York University, Department of Biology, 4700 Keele Street, North York, Ontario M3J 1P3, Canada.
Plant Physiol. 1991 Dec;97(4):1527-34. doi: 10.1104/pp.97.4.1527.
Ion transport, measured using double-barreled micropipettes to obtain current-voltage relations, was examined in Arabidopsis thaliana root hairs that continued tip growth and cytoplasmic streaming after impalement with the micropipette. To do this required in situ measurements with no handling of the seedlings to avoid wounding responses, and conditions allowing good resolution microscopy in tandem with the electrophysiological measurements. Two ion transport processes were demonstrated. One was a tetraethylammonium-sensitive potassium ion current, inward at hyperpolarized potentials and outward at depolarized potentials. The addition of tetraethylammonium (a potassium channel blocker) caused the potential to hyperpolarize, indicating the presence of a net inward potassium current through the ion channels at the resting potential. The potassium influx was sufficient to "drive" cellular expansion based upon growth rates. Indeed, tetraethylammonium caused transient inhibition of tip growth. The other electrogenic process was the plasma membrane proton pump, measured by indirect inhibition with cyanide or direct inhibition by vanadate. The proton pump was the dominant contribution to the resting potential, with a very high current density of about 250 microamperes per square centimeter (seen only in young growing root hairs). The membrane potential generated by the proton pump presumably drives the potassium influx required for cellular expansion. The pump appears to be a constant current source over the voltage range -200 to 0 millivolts. With this system, it is now possible to study the physiology of a higher plant cell in dynamic living state using a broad range of cell biological and electrophysiological techniques.
离子转运的研究采用双管微电极来获得电流-电压关系,对完整的拟南芥根毛细胞进行研究,这些根毛在刺穿微电极后仍能进行尖端生长和细胞质流动。要实现这一目标,需要在不处理幼苗的情况下进行原位测量,以避免伤害反应,并需要允许良好分辨率显微镜与电生理测量相结合的条件。研究证明了两种离子转运过程。一种是四乙铵敏感的钾离子电流,在超极化电位下内向,在去极化电位下外向。加入四乙铵(钾通道阻断剂)会使电位超极化,表明在静息电位下,离子通道存在净内向钾电流。钾离子内流足以根据生长速率“驱动”细胞扩张。事实上,四乙铵会导致尖端生长的短暂抑制。另一种电致离子过程是质膜质子泵,通过氰化物间接抑制或钒酸盐直接抑制来测量。质子泵是静息电位的主要贡献者,其电流密度非常高,约为 250 微安培/平方厘米(仅见于年轻的生长根毛中)。质子泵产生的膜电位可能驱动细胞扩张所需的钾离子内流。该泵在-200 至 0 毫伏的电压范围内似乎是一个恒流源。有了这个系统,现在就可以使用广泛的细胞生物学和电生理学技术来研究动态活体高等植物细胞的生理学。