Rettinger J
Max-Planck-Institut für Biophysik/JWG-University Frankfurt, Kennedyallee 70, D-60596 Frankfurt/Main, Germany.
Pflugers Arch. 1999 May;437(6):917-24. doi: 10.1007/s004240050863.
It has been shown by means of the two-microelectrode voltage-clamp technique that in membranes of Xenopus laevis oocytes a Na+-selective permeability can be activated by long-lasting or repetitive depolarization (R.T. Kado and C. Baud, Journal of Physiology, Paris, 77:1113-1117, 1981). In this study the permeability in inside-out giant membrane patches with diameters of 20-30 microm was analysed. Once induced, the Na+ permeability has a voltage-dependent open probability that increases with positive potentials and half-maximally activates at about 0 mV. Sudden changes of membrane potential elicit transient currents with strongly voltage-dependent time constants of from less than 1 ms at -150 mV to several hundreds of milliseconds at positive potentials. In contrast to the on-cell configuration, the permeability ceases completely within a few minutes in the cell-free inside-out configuration. This rundown can be prevented by including MgATP, but not Mg2+ or ATP alone, in the intracellular solution. Intracellular Mg2+ ions, in addition to being a co-factor for ATP in the activation process, decrease the permeability in a dose-dependent manner. Steady-state fluctuation analysis gave no evidence that an increased noise level is caused by open-close kinetics of an ion channel, suggesting that the single-channel conductance is below 1 pS if a channel-like structure is the origin of the endogenous Na+ permeability.
运用双微电极电压钳技术已表明,在非洲爪蟾卵母细胞的膜中,持久或重复去极化可激活钠选择性通透性(R.T. 卡多和C. 鲍德,《巴黎生理学杂志》,77:1113 - 1117,1981年)。在本研究中,分析了直径为20 - 30微米的内向外巨膜片的通透性。一旦被诱导,钠通透性具有电压依赖性开放概率,该概率随正电位增加,并在约0 mV时半最大激活。膜电位的突然变化引发瞬态电流,其时间常数强烈依赖电压,从 - 150 mV时小于1毫秒到正电位时几百毫秒不等。与在细胞膜上的配置不同,在无细胞的内向外配置中,通透性在几分钟内完全停止。通过在细胞内溶液中加入MgATP而非单独的Mg²⁺或ATP,可防止这种衰减。细胞内的Mg²⁺离子,除了在激活过程中作为ATP的辅助因子外,还以剂量依赖方式降低通透性。稳态波动分析没有证据表明离子通道的开闭动力学导致噪声水平增加,这表明如果类似通道的结构是内源性钠通透性的起源,则单通道电导低于1 pS。