Chandler C E, Cragoe E J, Glaser L
J Cell Physiol. 1985 Dec;125(3):367-78. doi: 10.1002/jcp.1041250303.
We have reexamined the possible role of the Na+/H+ antiport in the cellular response by PC12 pheochromocytoma cells to nerve growth factor (NGF). In contrast to previous reports, we observe no activation of Na+/H+ exchange in these cells, using a very sensitive assay based on the measurement of cytoplasmic pH with dimethylfluorescein dextran (Rothenberg et al., J. Biol. Chem., 258:4883-4809, 1983). Our measurements indicate that the PC12 pheochromocytoma cells, under all conditions tested, show a high rate of Na+/H+ exchange. The discrepancy between these observations and previous experiments could be due to differences in cells in different laboratories, but also to changes in cell adhesion induced by NGF. We describe conditions where intracellular pH and rates of Na+ uptake can be measured reliably in PC12 cells with adequate controls for cell adhesion. We conclude that activation of Na+/H+ exchange is neither sufficient nor required for the differentiation of PC12 cells induced by NGF.
我们重新审视了Na⁺/H⁺逆向转运体在PC12嗜铬细胞瘤细胞对神经生长因子(NGF)的细胞反应中可能发挥的作用。与之前的报道相反,我们采用基于用二甲基荧光素葡聚糖测量细胞质pH值的非常灵敏的检测方法(Rothenberg等人,《生物化学杂志》,258:4883 - 4809,1983),发现在这些细胞中未观察到Na⁺/H⁺交换的激活。我们的测量结果表明,在所有测试条件下,PC12嗜铬细胞瘤细胞都显示出较高的Na⁺/H⁺交换速率。这些观察结果与之前实验之间的差异可能是由于不同实验室所用细胞的差异,也可能是由于NGF诱导的细胞黏附变化。我们描述了在PC12细胞中能够可靠测量细胞内pH值和Na⁺摄取速率的条件,同时对细胞黏附进行了充分的对照。我们得出结论,对于NGF诱导的PC12细胞分化而言,Na⁺/H⁺交换的激活既不是充分条件也不是必要条件。