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22、10 和 0°C 时 COS-7 细胞水通透性的校正。

Rectification of the water permeability in COS-7 cells at 22, 10 and 0°C.

机构信息

Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.

出版信息

PLoS One. 2011;6(8):e23643. doi: 10.1371/journal.pone.0023643. Epub 2011 Aug 24.

Abstract

The osmotic and permeability parameters of a cell membrane are essential physico-chemical properties of a cell and particularly important with respect to cell volume changes and the regulation thereof. Here, we report the hydraulic conductivity, L(p), the non-osmotic volume, V(b), and the Arrhenius activation energy, E(a), of mammalian COS-7 cells. The ratio of V(b) to the isotonic cell volume, V(c iso), was 0.29. E(a), the activation energy required for the permeation of water through the cell membrane, was 10,700, and 12,000 cal/mol under hyper- and hypotonic conditions, respectively. Average values for L(p) were calculated from swell/shrink curves by using an integrated equation for L(p). The curves represented the volume changes of 358 individually measured cells, placed into solutions of nonpermeating solutes of 157 or 602 mOsm/kg (at 0, 10 or 22°C) and imaged over time. L(p) estimates for all six combinations of osmolality and temperature were calculated, resulting in values of 0.11, 0.21, and 0.10 µm/min/atm for exosmotic flow and 0.79, 1.73 and 1.87 µm/min/atm for endosmotic flow (at 0, 10 and 22°C, respectively). The unexpected finding of several fold higher L(p) values for endosmotic flow indicates highly asymmetric membrane permeability for water in COS-7. This phenomenon is known as rectification and has mainly been reported for plant cell, but only rarely for animal cells. Although the mechanism underlying the strong rectification found in COS-7 cells is yet unknown, it is a phenomenon of biological interest and has important practical consequences, for instance, in the development of optimal cryopreservation.

摘要

细胞膜的渗透和通透性参数是细胞的重要物理化学性质,特别是对于细胞体积变化及其调节具有重要意义。在这里,我们报告了哺乳动物 COS-7 细胞的水力传导率 L(p)、非渗透体积 V(b)和阿仑尼乌斯活化能 E(a)。V(b)与等渗细胞体积 V(c iso)的比值为 0.29。E(a)是水通过细胞膜渗透所需的活化能,在高渗和低渗条件下分别为 10700 和 12000 千卡/摩尔。L(p)的平均值是根据膨胀/收缩曲线通过使用 L(p)的积分方程计算得出的。这些曲线代表了 358 个单独测量的细胞的体积变化,这些细胞被放置在非渗透溶质的溶液中(渗透压为 157 或 602 mOsm/kg,温度为 0、10 或 22°C),并随时间成像。对所有六种渗透压和温度组合的 L(p)估计进行了计算,结果得到外渗流的 0.11、0.21 和 0.10 µm/min/atm,内渗流的 0.79、1.73 和 1.87 µm/min/atm(分别在 0、10 和 22°C)。内渗流的 L(p)值高出数倍的意外发现表明 COS-7 中对水的膜通透性具有高度不对称性。这种现象称为整流,主要在植物细胞中报道,但在动物细胞中很少报道。尽管在 COS-7 细胞中发现的强烈整流的机制尚不清楚,但它是一种具有生物学意义的现象,对优化冷冻保存的发展具有重要的实际意义。

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