酸化无关。这种α1 - 肾上腺素能对hERG电流调节的新途径为应激与室性心律失常之间提供了联系,尤其是在心脏病患者中。