Department of Physiology, Medical School, University Walk, Bristol BS8 1TD, UK.
J Exp Biol. 1989 Sep;145:455-64. doi: 10.1242/jeb.145.1.455.
The effects of external Zn(2+) on Ca(2+) and H+ currents in the soma of intact Helix neurones were investigated using standard two-electrode voltage-clamp procedures. Cells were exposed to a 0Na(+), tetraethylammonium (TEA(+)) saline and clamped with Cs(+)-filled electrodes, which allows separation of voltage-dependent H(+) and Ca(2+) currents using different holding potentials. Outward H(+) currents, activated by depolarizations from holding potentials in the range -15 to-10 mV, were rapidly blocked by low concentrations of external Zn(2+) with a K(d) of approximately 16/μmoll(-1). H(+) current activation was also markedly slowed and the block was slow to reverse. Ca(2+) currents, largely free from contamination by outward current, were activated by small depolarizations from a holding potential of -55 mV. Ca(2+) currents were reduced by Zn(2+), but the K(d) for block was more than 80 times greater than for block of H(+) currents. Thus, low concentrations of Zn(2+) provide a method of selectively inhibiting H(+) current in studies of Ca(2+) current. This was demonstrated in cells which slowly acidified following exposure to 0Na(+), TEA(+) saline, leading to an increased outward H(+) current. Washing with low concentrations of Zn(2+) blocked the H(+) current and uncovered the underlying Ca(2+) current. The results also suggest that Zn(2+) will be a useful tool in studies of the physiological role of the H(+) pathway.
采用标准的双电极电压钳技术研究了外源性 Zn(2+) 对完整海兔神经元体 Ca(2+) 和 H+电流的影响。细胞暴露于 0Na(+)、四乙基铵 (TEA(+)) 盐溶液中,并使用 Cs(+) 填充的电极进行钳制,这允许使用不同的保持电位分离电压依赖性 H(+)和 Ca(2+)电流。从-15 到-10 mV 的保持电位去极化激活外向 H(+)电流,低浓度的外源性 Zn(2+) 可迅速阻断该电流,K(d) 值约为 16/μmoll(-1)。H(+)电流的激活也明显减慢,阻断的恢复也很慢。Ca(2+)电流,很大程度上不受外向电流的污染,由从-55 mV 的保持电位的小去极化激活。Zn(2+) 减少 Ca(2+)电流,但阻断的 K(d) 值比阻断 H(+)电流的值大 80 多倍。因此,低浓度的 Zn(2+) 为研究 Ca(2+)电流时选择性抑制 H(+)电流提供了一种方法。这在暴露于 0Na(+)、TEA(+)盐溶液后缓慢酸化的细胞中得到了证明,这导致外向 H(+)电流增加。用低浓度的 Zn(2+) 洗涤可阻断 H(+)电流并揭示潜在的 Ca(2+)电流。结果还表明,Zn(2+) 将成为研究 H(+)途径生理作用的有用工具。