Persson B E, Spring K R
J Gen Physiol. 1982 Mar;79(3):481-505. doi: 10.1085/jgp.79.3.481.
The hydraulic water permeability (Lp) of the cell membranes of Necturus gallbladder epithelial cells was estimated from the rate of change of cell volume after a change in the osmolality of the bathing solution. Cell volume was calculated from computer reconstruction of light microscopic images of epithelial cells obtained by the "optical slice" technique. The tissue was mounted in a miniature Ussing chamber designed to achieve optimal optical properties, rapid bath exchange, and negligible unstirred layer thickness. The control solution contained only 80% of the normal NaCl concentration, the remainder of the osmolality was made up by mannitol, a condition that did not significantly decrease the fluid absorption rate in gallbladder sac preparations. The osmotic gradient ranged from 11.5 to 41 mosmol and was achieved by the addition or removal of mannitol from the perfusion solutions. The Lp of the apical membrane of the cell was 1.0 X 10(-3) cm/s . osmol (Posm = 0.055 cm/s) and that of the basolateral membrane was 2.2 X 10(-3) cm/s . osmol (Posm = 0.12 cm/s). These values were sufficiently high so that normal fluid absorption by Necturus gallbladder could be accomplished by a 2.4-mosmol solute gradient across the apical membrane and a 1.1-mosmol gradient across the basolateral membrane. After the initial cell shrinkage or swelling resulting from the anisotonic mucosal or serosal medium, cell volume returned rapidly toward the control value despite the fact that one bathing solution remained anisotonic. This volume regulatory response was not influenced by serosal ouabain or reduction of bath NaCl concentration to 10 mM. Complete removal of mucosal perfusate NaCl abolished volume regulation after cell shrinkage. Estimates were also made of the reflection coefficient for NaCl and urea at the apical cell membrane and of the velocity of water flow across the cytoplasm.
通过改变浴液渗透压后细胞体积的变化率来估算美西螈胆囊上皮细胞膜的水力水渗透率(Lp)。细胞体积是根据通过“光学切片”技术获得的上皮细胞光学显微镜图像的计算机重建来计算的。将组织安装在一个微型Ussing室中,该室设计用于实现最佳光学性能、快速的浴液交换以及可忽略不计的未搅拌层厚度。对照溶液仅含有正常NaCl浓度的80%,渗透压的其余部分由甘露醇补足,这种情况不会显著降低胆囊囊泡制剂中的液体吸收率。渗透压梯度范围为11.5至41毫渗摩尔,通过向灌注溶液中添加或去除甘露醇来实现。细胞顶端膜的Lp为1.0×10⁻³厘米/秒·渗透压(Posm = 0.055厘米/秒),基底外侧膜的Lp为2.2×10⁻³厘米/秒·渗透压(Posm = 0.12厘米/秒)。这些值足够高,以至于美西螈胆囊正常的液体吸收可以通过跨顶端膜2.4毫渗摩尔的溶质梯度和跨基底外侧膜1.1毫渗摩尔的梯度来完成。在由等渗的黏膜或浆膜介质导致最初的细胞收缩或肿胀后,尽管一种浴液仍为等渗,但细胞体积迅速恢复到对照值。这种体积调节反应不受浆膜哇巴因或将浴液NaCl浓度降低至10 mM的影响。完全去除黏膜灌注液中的NaCl会消除细胞收缩后的体积调节。还对顶端细胞膜处NaCl和尿素的反射系数以及跨细胞质的水流速度进行了估算。