Cameron I L, Hardman W E, Fullerton G D, Miseta A, Koszegi T, Ludany A, Kellermayer M
Department of Cellular and Structural Biology, University of Texas Health Science Center at San Antonio 78284, USA.
Cell Biol Int. 1996 Feb;20(2):127-37. doi: 10.1006/cbir.1996.0017.
If the plasma membrane and its associated transport proteins are solely responsible for maintenance of the asymmetric solute distribution then disruption of the plasma membrane would quickly lead to the symmetric distribution of all unattached inorganic ions between the cell and the extracellular environment. To test this hypothesis fresh pig lenses were incubated in Hanks' balanced salt solution in either absence or presence of non-ionic detergents (0.2% Triton X-100 or 0.2% Brij 58). Both detergents caused permeabilization of every lens fiber cell as shown by electron microscopy. The flux kinetics of K+, Mg2+, Na+, Ca2+, water and protein out of and into the permeabilized lens fiber cells was measured. Triton X-100 caused a faster flux rate of all solutes than did Brij 58. The Triton X-100 induced flux of solutes and water was associated with a decrease in lens ATP. Incubation of untreated lenses in solutions of different osmotic pressures at 0 degree C demonstrated that the major fraction of lens water was osmotically unresponsive. Thus the asymmetric distribution of solutes in lens fiber cells is dependent on an intact plasma membrane and on a co-operative ATP-dependent association between K+, Mg2+, water and cytomatrix proteins.
如果质膜及其相关的转运蛋白是维持溶质不对称分布的唯一原因,那么质膜的破坏将迅速导致所有未结合的无机离子在细胞和细胞外环境之间对称分布。为了验证这一假设,将新鲜猪晶状体置于汉克斯平衡盐溶液中,分别在不存在或存在非离子去污剂(0.2% Triton X-100或0.2% Brij 58)的情况下孵育。电子显微镜显示,两种去污剂均导致每个晶状体纤维细胞透化。测定了K+、Mg2+、Na+、Ca2+、水和蛋白质进出透化晶状体纤维细胞的通量动力学。Triton X-100引起的所有溶质通量速率比Brij 58更快。Triton X-100诱导的溶质和水通量与晶状体ATP的减少有关。在0℃下将未处理的晶状体置于不同渗透压的溶液中孵育表明,晶状体水的主要部分对渗透压无反应。因此,晶状体纤维细胞中溶质的不对称分布取决于完整的质膜以及K+、Mg2+、水和细胞基质蛋白之间依赖ATP的协同作用。