Nerve growth factor does not activate Na+/H+ exchange in PC12 pheochromocytoma cells

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.