Saitou T, Ishikawa T, Obara K, Nakayama K
Department of Pharmacology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-City, Japan.
Pflugers Arch. 2000 Oct;440(6):858-65. doi: 10.1007/s004240000337.
Whole-cell mechanosensitive current (I(ms)) in Xenopus oocytes was studied using the two-electrode voltage-clamp technique. I(ms) was evoked by mechanically pressing the oocyte surface with a glass micropipette. The current was found to depend on the amplitude of the stimulus, showed a time-dependent decay, and turned off immediately after the stimulus was removed. The current-voltage relationship for the peak current exhibited inward and outward rectification at negative and positive potentials, respectively, while that for the sustained current exhibited only inward rectification. I(ms) was significantly suppressed by 30 microM Gd3+. One millimolar amiloride also significantly suppressed the inward I(ms) at negative potentials, but not the outward one at positive potentials. Replacing extracellular Na+ with K+ did not change the current-voltage relationship, whereas replacing extracellular Na+ with choline+ or tetraethylammonium+ significantly decreased the inward I(ms). The outward rectifier at positive potentials was abolished by replacing extracellular Cl- with gluconate-, by intracellular injection of 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA), by extracellular application of anthracene-9-carboxylic acid, and by replacing extracellular Ca2+ with Mg2+. These results suggest that mechanical stimulation activates stretch-activated cation channels and Ca2+-activated Cl- channels, the latter being secondarily activated by an increase in intracellular Ca2+ concentration by Ca2+ influx through stretch-activated cation channels.
采用双电极电压钳技术研究了非洲爪蟾卵母细胞中的全细胞膜片机械敏感性电流(I(ms))。通过用玻璃微吸管机械按压卵母细胞表面来诱发I(ms)。发现该电流取决于刺激的幅度,呈现出时间依赖性衰减,并且在刺激移除后立即关闭。峰值电流的电流-电压关系在负电位和正电位分别表现出内向和外向整流,而持续电流的电流-电压关系仅表现出内向整流。30 microM的Gd3+可显著抑制I(ms)。1毫摩尔的氨氯地平也可显著抑制负电位下的内向I(ms),但对正电位下的外向I(ms)无抑制作用。用K+替代细胞外Na+不会改变电流-电压关系,而用胆碱+或四乙铵+替代细胞外Na+会显著降低内向I(ms)。通过用葡萄糖酸盐-替代细胞外Cl-、通过细胞内注射1,2-双(2-氨基苯氧基)乙烷-N,N,N',N'-四乙酸(BAPTA)、通过细胞外应用蒽-9-羧酸以及通过用Mg2+替代细胞外Ca2+,可消除正电位下的外向整流。这些结果表明,机械刺激激活了牵张激活阳离子通道和Ca2+激活的Cl-通道,后者是由通过牵张激活阳离子通道的Ca2+内流导致细胞内Ca2+浓度升高而继发激活的。