Reddy M M, Riordan J R, Quinton P M
Division of Biomedical Sciences, University of California, Riverside 92521-0121.
In Vitro Cell Dev Biol. 1988 Sep;24(9):905-10. doi: 10.1007/BF02623901.
The present investigation was undertaken to examine the usefulness of cultured human sweat duct cells for ion transport and related studies in the genetic disease, cystic fibrosis. Electrical properties of cultured duct (CD) cells were compared with electrical properties of microperfused duct (MPD) cells. The resting apical membrane potential (Va) of the CD cells was -26.4 +/- 0.9 mV, n = 158 cells as compared to -24.3 +/- 0.6 mV, n = 105 of MPD cells. The Na+-K+ pump inhibitor ouabain, when applied to the apical surface of the CD cells and basolateral surface of MPD cells, depolarized both CD cells (from -28.6 +/- 3.6 to -16.8 +/- 2.4 mV, n = 5) and MPD cells (from -23.8 +/- 0.5 mV to -19.5 +/- 1.8 mV, n = 6). The Na+ conductance inhibitor amiloride applied to the apical surface hyperpolarized the apical membrane potentials (Va) of CD cells and MPD cells by -13.2 +/- 1.4 mV, n = 43 and -34.3 +/- 3.1 mV, n = 19), respectively, indicating the presence of amiloride sensitive Na+ channels in both groups of cells. However, the amiloride sensitivity of CD cells was dependent on the age of the culture. Cl- substitution at the apical side by the impermeant anion gluconate depolarized the Va of CD cells and MPD cells by 12.2 +/- 0.9 mV, n = 32 and 37.9 +/- 4.3 mV, n = 12, respectively. The effect of beta-adrenergic agonist isoproterenol (IPR), was inconsistent. In CD cells, IPR either hyperpolarized (delta Va = -8.3 +/- 1.2 mV, n = 5) or depolarized (delta Va = 8.2 +/- 2.3 mV, n = 4) or had no effect, n = 2. In contrast, most of the MPD cells did not respond to IPR, but three cells had a varied response to IPR. Our results suggest that CD cells, like MPD cells, retain significant Na+ and Cl- conductances. CD cells seem to have developed a higher sensitivity to beta-adrenergic stimulation in tissue culture as compared to MPD cells.
本研究旨在探讨培养的人汗腺导管细胞在遗传性疾病囊性纤维化的离子转运及相关研究中的作用。将培养的导管（CD）细胞的电特性与微灌流导管（MPD）细胞的电特性进行比较。CD细胞的静息顶端膜电位（Va）为-26.4±0.9 mV，n = 158个细胞，而MPD细胞的静息顶端膜电位为-24.3±0.6 mV，n = 105个细胞。将Na + -K +泵抑制剂哇巴因分别应用于CD细胞的顶端表面和MPD细胞的基底外侧表面时，CD细胞（从-28.6±3.6 mV变为-16.8±2.4 mV，n = 5）和MPD细胞（从-23.8±0.5 mV变为-19.5±1.8 mV，n = 6）均发生去极化。将Na +电导抑制剂氨氯吡脒应用于顶端表面时，CD细胞和MPD细胞的顶端膜电位（Va）分别超极化-13.2±1.4 mV，n = 43和-34.3±3.1 mV，n = 19，表明两组细胞中均存在氨氯吡脒敏感的Na +通道。然而，CD细胞对氨氯吡脒的敏感性取决于培养物的年龄。用不透性阴离子葡萄糖酸盐在顶端侧替代Cl - 时，CD细胞和MPD细胞的Va分别去极化12.2±0.9 mV，n = 32和37.9±4.3 mV，n = 12。β-肾上腺素能激动剂异丙肾上腺素（IPR）的作用不一致。在CD细胞中，IPR要么使Va超极化（ΔVa = -8.3±1.2 mV，n = 5），要么使Va去极化（ΔVa = 8.2±2.3 mV，n = 4），要么无作用（n = 2）。相比之下，大多数MPD细胞对IPR无反应，但有三个细胞对IPR有不同反应。我们的结果表明，CD细胞与MPD细胞一样，保留了显著的Na +和Cl - 电导。与MPD细胞相比，CD细胞在组织培养中似乎对β-肾上腺素能刺激产生了更高的敏感性。
