Mitchell C H, Jacob T J
Eye Research Lab, School of Molecular and Medical Bioscience, University of Wales, Cardif, UK.
J Membr Biol. 1996 Mar;150(1):105-11. doi: 10.1007/s002329900034.
An ion channel activated by hyperpolarization was identified in excised patches of bovine pigmented ciliary epithelial cells using the single channel patch clamp technique. In symmetrical NaGluconate, the channel had a maximum conductance of 285 pS. The channel was characterized by frequent flickery transitions between the fully open and closed levels. The channel did not discriminate very clearly between anions and cations; when the cytoplasmic face of excised patches was bathed in a dilute NaCl solution, the channel had a chloride-to-sodium permeability ratio (PCl/PNa) of 1.3. However, the channel showed a small anion selectivity (PCl/PNa = 3.7) when bathed in a concentrated NaCl solution. Gd3+ blocked the channel reversibly. Channel kinetics were characterised by slow (approximately min) voltage-dependent activation and inactivation rate constants. The channel was most active in the range-60 to -140 mV and showed a peak at -120 mV. A similar time- and voltage-dependent activation was also observed in cell-attached recordings. In conclusion, hyperpolarization of pigmented ciliary epithelial cell membrane patches activated a large conductance, nonselective ion channel. This combination of nonselectivity and hyperpolarizing activation is consistent with the involvement of this channel in ion loading from the blood into pigmented ciliary epithelial cells-the first phase in the secretion of aqueous humor.
利用单通道膜片钳技术,在牛色素性睫状上皮细胞的膜片上鉴定出一种由超极化激活的离子通道。在对称的葡萄糖酸钠溶液中,该通道的最大电导为285 pS。该通道的特征是在完全开放和关闭状态之间频繁发生闪烁式转变。该通道对阴离子和阳离子的区分不是很明显;当膜片的胞质面浸泡在稀氯化钠溶液中时,该通道的氯钠通透率(PCl/PNa)为1.3。然而,当浸泡在浓氯化钠溶液中时,该通道表现出较小的阴离子选择性(PCl/PNa = 3.7)。钆离子(Gd3+)可可逆地阻断该通道。通道动力学的特征是缓慢(约数分钟)的电压依赖性激活和失活速率常数。该通道在-60至-140 mV范围内活性最高,在-120 mV时出现峰值。在细胞贴附记录中也观察到了类似的时间和电压依赖性激活。总之,色素性睫状上皮细胞膜片的超极化激活了一种大电导、非选择性的离子通道。这种非选择性和超极化激活的组合与该通道参与从血液向色素性睫状上皮细胞的离子装载过程一致,这是房水分泌的第一阶段。