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Variable ratio of permeability to gating charge of rBIIA sodium channels and sodium influx in Xenopus oocytes.

机译:非洲爪蟾卵母细胞中rBIIA钠通道和钠流入的通透性与门控电荷的可变比率。

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摘要

Whole-cell gating current recording from rat brain IIA sodium channels in Xenopus oocytes was achieved using a high-expression system and a newly developed high-speed two-electrode voltage-clamp. The resulting ionic currents were increased by an order of magnitude. Surprisingly, the measured corresponding gating currents were approximately 5-10 times larger than expected from ionic permeability. This prompted us to minimize uncertainties about clamp asymmetries and to quantify the ratio of sodium permeability to gating charge, which initially would be expected to be constant for a homogeneous channel population. The systematic study, however, showed a 10- to 20-fold variation of this ratio in different experiments, and even in the same cell during an experiment. The ratio of P(Na)/Q was found to correlate with substantial changes observed for the sodium reversal potential. The data suggest that a cytoplasmic sodium load in Xenopus oocytes or the energy consumption required to regulate the increase in cytoplasmic sodium represents a condition where most of the expressed sodium channels keep their pore closed due to yet unknown mechanisms. In contrast, the movements of the voltage sensors remain undisturbed, producing gating current with normal properties.
机译:使用高表达系统和新开发的高速两电极电压钳实现了非洲爪蟾卵母细胞中大鼠脑IIA钠通道的全细胞门控电流记录。产生的离子电流增加了一个数量级。令人惊讶的是,测得的相应门控电流比离子渗透率预期的大约5-10倍。这促使我们将关于钳位不对称性的不确定性减至最小,并量化钠渗透率与门控电荷的比率,该比率最初预计对于均匀通道总数是恒定的。然而,系统的研究表明,在不同的实验中,甚至在实验过程中,在同一细胞中,该比例的变化为10到20倍。发现P(Na)/ Q的比例与观察到的钠反转电位的实质性变化相关。数据表明,爪蟾卵母细胞中的细胞质钠负荷或调节细胞质钠增加所需的能量消耗代表了一种情况,其中大多数表达的钠通道由于未知的机制而使其孔保持封闭。相反,电压传感器的运动保持不受干扰,产生具有正常特性的门控电流。

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