Gray M A, Plant S, Argent B E
Department of Physiological Sciences, University Medical School, Newcastle upon Tyne, United Kingdom.
Am J Physiol. 1993 Mar;264(3 Pt 1):C591-602. doi: 10.1152/ajpcell.1993.264.3.C591.
Using the whole cell configuration of the patch-clamp technique, we have identified an adenosine 3',5'-cyclic monophosphate (cAMP)-regulated chloride conductance in pancreatic duct cells. Basal whole cell currents in single isolated cells were very low (approximately 5 pA/pF) but could be stimulated 17-fold by elevation of intracellular cAMP. The cAMP-activated currents exhibited 1) a high chloride selectivity, 2) a near linear current-voltage relationship, 3) time and voltage independence, 4) block by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) but not by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), and 5) an anion selectivity sequence based on permeability ratios of SCN > NO3 > Br > Cl > I > HCO3 > F > ClO4 > gluconate. Currents in single cells ran down within a few minutes; however, stable chloride currents could be recorded from duct cell clusters in which four or five cells were in electrical communication. We present evidence suggesting that these cAMP-regulated currents are carried by cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels. Physiologically, these CFTR channels act in parallel with chloride-bicarbonate exchangers to facilitate bicarbonate secretion across the apical plasma membrane of the duct cell.
运用膜片钳技术的全细胞模式,我们在胰腺导管细胞中鉴定出一种受3',5'-环磷酸腺苷(cAMP)调节的氯离子电导。单个分离细胞的基础全细胞电流非常低(约5 pA/pF),但细胞内cAMP升高可使其增大17倍。cAMP激活的电流表现出:1)对氯离子具有高度选择性;2)电流-电压关系近乎线性;3)与时间和电压无关;4)可被5-硝基-2-(3-苯丙基氨基)苯甲酸(NPPB)阻断,但不被4,4'-二异硫氰基芪-2,2'-二磺酸(DIDS)阻断;5)基于通透性比率的阴离子选择性顺序为SCN > NO3 > Br > Cl > I > HCO3 > F > ClO4 > 葡萄糖酸盐。单细胞中的电流在几分钟内就会衰减;然而,在四五个细胞存在电偶联的导管细胞簇中可记录到稳定的氯离子电流。我们提供的证据表明,这些受cAMP调节的电流是由囊性纤维化跨膜传导调节因子(CFTR)氯离子通道介导的。在生理情况下,这些CFTR通道与氯-碳酸氢根交换体协同作用,以促进碳酸氢根通过导管细胞顶端质膜的分泌。