Gray M A, Greenwell J R, Argent B E
Department of Physiological Sciences, University Medical School, Newcastle upon Tyne, United Kingdom.
J Membr Biol. 1988 Oct;105(2):131-42. doi: 10.1007/BF02009166.
Using the patch clamp technique we have identified a small conductance ion channel that typically occurs in clusters on the apical plasma membrane of pancreatic duct cells. The cell-attached current/voltage (I/V) relationship was linear and gave a single channel conductance of about 4 pS. Since the reversal potential was close to the resting membrane potential of the cell, and unaffected by changing from Na+-rich to K+-rich pipette solutions, the channel selects for anions over cations in cell-attached patches. The open state probability was not voltage-dependent. Adding 25 mM-bicarbonate to the bath solution caused a slight outward rectification of the I/V relationship, but otherwise, the characteristics of the channel were unaffected. In excised, inside-out, patches the I/V relationship was linear and gave a single channel conductance of about 4 pS. A threefold chloride concentration gradient across the patch (sulphate replacement) shifted the single channel current reversal potential by -26 mV, indicating that the channel is chloride selective. Stimulation of duct cells with secretin (10 nM), dibutyryl cyclic AMP (1 mM) and forskolin (1 microM) increased channel open state probability and also increased the number of channels, and/or caused disaggregation of channel clusters, in the apical plasma membrane. Coupling of this channel to a chloride/bicarbonate exchanger would provide a mechanism for electrogenic bicarbonate secretion by pancreatic duct cells.
运用膜片钳技术,我们鉴定出一种小电导离子通道,该通道通常以簇状形式出现在胰腺导管细胞顶端质膜上。细胞贴附式电流/电压(I/V)关系呈线性,单通道电导约为4皮西门子。由于反转电位接近细胞的静息膜电位,且不受从富含Na⁺的移液管溶液更换为富含K⁺的移液管溶液的影响,该通道在细胞贴附式膜片中对阴离子的选择性高于阳离子。开放态概率不依赖电压。向浴液中添加25 mM碳酸氢盐会使I/V关系出现轻微外向整流,但除此之外,通道特性未受影响。在切除的内面向外膜片中,I/V关系呈线性,单通道电导约为4皮西门子。跨膜片三倍的氯离子浓度梯度(用硫酸盐替代)使单通道电流反转电位偏移了-26 mV,表明该通道具有氯离子选择性。用促胰液素(10 nM)、二丁酰环磷腺苷(1 mM)和福斯可林(1 microM)刺激导管细胞,可增加通道开放态概率,还可增加顶端质膜中通道的数量,和/或导致通道簇解聚。该通道与氯离子/碳酸氢盐交换器的偶联将为胰腺导管细胞的电生性碳酸氢盐分泌提供一种机制。