Velasco J M, Petersen O H
Biochim Biophys Acta. 1987 Jan 26;896(2):305-10. doi: 10.1016/0005-2736(87)90191-x.
Patch-clamp single-channel current recording experiments have been carried out on intact insulin-secreting RINm5F cells. Voltage-activation of high-conductance K+ channels were studied by selectively depolarizing the electrically isolated patch membrane under conditions with normal Ca2+ concentration in the bath solution but with or without Ca2+ in the patch pipette solution. When Ca2+ was present in the pipette, 40 mV to 120 mV depolarizing pulses (100 ms) from the normal resting potential (-70 mV) regularly evoked tetraethylammonium-sensitive large outward single-channel currents and the average open state probability during the pulses varied from about 0.015 (40 mV pulses) to 0.1 (120 mV pulses). In the absence of Ca2+ in the pipette solution the same protocol resulted in fewer and shorter K+ channel openings and the open-state probability varied from about 0.0015 (40 mV pulses) to about 0.03 (120 mV pulses). It is concluded that Ca2+ entering voltage-gated channels raises [Ca2+]i locally and thereby markedly enhances the open-state probability of tetraethylammonium-sensitive voltage-gated high-conductance K+ channels.
已在完整的胰岛素分泌细胞RINm5F上进行了膜片钳单通道电流记录实验。通过在浴槽溶液中Ca2+浓度正常但膜片吸管溶液中有或无Ca2+的条件下选择性地使电隔离的膜片去极化,研究了高电导K+通道的电压激活。当吸管中有Ca2+时,从正常静息电位(-70 mV)开始的40 mV至120 mV去极化脉冲(100 ms)通常会诱发四乙铵敏感的大外向单通道电流,脉冲期间的平均开放状态概率从约0.015(40 mV脉冲)变化到0.1(120 mV脉冲)。在吸管溶液中无Ca2+的情况下,相同的方案导致K+通道开放次数减少且持续时间缩短,开放状态概率从约0.0015(40 mV脉冲)变化到约0.03(120 mV脉冲)。结论是,进入电压门控通道的Ca2+会局部升高[Ca2+]i,从而显著提高四乙铵敏感的电压门控高电导K+通道的开放状态概率。