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Membrane chloride conductance and capacitance in Jurkat T lymphocytes during osmotic swelling.

机译:渗透溶胀过程中Jurkat T淋巴细胞的膜氯化物电导和电容。

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摘要

Video microscopy and whole-cell patch-clamp recording were used to monitor changes in relative cell volume (V/Vo), chloride conductance (gCl), and membrane capacitance (Cm) during osmotically induced swelling in Jurkat T lymphocytes. Cellular swelling was initiated with hyperosmotic pipette solutions. Simultaneous evaluation of V/Vo and gCl revealed a 59-s delay between the inception of swelling and the activation of outwardly rectifying, ATP-dependent Cl- channels. Following the delay, increases in V/Vo and gCl progressed in parallel. In contrast, Cm, a measure of cell surface area, fell gradually at a rate of approximately 150 fF/min after whole-cell access was achieved. The decline in Cm lasted 200 s and was followed by a rapid rise (approximately 750 fF/min). The rise in Cm coincided with a variable increase in "leak" current, gCl increased at a slower rate and reached lower peak values in experiments performed without ATP; ATP had no effect on the biphasic Cm time course. The temporal separation of conductance and capacitance during swelling suggests that gCl and Cm vary independently, supporting the hypothesis that a large portion, if not all, of the whole-cell Cl- conductance activated during swelling is provided by volume-sensitive Cl- channels preexisting in the plasma membrane.
机译:使用视频显微镜和全细胞膜片钳记录来监测渗透压引起的Jurkat T淋巴细胞肿胀期间相对细胞体积(V / Vo),氯化物电导(gCl)和膜电容(Cm)的变化。用高渗移液器溶液开始细胞肿胀。对V / Vo和gCl的同时评估显示,溶胀开始与向外整流的ATP依赖性Cl-通道激活之间存在59秒的延迟。在延迟之后,V / Vo和gCl平行增加。相比之下,Cm,一种细胞表面积的测量,在达到全细胞通路后以约150 fF / min的速率逐渐下降。 Cm的下降持续了200 s,然后迅速上升(大约750 fF / min)。在没有ATP的实验中,Cm的上升与“泄漏”电流的变化不一而足,gCl的上升速度较慢,并且达到较低的峰值; ATP对双相Cm时间进程没有影响。溶胀过程中电导和电容的时间分离表明gCl和Cm独立变化,支持以下假设:溶胀过程中激活的全细胞Cl-电导的很大一部分(如果不是全部)由预先存在的体积敏感Cl-通道提供在质膜上

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