Watanabe S I, Tanizaki M, Kaneko A
Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Exp Eye Res. 1997 Jun;64(6):1027-35. doi: 10.1006/exer.1997.0293.
Ion channels contribute to the regulation of cellular function through control of the membrane potential and intracellular concentration of various ions. We examined stretch-activated channels in the corneal epithelial cell. Patch clamping was applied to enzymatically dissociated corneal epithelial cells to characterize their stretch-activated ion channels. The plasma membrane was stretched by applying suction to the patch pipette in cell-attached or inside-out patch configuration. The ion selectivity, voltage-dependence, and stretch-dependence were examined. Two kinds of stretch-activated channel events were observed; the previously-reported large conductance (L) channel and a novel small conductance (S) channel. The probability of recording L vs. S channels in the cell-attached configuration was about 2:1. The L channel was potassium selective with single channel conductance (gamma) of about 160 pS under the symmetrical (150 mm K+) solution. The S channel was permeable to Na+ and K+ with gamma of about 20 pS under the same conditions. Both L and S channels showed little activity in the absence of suction applied to the recording pipette. Channel activity was evoked by suction (negative pressure) stronger than -20 mmHg in both channels. The open probability (Po) and the mean current increased in proportion to further applied stretch and did not saturate for applied suction as strong as -80 mmHg, the pressure at which the gigaseal started to break. Thus, two types of stretch-activated channels coexist in corneal epithelial cells; a potassium-selective L channel and non-selective S channel. The contribution of these channels to the membrane potential is discussed.
离子通道通过控制膜电位和各种离子的细胞内浓度来调节细胞功能。我们研究了角膜上皮细胞中的牵张激活通道。采用膜片钳技术对酶解分离的角膜上皮细胞进行检测,以表征其牵张激活离子通道。在细胞贴附式或内面向外式膜片配置中,通过对膜片吸管施加吸力来拉伸质膜。检测了离子选择性、电压依赖性和牵张依赖性。观察到两种牵张激活通道事件;先前报道的大电导(L)通道和一种新型的小电导(S)通道。在细胞贴附式配置中记录L通道与S通道的概率约为2:1。在对称(150 mM K+)溶液下,L通道对钾离子具有选择性,单通道电导(γ)约为160 pS。在相同条件下,S通道对Na+和K+通透,γ约为20 pS。在未对记录吸管施加吸力时,L通道和S通道的活性都很低。两种通道在施加强度大于 -20 mmHg的吸力(负压)时都会诱发通道活性。开放概率(Po)和平均电流与进一步施加的牵张成比例增加,并且对于高达 -80 mmHg的施加吸力(千兆封接开始破裂时的压力)不会饱和。因此,两种类型的牵张激活通道共存于角膜上皮细胞中;一种钾离子选择性L通道和非选择性S通道。讨论了这些通道对膜电位的贡献。