Lomax R B, Warhurst G, Sandle G I
Department of Medicine Gastrointestinal Molecular Physiology Group, (University of Manchester), Salford.
Gut. 1996 Feb;38(2):243-7. doi: 10.1136/gut.38.2.243.
The basolateral membrane of human colonic crypt cells contains Ca2+ and cAMP activated, Ba2+ blockable, low conductance (23 pS) K+ channels, which probably play an important part in intestinal Cl- secretion. This study has defined more clearly the basolateral K+ conductive properties of human colonic crypts using patch clamp recording techniques. High conductance (138 pS) K+ channels were seen in 25% of patches (one or two channels per patch), and significantly inhibited by the addition of 5 mM Ba2+, 1 mM quinidine or 20 mM tetraethylammonium chloride (TEA) to the cytosolic side of excised inside-out patches, whereas 1 mM diphenylamine-2-carboxylic acid (DPC) had no effect. In contrast, clusters of the 23 pS K+ channel (two to six channels per patch) were present in > 75% of patches, and channel activity was inhibited by quinidine and DPC, but not by TEA. Activity of the 138 pS K+ channel in inside-out patches was abolished almost completely by removal of bath Ca2+, but in contrast with its effect on the 23 pS K+ channel, addition of 0.1 mM carbachol had no effect on the 138 pS K+ channel in cell attached patches. It is concluded that human colonic crypt cells possess two discrete basolateral K+ channel populations, which can be distinguished by their responses to K+ channel blockers, and their different sensitivities to changes in intracellular Ca2+ concentration.
人结肠隐窝细胞的基底外侧膜含有Ca2+和cAMP激活、Ba2+可阻断、低电导(23 pS)的K+通道,这些通道可能在肠道Cl-分泌中起重要作用。本研究利用膜片钳记录技术更清楚地界定了人结肠隐窝的基底外侧K+传导特性。在25%的膜片中观察到高电导(138 pS)的K+通道(每个膜片一或两个通道),向切除的内向外膜片的胞质侧加入5 mM Ba2+、1 mM奎尼丁或20 mM四乙铵氯化物(TEA)可显著抑制该通道,而1 mM二苯胺-2-羧酸(DPC)则无作用。相比之下,>75%的膜片中存在23 pS K+通道簇(每个膜片两到六个通道),通道活性受奎尼丁和DPC抑制,但不受TEA抑制。去除浴液中的Ca2+几乎完全消除了内向外膜片中138 pS K+通道的活性,但与它对23 pS K+通道的作用相反,加入0.1 mM卡巴胆碱对细胞贴附膜片中的138 pS K+通道无作用。结论是人结肠隐窝细胞拥有两个离散的基底外侧K+通道群体,可通过它们对K+通道阻滞剂的反应以及对细胞内Ca2+浓度变化的不同敏感性来区分。