Kodirov S A, Zhuravlev V L, Pavlenko V K, Safonova T A, Brachmann J
Department of Cardiology, Medical University Hospital Heidelberg, 69120 Heidelberg, Germany.
J Membr Biol. 2004 Feb 1;197(3):145-54. doi: 10.1007/s00232-004-0649-z.
We used the patch-clamp technique to identify and characterize the electrophysiological, biophysical, and pharmacological properties of K(+) channels in enzymatically dissociated ventricular cells of the land pulmonate snail Helix. The family of outward K(+) currents started to activate at -30 mV and the activation was faster at more depolarized potentials (time constants: at 0 mV 17.4 +/- 1.2 ms vs. 2.5 +/- 0.1 ms at + 60 mV). The current waveforms were similar to those of the A-type family of voltage-dependent K(+) currents encoded by Kv4.2 in mammals. Inactivation of the current was relatively fast, i.e., 50.2 +/- 1.8% of current was inactivated within 250 ms at + 40 mV. The recovery of K(+) channels from inactivation was relatively slow with a mean time constant of 1.7 +/- 0.2 s. Closer examination of steady-state inactivation kinetics revealed that the voltage dependency of inactivation was U-shaped, exhibiting less inactivation at more depolarized membrane potentials. On the basis of this phenomenon, we suggest that a channel encoded by Kv2.1 similar to that in mammals does exist in land pulmonates of the Helix genus. Outward currents were sensitive to 4-aminopyridine and tetraethylammonium chloride. The last compound was most effective, with an IC(50) of 336 +/- 142 micro mol l(-1). Thus, using distinct pharmacological and biophysical tools we identified different types of voltage-gated K(+) channels.
我们运用膜片钳技术来鉴定和表征陆生肺螺类蜗牛Helix酶解心室细胞中钾离子通道的电生理、生物物理及药理学特性。外向钾电流家族在-30 mV时开始激活,且在更去极化的电位下激活更快(时间常数:在0 mV时为17.4±1.2 ms,在+60 mV时为2.5±0.1 ms)。电流波形与哺乳动物中由Kv4.2编码的电压依赖性A型钾电流家族的波形相似。电流的失活相对较快,即在+40 mV时,250 ms内有50.2±1.8%的电流失活。钾离子通道从失活状态恢复相对较慢,平均时间常数为1.7±0.2 s。对稳态失活动力学的进一步研究表明,失活的电压依赖性呈U形,在更去极化的膜电位下失活较少。基于这一现象,我们认为在Helix属的陆生肺螺类中确实存在一种与哺乳动物中类似的由Kv2.1编码的通道。外向电流对4-氨基吡啶和氯化四乙铵敏感。后一种化合物最为有效,IC50为336±142 μmol l-1。因此,我们使用不同的药理学和生物物理工具鉴定出了不同类型的电压门控钾离子通道。