van Heeswijk M P, van Os C H
J Membr Biol. 1986;92(2):183-93. doi: 10.1007/BF01870707.
The osmotic water permeability Pf of brush border (BBM) and basolateral (BLM) membrane vesicles from rat small intestine and renal cortex was studied by means of stopped-flow spectrophotometry. Scattered light intensity was used to follow vesicular volume changes upon osmotic perturbation with hypertonic mannitol solutions. A theoretical analysis of the relationship of scattered light intensity and vesicular volume justified a simple exponential approximation of the change in scattered light intensity. The rate constants extracted from fits to an exponential function were proportional to the final medium osmolarity as predicted by theory. For intestinal membranes, computer analysis of optical responses fitted well with a single-exponential treatment. For renal membranes a double-exponential treatment was needed, implying two distinct vesicle populations. Pf values for BBM and BLM preparations of small intestine were equal and amount to 60 microns/sec. For renal preparations, Pf values amount to 600 microns/sec for the fast component, BBM as well as BLM, and to 50 (BBM) and 99 (BLM) microns/sec for the slow component. The apparent activation energy for water permeation in intestinal membranes was 13.3 +/- 0.6 and in renal membranes 1.0 +/- 0.3 kCal/mole, between 25 and 35 degrees C. The mercurial sulfhydryl reagent pCMBS inhibited completely and reversibly the high Pf value in renal brush border preparations. These observations suggest that in intestinal membranes water moves through the lipid matrix but that in renal plasma membranes water channels may be involved. From the high Pf values of renal membrane vesicles a transcellular water permeability for proximal tubules can be calculated which amounts to approximately 1 cm/sec. This value allows for an entirely transcellular route for water flow during volume reabsorption.
采用停流分光光度法研究了大鼠小肠和肾皮质刷状缘(BBM)及基底外侧(BLM)膜囊泡的渗透水通透性Pf。用散射光强度跟踪高渗甘露醇溶液渗透扰动后囊泡体积的变化。对散射光强度与囊泡体积关系的理论分析证明了散射光强度变化的简单指数近似是合理的。从指数函数拟合中提取的速率常数与理论预测的最终介质渗透压成正比。对于肠膜,光学响应的计算机分析与单指数处理拟合良好。对于肾膜,需要双指数处理,这意味着存在两种不同的囊泡群体。小肠BBM和BLM制剂的Pf值相等,为60微米/秒。对于肾制剂,快速成分(BBM和BLM)的Pf值为600微米/秒,慢速成分(BBM为50微米/秒,BLM为99微米/秒)。在25至35摄氏度之间,肠膜中水渗透的表观活化能为13.3±0.6千卡/摩尔,肾膜为1.0±0.3千卡/摩尔。汞巯基试剂对氯汞苯甲酸完全且可逆地抑制了肾刷状缘制剂中的高Pf值。这些观察结果表明,在肠膜中,水通过脂质基质移动,但在肾质膜中可能涉及水通道。根据肾膜囊泡的高Pf值,可以计算近端小管的跨细胞水通透性,约为1厘米/秒。该值允许在容量重吸收期间水完全通过跨细胞途径流动。