Botany School, University of Cambridge, Downing Street, Cambridge CB2 3EA, United Kingdom.
Plant Physiol. 1986 Oct;82(2):417-22. doi: 10.1104/pp.82.2.417.
The electrophysiological properties of cytoplasm-rich fragments (single membrane samples) prepared from internodal cells of Chara corallina were explored in conjunction with K(+)-sensitive microelectrode and current-voltage (I-V) measurements. This system eliminated the problem of the inaccessible cytoplasmic layer, while preserving many of the electrical characteristics of the intact cells. In 0.1 millimolar external K concentration (K(o) (+)), the resting conductance (membrane conductance G(m), 0.85 +/- 0.25 Siemens per square meter (+/-standard error)) of the single membrane samples, was dominated by the proton pump, as suggested by the response of the near-linear I-V characteristic to changes in external pH. Initial cytoplasmic K(+) activities (a(K+)), judged most reliable, gave values of 117 +/- 67 millimolar; stable a(K+) values were 77 +/- 31 millimolar. Equilibrium potentials for K(+) (Nernst equilibrium potential) (E(K)) calculated, using either of these data sets, were near the mean membrane potential (V(m)). On a cell-to-cell basis, however, E(K) was generally negative of the V(m), despite an electrogenic contribution from the Chara proton pump. When K(o) (+) was increased to 1.0 millimolar or above, G(m) rose (by 8- to 10-fold in 10 millimolar K(o) (+)), the steady state I-V characteristics showed a region of negative slope conductance, and V(m) followed E(K). These results confirm previous studies which implicated a K(o) (+)-induced and voltage-dependent permeability to K(+) at the Chara plasma membrane. They provide an explanation for transitions between apparent K(o) (+)-insensitive and K(o) (+)-sensitive (;K(+) electrode') behavior displayed by the membrane potential, as recorded in many algae and higher plant cells.
从珊瑚藻的节间细胞中制备富含细胞质的片段(单膜样品),并结合 K(+)敏感微电极和电流-电压(I-V)测量来探索其电生理特性。该系统解决了细胞质层不可接近的问题,同时保留了完整细胞的许多电特性。在 0.1 毫摩尔外部 K 浓度(K(o) (+))下,单膜样品的静息电导(膜电导 G(m),0.85 +/- 0.25 西门子/平方米(+/-标准误差))主要由质子泵决定,这反映在近线性 I-V 特性对外部 pH 变化的响应中。初始细胞质 K(+)活度(a(K+)),判断最可靠,给出 117 +/- 67 毫摩尔的值;稳定的 a(K+)值为 77 +/- 31 毫摩尔。使用这两个数据集之一计算的 K(+)平衡电位(Nernst 平衡电位)(E(K))接近平均膜电位(V(m)). 然而,在细胞间基础上,E(K)通常为负于 V(m),尽管有 Chara 质子泵的电生成贡献。当 K(o) (+)增加到 1.0 毫摩尔或更高时,G(m)上升(在 10 毫摩尔 K(o) (+)中增加 8-10 倍),稳态 I-V 特性显示出负斜率电导区,并且 V(m)跟随 E(K)。这些结果证实了先前的研究,这些研究表明 Chara 质膜存在 K(o) (+)诱导和电压依赖性对 K(+)的通透性。它们解释了在许多藻类和高等植物细胞中记录的膜电位表现出的明显对 K(o) (+)不敏感和对 K(o) (+)敏感(“K(+)电极')行为之间的转变。