Gardner P I
J Neurosci. 1986 Jul;6(7):2106-16. doi: 10.1523/JNEUROSCI.06-07-02106.1986.
Multiple distinct K+-selective channels may contribute to action potential repolarization and afterpotential generation in chick ciliary neurons. The channel types are difficult to distinguish by traditional voltage-clamp methods, primarily because of coactivation during depolarization. I have used the extracellular patch-clamp technique to resolve single-channel K+ currents in cultured chick ciliary ganglion (CG) neurons. Three unit currents selective for K+ ions were observed. The channels varied with respect to unit conductance, sensitivity to Ca2+ ions and voltage, and steady-state gating parameters. The first channel, GK1, was characterized by a unit conductance of 14 pico-Siemens (pS) under physiological recording conditions, gating that was relatively independent of membrane potential and intracellular Ca2+ ions, and single-component open-time distributions with time constants of approximately 9 msec. The second channel, GK2, was characterized by a unit conductance of 64 pS under physiological recording conditions and gating that was affected by membrane potential but was not dependent on the activity of intracellular Ca2+ ions. Open-time distributions indicated 2 open states, with open-time constants of 0.09 (61%) and 0.35 (39%) msec, at +40 mV membrane potential. The third channel, GKCa2+, was identified in isolated patch recordings in which the concentration of internal Ca2+ was 10(-7) M or greater, which was an absolute prerequisite for channel opening. GKCa2+ was characterized by a unit conductance of 193 pS in symmetrical 0.15 M KCl solutions, an open-state probability that was a function not only of [Ca2+]i, but also of membrane potential, and single-component open-time distribution with a time constant of 1.11 msec at -10 mV patch potential. These results suggest the presence of at least 3 distinct K+ channel populations in the membrane of cultured chick CG neurons.
多种不同的钾离子选择性通道可能参与雏鸡睫状神经元动作电位复极化和后电位的产生。这些通道类型很难用传统的电压钳方法区分,主要是因为在去极化过程中它们会共同激活。我采用细胞外膜片钳技术来解析培养的雏鸡睫状神经节(CG)神经元中的单通道钾电流。观察到三种对钾离子有选择性的单位电流。这些通道在单位电导、对钙离子和电压的敏感性以及稳态门控参数方面存在差异。第一种通道GK1,在生理记录条件下其单位电导为14皮西门子(pS),门控相对独立于膜电位和细胞内钙离子,单成分开放时间分布的时间常数约为9毫秒。第二种通道GK2,在生理记录条件下单位电导为64 pS,门控受膜电位影响,但不依赖于细胞内钙离子的活性。在膜电位为+40 mV时,开放时间分布显示有2个开放状态,开放时间常数分别为0.09(61%)和0.35(39%)毫秒。第三种通道GKCa2+,是在内部钙离子浓度为10^(-7) M或更高的分离膜片记录中鉴定出来的,这是通道开放的绝对前提条件。在对称的0.15 M KCl溶液中,GKCa2+的单位电导为193 pS,开放状态概率不仅是[Ca2+]i的函数,也是膜电位的函数,在膜片电位为-10 mV时单成分开放时间分布的时间常数为1.11毫秒。这些结果表明,在培养的雏鸡CG神经元膜中至少存在3种不同的钾通道群体。