The interaction between K+, Cl- and H2O fluxes was studied in the ventricular membrane of the choroid plexus epithelium from Necturus maculosus by means of ion-selective microelectrodes. 2. Three experimental strategies were adopted: the osmolarity of the ventricular solution was increased abruptly by addition of (i) mannitol or (ii) KCl; (iii) Na+ in the ventricular solution was replaced isosmotically by K+. 3. The mannitol experiments showed that H2O had two pathways across the ventricular membrane. One was purely passive, with a water permeability, L'p, of 0.64 x 10(-4) cm s-1 (osmol l-1)-1. This operated in parallel with an ion-dependent pathway of similar magnitude which was abolished in Cl(-)-free solutions. 4. When KCl was added there was a flow of H2O into the cell. Surprisingly, this took place despite the osmotic gradient which favoured an efflux of H2O. The effect was blocked by frusemide (furosemide), in which case KCl had the same effects as applications of NaCl or mannitol. 5. Replacement of Na+ with K+ caused an influx of H2O. This flux could proceed against osmotic gradients implemented by mannitol. 6. The present data and those of earlier publications show that the interdependence of the fluxes of K+, Cl- and H2O in the exit membrane can be described as cotransport. The fluxes have a fixed stoichiometry of 1:1:500, the flux of one species is able to energize the flux of the two others, and the transport exhibits saturation and is specific for K+ and Cl-. 7. A molecular model based upon a mobile barrier in a membrane spanning protein gives an accurate quantitative description of the data.
摘要
采用离子选择性微电极研究了黄斑美西螈脉络丛上皮心室膜中钾离子(K⁺)、氯离子(Cl⁻)和水通量之间的相互作用。2. 采用了三种实验策略:通过添加(i)甘露醇或(ii)氯化钾突然增加心室溶液的渗透压;(iii)用钾离子等渗替代心室溶液中的钠离子。3. 甘露醇实验表明,水有两条穿过心室膜的途径。一条是纯粹被动的,水渗透率L'p为0.64×10⁻⁴ cm s⁻¹(渗透压单位⁻¹)。这条途径与一条大小相似的离子依赖性途径并行运作,在无氯离子的溶液中该离子依赖性途径被消除。4. 添加氯化钾时,水流入细胞。令人惊讶的是,尽管渗透压梯度有利于水外流,但这种情况仍会发生。该效应被速尿(呋塞米)阻断,在这种情况下,氯化钾与应用氯化钠或甘露醇具有相同的效果。5. 用钾离子替代钠离子导致水的流入。这种通量可以逆着由甘露醇形成的渗透压梯度进行。6. 目前的数据和早期出版物的数据表明,出口膜中钾离子、氯离子和水通量的相互依赖性可以描述为协同转运。通量具有固定的化学计量比1:1:500,一种物质的通量能够为其他两种物质的通量提供能量,并且这种转运表现出饱和性,且对钾离子和氯离子具有特异性。7. 基于跨膜蛋白中移动屏障的分子模型对数据给出了准确的定量描述。