Mlinar B, Enyeart J J
Department of Pharmacology, Ohio State University, Columbus 43210-1239.
J Gen Physiol. 1993 Aug;102(2):239-55. doi: 10.1085/jgp.102.2.239.
In whole cell patch clamp recordings on enzymatically dissociated adrenal zona fasciculata (AZF) cells, a rapidly inactivating A-type K+ current was observed in each of more than 150 cells. Activation of IA was steeply voltage dependent and could be described by a Boltzmann function raised to an integer power of 4, with a midpoint of -28.3 mV. Using the "limiting logarithmic potential sensitivity," the single channel gating charge was estimated to be 7.2 e. Voltage-dependent inactivation could also be described by a Boltzmann function with a midpoint of -58.7 mV and a slope factor of 5.92 mV. Gating kinetics of IA included both voltage-dependent and -independent transitions in pathways between closed, open, and inactivated states. IA activated with voltage-dependent sigmoidal kinetics that could be fit with an n4h formalism. The activation time constant, tau a, reached a voltage-independent minimum at potentials positive to 0 mV. IA currents inactivated with two time constants that were voltage independent at potentials ranging from -30 to +45 mV. At +20 mV, tau i(fast) and tau i(slow) were 13.16 +/- 0.64 and 62.26 +/- 5.35 ms (n = 34), respectively. In some cells, IA inactivation kinetics slowed dramatically after many minutes of whole cell recording. Once activated by depolarization, IA channels returned to the closed state along pathways with two voltage-dependent time constants which were 0.208 s, tau rec-f and 10.02 s, tau rec-s at -80 mV. Approximately 90% of IA current recovered with slow kinetics at potentials between -60 and -100 mV. IA was blocked by 4-aminopyridine (IC50 = 629 microM) through a mechanism that was strongly promoted by channel activation. Divalent and trivalent cations including Ni2+ and La3+ also blocked IA with IC50's of 467 and 26.4 microM, respectively. With respect to biophysical properties and pharmacology, IA in AZF cells resembles to some extent transient K+ currents in neurons and muscle, where they function to regulate action potential frequency and duration. The function of this prominent current in steroid hormone secretion by endocrine cells that may not generate action potentials is not yet clear.
在对酶解肾上腺束状带(AZF)细胞进行的全细胞膜片钳记录中,在150多个细胞中的每一个细胞中都观察到一种快速失活的A型钾电流。IA的激活强烈依赖电压,可用幂次为4的玻尔兹曼函数描述,中点为-28.3 mV。使用“极限对数电位敏感性”,单通道门控电荷估计为7.2 e。电压依赖性失活也可用玻尔兹曼函数描述,中点为-58.7 mV,斜率因子为5.92 mV。IA的门控动力学包括在关闭、开放和失活状态之间的途径中的电压依赖性和非依赖性转变。IA以电压依赖性S形动力学激活,可用n4h形式拟合。激活时间常数tau a在正于0 mV的电位处达到与电压无关的最小值。IA电流以两个时间常数失活,在-30至+45 mV的电位范围内与电压无关。在+20 mV时,tau i(快)和tau i(慢)分别为13.16±0.64和62.26±5.35 ms(n = 34)。在一些细胞中,全细胞记录数分钟后,IA失活动力学显著减慢。一旦被去极化激活,IA通道沿具有两个电压依赖性时间常数的途径回到关闭状态,在-80 mV时,tau rec-f为0.208 s,tau rec-s为10.02 s。在-60至-100 mV的电位之间,约90%的IA电流以慢动力学恢复。4-氨基吡啶(IC50 = 629 microM)通过一种被通道激活强烈促进的机制阻断IA。包括Ni2+和La3+在内的二价和三价阳离子也阻断IA,IC50分别为467和26.4 microM。就生物物理特性和药理学而言,AZF细胞中的IA在一定程度上类似于神经元和肌肉中的瞬时钾电流,它们在其中发挥调节动作电位频率和持续时间的作用。这种在可能不产生动作电位的内分泌细胞类固醇激素分泌中突出电流的功能尚不清楚。