Timbs M M, Spring K R
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892-1598, USA.
J Membr Biol. 1996 Sep;153(1):1-11. doi: 10.1007/s002329900104.
The water permeability of the apical and basolateral cell membranes and the compliance of the lateral intercellular spaces (LIS) of MDCK monolayers were measured on confluent cultures grown on permeable supports. Cell membrane water permeabilities were determined, using quantitative differential interference light microscopy, from the rate of cell volume decrease after exposure to a hyperosmotic bathing solution. Both membranes exhibited osmotic water permeabilities (POSM) of approximately 10 microm/sec, comparable to that of unmodified lipid bilayers. The compliance of the cell membranes forming the lateral intercellular space (LIS) between cells was determined from the pressure-volume relation. Confocal microscopy of fluorescent labeling of the basolateral cell membranes was used to delineate the LIS geometry as transepithelial hydrostatic pressure was varied. The LIS were poorly deformable as a function of transepithelial hydrostatic pressure until a pressure of >/=8 cm H2O (basolateral > apical) was reached where catastrophic failure of intercellular connections occurred. The compliance of the LIS was calculated from the geometry changes at pressures <8 cm H2O and ranged from 0.05-0.11 cm H2O-1, comparable to that previously predicted in mathematical models of the rat proximal tubule.
在可渗透支持物上生长的汇合培养物中,测量了MDCK单层细胞顶膜和基底外侧细胞膜的水渗透性以及细胞间侧间隙(LIS)的顺应性。使用定量微分干涉光学显微镜,通过暴露于高渗浴液后细胞体积减小的速率来测定细胞膜的水渗透性。两种膜的渗透水渗透率(POSM)约为10微米/秒,与未修饰的脂质双层相当。根据压力-体积关系确定形成细胞间侧间隙(LIS)的细胞膜的顺应性。当跨上皮静水压力变化时,使用基底外侧细胞膜荧光标记的共聚焦显微镜来描绘LIS的几何形状。直到达到> /= 8 cm H2O(基底外侧>顶侧)的压力,细胞间连接发生灾难性破坏之前,LIS作为跨上皮静水压力的函数难以变形。LIS的顺应性是根据压力<8 cm H2O时的几何形状变化计算得出的,范围为0.05 - 0.11 cm H2O-1,与先前在大鼠近端小管数学模型中预测的结果相当。