Miyashita T, Tatsumi H, Furuta H, Mori N, Sokabe M
Department of Otolaryngology, Kagawa Medical University, Kagawa 761-0793, Japan.
J Membr Biol. 2001 Jul 15;182(2):113-22. doi: 10.1007/s00232-001-0035-z.
We identified a Ca2+-sensitive cation channel in acutely dissociated epithelial cells from the endolymphatic sac (ES) of guinea pigs using the patch-clamp technique. Single-channel recordings showed that the cation channel had a conductance of 24.0 +/- 1.3 pS (n = 8) in our standard solution. The relative ionic permeability of the channel was in the order K+ = Na+ > Ca2+ >> Cl-. This channel was weakly voltage-dependent but was strongly activated by Ca2+ on the cytosolic side at a concentration of around 1 mm in inside-out excised patches. With cell-attached patches, however, the channel was activated by much lower Ca2+ concentrations. Treatment of the cells, under cell-attached configuration, with ionomycin (10 microm), carbonyl cyanide 3-chlorophenylhydrazone (CCCP, 20 microm), or ATP (1 mm), which increased intracellular Ca2+ concentration ([Ca2+]i), activated the channel at an estimated [Ca2+]i from 0.6 microm to 10 microm. It is suggested that some activators of the channel were deteriorated or washed out during the formation of excised patches. Based on this Ca2+ sensitivity, we speculated that the channel contributes to the regulation of ionic balance and volume of the ES by absorbing Na+ under certain pathological conditions that will increase [Ca2+]i. This is the first report of single-channel recordings in endolymphatic sac epithelial cells.
我们使用膜片钳技术在豚鼠内淋巴囊(ES)急性分离的上皮细胞中鉴定出一种Ca2+敏感阳离子通道。单通道记录显示,在我们的标准溶液中,该阳离子通道的电导为24.0±1.3 pS(n = 8)。该通道的相对离子通透性顺序为K+ = Na+ > Ca2+ >> Cl-。此通道电压依赖性较弱,但在膜外翻式膜片内,胞质侧Ca2+浓度约为1 mM时可被强烈激活。然而,在细胞贴附式膜片中,该通道可被低得多的Ca2+浓度激活。在细胞贴附式配置下,用离子霉素(10 μM)、羰基氰3-氯苯腙(CCCP,20 μM)或ATP(1 mM)处理细胞,这些物质会增加细胞内Ca2+浓度([Ca2+]i),在估计的[Ca2+]i为0.6 μM至10 μM时激活该通道。提示在膜片形成过程中,该通道的一些激活剂会降解或被洗脱。基于这种Ca2+敏感性,我们推测在某些会增加[Ca2+]i的病理条件下,该通道通过吸收Na+有助于调节内淋巴囊的离子平衡和体积。这是关于内淋巴囊上皮细胞单通道记录的首次报道。