Verkman A S, Ives H E
Am J Physiol. 1986 Apr;250(4 Pt 2):F633-43. doi: 10.1152/ajprenal.1986.250.4.F633.
Water and nonelectrolyte permeability in basolateral membrane vesicles (BLMV) isolated from rat and rabbit renal cortex were characterized. Osmotic water (Pf) and solute (Ps) permeabilities were determined from the time course of scattered light intensity in response to osmotic gradients. In rabbit BLMV, Pf = 2.5 X 10(-2) cm/s, Purea = 1.2 X 10(-6) cm/s, and sigma urea = 0.95 (23 degrees C). Relative solute permeabilities were urea, 1; thiourea, 0.83; ethylene glycol, 9.7; glycerol, 4.6; formamide, 20; and acetamide, 12. Pf and Purea were not altered by organic mercurials, phloretin, urea, and high-affinity urea analogues. delta H (12-50 degrees C) was 10.7 and 2.5 kcal/mol for Purea and Pf, respectively; in contrast, reported delta H for Pf in rabbit brush-border membrane vesicles (BBMV) was 2 kcal/mol (T less than 33 degrees C) and 14 kcal/mol (T greater than 33 degrees C). To examine whether membrane fluidity changes were associated with this difference, fluorescence anisotropy decay was measured using 1,6-diphenyl-1,3,5-hexatriene (DPH). In both BLMV and BBMV, DPH rotation was hindered at all temperatures (5-53 degrees C). Changes in steady-state anisotropy were attributable to changes in DPH rotational freedom rather than to changes in DPH rotational rate without evidence for a definitive membrane thermotropic phase transition. These results suggest that BLMV urea transport occurs by lipid diffusion and that osmotic water transport is rapid and may be facilitated. A comparison of transepithelial Pf with BLMV and BBMV Pf is consistent with transcellular osmotic water flow in the proximal tubule.
对从大鼠和兔肾皮质分离出的基底外侧膜囊泡(BLMV)的水和非电解质通透性进行了表征。根据散射光强度随渗透梯度变化的时间进程来测定渗透水(Pf)和溶质(Ps)的通透性。在兔BLMV中,Pf = 2.5×10⁻² cm/s,P尿素 = 1.2×10⁻⁶ cm/s,且尿素的反射系数σ尿素 = 0.95(23℃)。相对溶质通透性分别为:尿素,1;硫脲,0.83;乙二醇,9.7;甘油,4.6;甲酰胺,20;以及乙酰胺,12。Pf和P尿素不受有机汞剂、根皮素、尿素及高亲和力尿素类似物的影响。在12 - 50℃范围内,P尿素和Pf的ΔH分别为10.7和2.5 kcal/mol;相比之下,据报道兔刷状缘膜囊泡(BBMV)中Pf的ΔH在T<33℃时为2 kcal/mol,在T>33℃时为14 kcal/mol。为了研究膜流动性变化是否与这种差异相关,使用1,6 - 二苯基 - 1,3,5 - 己三烯(DPH)测量了荧光各向异性衰减。在BLMV和BBMV中,在所有温度(5 - 53℃)下DPH的旋转均受到阻碍。稳态各向异性的变化归因于DPH旋转自由度的变化,而非DPH旋转速率的变化,且没有明确的膜热致相变证据。这些结果表明,BLMV中的尿素转运通过脂质扩散发生,且渗透水转运迅速且可能得到促进。将跨上皮Pf与BLMV和BBMV的Pf进行比较,与近端小管中的跨细胞渗透水流一致。