Fein Alan, Terasaki Mark
Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
J Gen Physiol. 2005 Aug;126(2):151-9. doi: 10.1085/jgp.200509294.
Plasma membrane wound repair is an important but poorly understood process. We used femtosecond pulses from a Ti-Sapphire laser to make multiphoton excitation-induced disruptions of the plasma membrane while monitoring the membrane potential and resistance. We observed two types of wounds that depolarized the plasma membrane. At threshold light levels, the membrane potential and resistance returned to prewound values within seconds; these wounds were not easily observed by light microscopy and resealed in the absence of extracellular Ca(2+). Higher light intensities create wounds that are easily visible by light microscopy and require extracellular Ca(2+) to reseal. Within a few seconds the membrane resistance is approximately 100-fold lower, while the membrane potential has depolarized from -80 to -30 mV and is now sensitive to the Cl(-) concentration but not to that of Na(+), K(+), or H(+). We suggest that the chloride sensitivity of the membrane potential, after wound resealing, is due to the fusion of chloride-permeable intracellular membranes with the plasma membrane.
质膜伤口修复是一个重要但却知之甚少的过程。我们使用钛宝石激光器发出的飞秒脉冲,在监测膜电位和电阻的同时,使质膜发生多光子激发诱导的破坏。我们观察到两种使质膜去极化的伤口类型。在阈值光照水平下,膜电位和电阻在数秒内恢复到受伤前的值;这些伤口在光学显微镜下不易观察到,并且在没有细胞外Ca(2+)的情况下重新封闭。更高的光照强度会造成在光学显微镜下很容易看到的伤口,并且需要细胞外Ca(2+)来重新封闭。在几秒钟内,膜电阻大约降低100倍,而膜电位则从-80 mV去极化到-30 mV,并且现在对Cl(-)浓度敏感,而对Na(+)、K(+)或H(+)的浓度不敏感。我们认为,伤口重新封闭后膜电位对氯离子的敏感性,是由于氯离子可通透的细胞内膜与质膜融合所致。