Harhun Maksym I, Jurkiewicz Aron, Jurkiewicz Neide H, Kryshtal Dmytro O, Shuba Michael F, Vladimirova Irina A
Nerve-Muscle Physiology Department, Bogomoletz Institute of Physiology, Bogomoletz Street 4, Kiev-24, Ukraine.
Pflugers Arch. 2003 Jun;446(3):380-6. doi: 10.1007/s00424-002-0986-7. Epub 2003 Apr 4.
Voltage-gated components of the outward current in single smooth muscle cells isolated from the epididymal part of the rat vas deferens were studied using amphotericin B perforated patch-clamp techniques. The complex kinetics of the net outward current elicited by positive voltage steps from -80 mV to +40 mV suggested the presence of several components. Bath application of 200 nM charybdotoxin, a potent blocker of large-conductance, Ca(2+)-dependent K(+) channels (BK(Ca)), reduced the current amplitude significantly. When BK(Ca) channels were suppressed, fast-inactivating (I(K,f)) and delayed rectifying (I(K,dr)) components of the outward current were identified. I(K,f) was characterized by fast kinetics of current decay, negative steady-state activation and inactivation dependencies and sensitivity to 4-aminopyridine with an apparent K(d) of 0.32 mM, properties similar to those of the A-type K(+) current. In contrast, I(K,dr) activated and inactivated at more positive potentials. The time constant of activation of I(K,dr) was voltage dependent with an e-fold decrease per 21 mV depolarization. I(K,dr) was inhibited by clofilium, a blocker of voltage-gated K(+) channels, with an IC(50) of 12 micro M and was not blocked by 5 mM 4-aminopyridine. The possible significance of the voltage-gated currents is discussed.
运用两性霉素B穿孔膜片钳技术,研究了从大鼠输精管附睾部分分离出的单个平滑肌细胞外向电流的电压门控成分。从-80mV到+40mV的正电压阶跃所引发的净外向电流的复杂动力学表明存在几种成分。向浴槽中加入200 nM的大电导钙依赖性钾通道(BK(Ca))强效阻滞剂蝎毒素,可显著降低电流幅度。当BK(Ca)通道被抑制时,可识别出外向电流的快速失活成分(I(K,f))和延迟整流成分(I(K,dr))。I(K,f)的特征为电流衰减动力学快、稳态激活和失活依赖性为负以及对4-氨基吡啶敏感,其表观解离常数(K(d))为0.32 mM,这些特性与A 型钾电流相似。相比之下,I(K,dr)在更正的电位下激活和失活。I(K,dr)的激活时间常数与电压有关,每去极化21 mV下降一个数量级。I(K,dr)被电压门控钾通道阻滞剂氯非铵抑制,半数抑制浓度(IC(50))为12 μM,且不被5 mM 4-氨基吡啶阻断。文中讨论了电压门控电流的可能意义。