Whole-cell Na+-activated K+ (KNa) channel currents and single KNa channels were studied with the patch-clamp method in small (20-25 micrometer) dorsal root ganglion (DRG) neurones in slices of rat dorsal root ganglia. 2. The whole-cell KNa channel current was identified as an additional K+-selective leakage current which appeared after cell perfusion with internal solutions containing different [Na+]. The concentration for half-maximal activation of KNa channel current was 39 mM and the Hill coefficient was 3.5. At [Na+]i above 12 mM, KNa channel current dominated the unspecific leakage current. The ratio of maximum KNa channel current to unspecific leakage current was 45. 3. KNa channel current was not activated by internal Li+. It was suppressed by external 20 mM Cs+ but not by 10 mM tetraethylammonium. 4. Single KNa channels with a conductance of 142 pS in 155 mM external K+ (K+o)-85 mM internal K+ (K+i) solutions were observed at a high density of about 2 channels micrometer-2. 5. In two-electrode experiments, a direct correlation was seen between development of whole- cell KNa channel current and activation of single KNa channels during perfusion of the neurone with Na+-containing internal solution. 6. Under current-clamp conditions, KNa channels did not contribute to the action potential. However, internal perfusion of the neurone with Na+ shifted the resting potential towards the equilibrium potential for K+ (EK). Varying external [K+] indicated that in neurones perfused with Na+-containing internal solution the resting potential followed the EK values predicted by the Nernst equation over a broader voltage range than in neurones perfused with Na+-free solution. 7. It is concluded that the function of KNa channels has no links to firing behaviour but that the channels could be involved in setting or stabilizing the resting potential in small DRG neurones.
摘要
采用膜片钳技术,在大鼠背根神经节切片中直径较小(20 - 25微米)的背根神经节(DRG)神经元上研究了全细胞钠激活钾(KNa)通道电流和单个KNa通道。2. 全细胞KNa通道电流被确定为一种额外的钾选择性漏电流,在用含有不同[Na⁺]的内部溶液灌注细胞后出现。KNa通道电流的半数最大激活浓度为39 mM,希尔系数为3.5。当[Na⁺]i高于12 mM时,KNa通道电流主导非特异性漏电流。最大KNa通道电流与非特异性漏电流的比值为45。3. KNa通道电流不被内部Li⁺激活。它被外部20 mM Cs⁺抑制,但不被10 mM四乙铵抑制。4. 在155 mM外部钾(K⁺o) - [原文此处有误,应为85 mM外部钠(Na⁺o)] - 85 mM内部钾(K⁺i)溶液中,观察到单个KNa通道的电导为142 pS,且通道密度较高,约为2个通道/微米²。5. 在两电极实验中,在用含钠内部溶液灌注神经元的过程中,全细胞KNa通道电流的发展与单个KNa通道的激活之间存在直接相关性。6. 在电流钳条件下,KNa通道对动作电位没有贡献。然而,用Na⁺对神经元进行内部灌注会使静息电位向钾平衡电位(EK)移动。改变外部[K⁺]表明,在用含钠内部溶液灌注的神经元中,静息电位在比用无钠溶液灌注的神经元更宽的电压范围内遵循能斯特方程预测的EK值。7. 得出的结论是,KNa通道的功能与放电行为无关,但这些通道可能参与设定或稳定小DRG神经元的静息电位。 [注:原文中“155 mM external K+ (K+o)-85 mM internal K+ (K+i)”存在错误,推测应为“155 mM external K+ (K+o)-85 mM external Na+ (Na+o)”,翻译时已按推测修改内容进行翻译,若实际情况并非如此,请根据正确内容调整译文。]