Larsson L, Aperia A, Lechene C
Acta Physiol Scand. 1986 Mar;126(3):321-32. doi: 10.1111/j.1748-1716.1986.tb07823.x.
Renal epithelial cells were isolated from the outermost superficial cortex of adult and young rats. The cells, likely of proximal origin, were plated on silicon pieces, and cultured during 1-3 days. Intracellular content and concentrations of K, Na, Cl, and P, and the kinetics of change in intracellular content, after inhibition of Na-K ATPase by incubation with ouabain or in K-free medium, were measured in individual cells in small populations using electron probe analysis. In control medium, concentrations in mM were approximately: K, 130; Na, 15; Cl, 28; P, 140. After 6 h inhibition of Na-K ATPase, cells exchanged all K for Na, and the intracellular Na concentration increased to 139 mM in K-free medium. The Cl concentration increased at most to 46 mM. The sum of intracellular K + Na + Cl did not increase more than 25% after 24 h incubation in K-free medium. There were no differences in intracellular K, Na, and Cl for adult and young rat cells in similar conditions. The half-times of K efflux and Na influx after inhibition of Na-K ATPase measured in adult rat were approximately 16-20 min. In the absence of serum, in K-free medium, the half-times of K efflux and Na influx in young rat cells were approximately 30 min, significantly higher than the half-time in the presence of serum, and with ouabain, being approximately 13 min. Histograms of distributions of K and Na content showed that the cells behaved as a single functional population. Ouabain Ki was estimated to be 10(-4) M. After 24 h preincubation in K-free medium, when returned in 5 mM K-containing medium, adult rat cells recovered rapidly normal intracellular K and Na concentrations. Using this approach, expression of the kinetics ionic transport properties of renal epithelial cells during development, and the hormonal influences on terminal differentiation may be studied.
从成年和幼年大鼠最外层的浅表皮质中分离出肾上皮细胞。这些细胞可能起源于近端，接种在硅片上，并培养1 - 3天。使用电子探针分析，在小群体的单个细胞中测量细胞内钾、钠、氯和磷的含量及浓度，以及在用哇巴因孵育或在无钾培养基中抑制钠钾ATP酶后细胞内含量变化的动力学。在对照培养基中，以毫摩尔为单位的浓度大致为：钾，130；钠，15；氯，28；磷，140。在抑制钠钾ATP酶6小时后，细胞将所有的钾交换为钠，在无钾培养基中细胞内钠浓度增加到139毫摩尔。氯浓度最多增加到46毫摩尔。在无钾培养基中孵育24小时后，细胞内钾 + 钠 + 氯的总和增加不超过25%。在相似条件下，成年和幼年大鼠细胞的细胞内钾、钠和氯没有差异。在成年大鼠中测量到抑制钠钾ATP酶后钾外流和钠内流的半衰期约为16 - 20分钟。在无血清的无钾培养基中，幼年大鼠细胞钾外流和钠内流的半衰期约为30分钟，显著高于有血清时的半衰期，在用哇巴因处理时约为13分钟。钾和钠含量分布的直方图表明，这些细胞表现为单一功能群体。哇巴因的抑制常数估计为10(-4) M。在无钾培养基中预孵育24小时后，当重新置于含5毫摩尔钾的培养基中时，成年大鼠细胞迅速恢复正常的细胞内钾和钠浓度。使用这种方法，可以研究肾上皮细胞在发育过程中离子转运特性的动力学表达以及激素对终末分化的影响。
