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猪胰腺腺泡细胞中激素控制下钙激活钾通道的定量分析。

Quantification of Ca2+-activated K+ channels under hormonal control in pig pancreas acinar cells.

作者信息

Maruyama Y, Petersen O H, Flanagan P, Pearson G T

出版信息

Nature. 1983;305(5931):228-32. doi: 10.1038/305228a0.

Abstract

Ca2+- and voltage-activated K+ channels are found in many electrically excitable cells and have an important role in regulating electrical activity. Recently, the large K+ channel has been found in the baso-lateral plasma membranes of salivary gland acinar cells, where it may be important in the regulation of salt transport. Using patch-clamp methods to record single-channel currents from excised fragments of baso-lateral acinar cell membranes in combination with current recordings from isolated single acinar cells and two- and three-cell clusters, we have now for the first time characterized the K+ channels quantitatively. In pig pancreatic acini there are 25-60 K+ channels per cell with a maximal single channel conductance of about 200 pS. We have quantified the relationship between internal ionized Ca2+ concentration [( Ca2+]i) membrane potential and open-state probability (p) of the K+ channel. By comparing curves obtained from excised patches relating membrane potential to p, at different levels of [Ca2+]i, with similar curves obtained from intact cells, [Ca2+]i in resting acinar cells was found to be between 10(-8) and 10(-7) M. In microelectrode experiments acetylcholine (ACh), gastrin-cholecystokinin (CCK) as well as bombesin peptides evoked Ca2+-dependent opening of the K+ conductance pathway, resulting in membrane hyperpolarization. The large K+ channel, which is under strict dual control by internal Ca2+ and voltage, may provide a crucial link between hormone-evoked increase in internal Ca2+ concentration and the resulting NaCl-rich fluid secretion.

摘要

钙离子和电压激活的钾离子通道存在于许多电可兴奋细胞中,在调节电活动方面发挥着重要作用。最近,在唾液腺腺泡细胞的基底外侧质膜中发现了大钾离子通道,它在盐转运调节中可能具有重要作用。我们使用膜片钳方法记录基底外侧腺泡细胞膜切除片段的单通道电流,并结合分离的单个腺泡细胞以及双细胞和三细胞簇的电流记录,首次对钾离子通道进行了定量表征。在猪胰腺腺泡中,每个细胞有25 - 60个钾离子通道,最大单通道电导约为200 pS。我们已经量化了细胞内游离钙离子浓度[Ca2+]i、膜电位与钾离子通道开放概率(p)之间的关系。通过比较在不同[Ca2+]i水平下,从切除的膜片获得的膜电位与p的关系曲线,以及从完整细胞获得的类似曲线,发现静息腺泡细胞中的[Ca2+]i在10(-8)至10(-7) M之间。在微电极实验中,乙酰胆碱(ACh)、胃泌素 - 胆囊收缩素(CCK)以及蛙皮素肽均可诱发钙离子依赖的钾离子电导途径开放,导致膜超极化。这种受到细胞内钙离子和电压严格双重控制的大钾离子通道,可能在激素诱发的细胞内钙离子浓度增加与由此产生的富含氯化钠的液体分泌之间提供关键联系。

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