Xu C, Tang G, Lu Y, Wang R
Department of Physiology, University of Saskatchewan, Saskatoon, Canada.
Life Sci. 2000 Apr 14;66(21):2023-33. doi: 10.1016/s0024-3205(00)00529-4.
The molecular basis of voltage-dependent K+ (Kv) current in smooth muscle cells (SMCs) from rat tail artery was investigated by screening transcriptional expression of 15 Kv channel alpha-subunits and 3 Kv beta-subunits using RT-PCR technique. Among Kv genes that encode delayed rectifier Kv currents, mRNAs of Kv1.2, Kv1.3, Kv1.5, Kv2.1, Kv2.2, and Kv3.2 were expressed, but those of Kv1.1, Kv1.6, and Kv3.1 were not detected. The transient outward Kv current-encoding genes Kv1.4, Kv3.3, Kv3.4, Kv4.1- Kv4.3 as well as Kvbeta1, Kvbeta2, and Kvbeta3 were also expressed at mRNA level. Western blot study demonstrated the presence of Kv1.2, Kv1.3, Kv1.5, and Kv2.1, but not Kv3.2 proteins, in tail artery tissue. Immunocytochemistry study confirmed the presence of Kv1.2, Kv1.3, Kv1.5, and Kv2.1 channel proteins in primary cultured single SMCs. Our results represent the first systematic characterization of Kv gene expression in rat tail artery SMCs.
通过使用逆转录聚合酶链反应(RT-PCR)技术筛选15种电压依赖性钾离子(Kv)通道α亚基和3种Kvβ亚基的转录表达,研究了大鼠尾动脉平滑肌细胞(SMC)中电压依赖性K⁺(Kv)电流的分子基础。在编码延迟整流Kv电流的Kv基因中,Kv1.2、Kv1.3、Kv1.5、Kv2.1、Kv2.2和Kv3.2的信使核糖核酸(mRNA)有表达,但未检测到Kv1.1、Kv1.6和Kv3.1的mRNA。编码瞬时外向Kv电流的基因Kv1.4、Kv3.3、Kv3.4、Kv4.1 - Kv4.3以及Kvβ1、Kvβ2和Kvβ3在mRNA水平也有表达。蛋白质印迹研究表明,尾动脉组织中存在Kv1.2、Kv1.3、Kv1.5和Kv2.1蛋白,但不存在Kv3.2蛋白。免疫细胞化学研究证实,原代培养的单个SMC中存在Kv1.2、Kv1.3、Kv1.5和Kv2.1通道蛋白。我们的结果首次系统地表征了大鼠尾动脉SMC中Kv基因的表达。