Ohya Susumu, Asakura Keiichi, Muraki Katsuhiko, Watanabe Minoru, Imaizumi Yuji
Department of Molecular and Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabedori, Mizuhoku, Nagoya 467-8603, Japan.
Am J Physiol Gastrointest Liver Physiol. 2002 Feb;282(2):G277-87. doi: 10.1152/ajpgi.00200.2001.
Contribution of K(+) channels derived from the expression of ERG, KCNQ, and KCNE subtypes, which are responsible for rapidly and slowly activating delayed rectifier K(+) currents (I(Kr) and I(Ks), respectively) in cardiac myocytes, to membrane currents was examined in stomach circular smooth muscle cells (SMCs). The region-qualified multicell RT-PCR showed that ERG1/KCNE2 transcripts were expressed in rat stomach fundus and antrum SMCs and that KCNQ1/KCNE1 transcripts were expressed in antrum but not fundus. Western blotting and immunocytochemical analyses indicate that ERG1 proteins were substantially expressed in both regions, whereas KCNE1 proteins were faintly expressed in antrum and not in fundus. Both I(Kr)- and I(Ks)-like currents susceptible to E-4031 and indapamide, respectively, were identified in circular SMCs of antrum but only I(Kr)-like current was identified in fundus. It is strongly suggested that I(Kr)- and I(Ks)-like currents functionally identified in rat stomach SMCs are attributable to the expression of ERG1/KCNE2 and KCNQ1/KCNE1, respectively. The membrane depolarization by 1 microM E-4031 indicates the contribution of K(+) channels encoded by ERG1/KCNE2 to the resting membrane potential in stomach SMCs.
在胃环形平滑肌细胞(SMC)中,研究了由ERG、KCNQ和KCNE亚型表达产生的钾通道对膜电流的贡献,这些亚型分别负责心肌细胞中快速和缓慢激活的延迟整流钾电流(分别为I(Kr)和I(Ks))。区域特异性多细胞RT-PCR显示,ERG1/KCNE2转录本在大鼠胃底和胃窦SMC中表达,而KCNQ1/KCNE1转录本在胃窦而非胃底表达。蛋白质印迹和免疫细胞化学分析表明,ERG1蛋白在两个区域均大量表达,而KCNE1蛋白在胃窦中微弱表达,在胃底中不表达。在胃窦的环形SMC中分别鉴定出对E-4031和吲达帕胺敏感的I(Kr)样电流和I(Ks)样电流,但在胃底仅鉴定出I(Kr)样电流。强烈提示,在大鼠胃SMC中功能鉴定出的I(Kr)样电流和I(Ks)样电流分别归因于ERG1/KCNE2和KCNQ1/KCNE1的表达。1 microM E-4031引起的膜去极化表明ERG1/KCNE2编码的钾通道对胃SMC静息膜电位有贡献。