Kim C H, Rhee P L, Rhee J C, Kim Y I, So I, Kim K W, Park M K, Uhm D Y, Kang T M
Department of Physiology, Medicine and Surgery, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea.
Exp Physiol. 2000 Sep;85(5):497-504. doi: 10.1017/s0958067000020133.
The purpose of the present study was to characterize the Ca2+ channels in smooth muscle cells from human stomach and to examine the effects of osmotic swelling on the channel activity. Ca2+ channel current with either Ca2+ or Ba2+ as charge carrier was recorded from freshly isolated smooth muscle cells using the conventional whole-cell patch clamp technique. The degree of cell swelling as a result of hypotonic challenge was monitored using a video image analysis system. The changes in intracellular Ca2+ concentration ([Ca2+]i) were measured by microfluorimetry. The pharmacological and voltage activation profile suggests a typical dihydropyridine-sensitive L-type Ca2+ current. Cell swelling, induced by hypotonic challenge, enhanced the amplitude of currents through L-type Ca2+ channels without significant effects on steady-state voltage dependency. After treatment with the L-type Ca2+ channel agonist Bay K 8644 (0.1-2 microM), no further significant increase in calcium channel current or corresponding [Ca2+]i transients were provoked by the swelling. The above results demonstrated that the presence of L-type Ca2+ current in smooth muscle cells of the human stomach and the augmentation of the current are closely associated with the volume increase resulting from hypotonic swelling.
本研究的目的是对人胃平滑肌细胞中的钙离子通道进行表征,并研究渗透性肿胀对通道活性的影响。使用传统的全细胞膜片钳技术,从新鲜分离的平滑肌细胞中记录以钙离子或钡离子作为电荷载体的钙离子通道电流。使用视频图像分析系统监测低渗刺激导致的细胞肿胀程度。通过显微荧光测定法测量细胞内钙离子浓度([Ca2+]i)的变化。药理学和电压激活曲线表明存在典型的对二氢吡啶敏感的L型钙离子电流。低渗刺激诱导的细胞肿胀增强了通过L型钙离子通道的电流幅度,而对稳态电压依赖性没有显著影响。在用L型钙离子通道激动剂Bay K 8644(0.1 - 2 microM)处理后,肿胀不再引起钙离子通道电流或相应的[Ca2+]i瞬变的进一步显著增加。上述结果表明,人胃平滑肌细胞中存在L型钙离子电流,且该电流的增强与低渗肿胀导致的体积增加密切相关。