Rorsman P, Trube G
Pflugers Arch. 1985 Dec;405(4):305-9. doi: 10.1007/BF00595682.
The resting conductance of cultured beta-cells from murine pancreases was investigated using the whole-cell, cell-attached and isolated patch modes of the patch-clamp technique. Whole-cell experiments revealed a high input resistance of the cells (greater than 20 G omega per cell or greater than 100 k omega X cm2), if the medium dialysing the cell interior contained 3 mM ATP. The absence of ATP evoked a large additional K+ conductance. In cell-attached patches single K+-channels were observed in the absence of glucose. Addition of glucose (20 mM) to the bath suppressed the channel activity and initiated action potentials. Similar single-channel currents were recorded from isolated patches. In this case the channels were reversibly blocked by adding ATP (3 mM) to the solution at the intracellular side of the membrane. The conductances (51 pS and 56 pS for [K+]0 = 145 mM, T = 21 degrees C) and kinetics (at -70 mV: tau open = 2.2 ms and 1.8 ms, tau closed = 0.38 ms and 0.33 ms) of the glucose- and ATP-dependent channels were found to be very similar. It is concluded that both channels are identical. The result suggests that glucose could depolarize the beta-cell by increasing the cytoplasmic concentration of ATP.
采用膜片钳技术的全细胞、细胞贴附及单通道模式,对来自小鼠胰腺的培养β细胞的静息电导进行了研究。全细胞实验显示,如果透析细胞内液的培养基含有3 mM ATP,细胞具有高输入电阻(每个细胞大于20 GΩ或大于100 kΩ×cm2)。ATP缺失会引发大量额外的K+电导。在细胞贴附膜片中,在无葡萄糖的情况下观察到单个K+通道。向浴液中加入葡萄糖(20 mM)可抑制通道活性并引发动作电位。从分离的膜片中记录到类似的单通道电流。在这种情况下,通过向膜内侧的溶液中加入ATP(3 mM),通道被可逆性阻断。发现葡萄糖依赖性和ATP依赖性通道的电导(在[K+]0 = 145 mM、T = 21℃时分别为51 pS和56 pS)和动力学(在-70 mV时:开放时间常数tau open = 2.2 ms和1.8 ms,关闭时间常数tau closed = 0.38 ms和0.33 ms)非常相似。得出结论,两种通道是相同的。结果表明,葡萄糖可通过增加细胞质中ATP的浓度使β细胞去极化。