Research into the effects of nerve growth factor (NGF) has involved study of either the signal transduction process or the morphological result of growth factor treatment (cell proliferation and/or differentiation). The Cytosensor Microphysiometer, a silicon-based biosensor system that allows the continuous and real-time monitoring of extracellular acidification rate changes of cells, was used to study the response of PC12 cells to NGF. Stimulation resulted in a rapid increase in the acidification rate of cells in a concentration-dependent fashion (0.1-200 ng/ml NGF; mean effective concentration value of 153 +/- 54 pM). Inhibition of the NGF receptor-linked protein tyrosine kinase by either genistein or K252a attenuated the acidification rate response to NGF. In addition, the acidification response to NGF could be modified by inhibiting Na+/H+ exchange and, separately, glycolysis. This implicates these processes in the metabolic response of PC12 cells to NGF stimulation.