High fructose consumption induces increases in sympathetic nerve activity, although the underlying cellular mechanism is not fully understood. ATP is essential for neuronal activity. We verified in the present study the hypothesis that fructose directly increases ATP production in catecholamine-containing neuron to cause the release of neurotransmitter. Application of different concentrations of fructose (12.5, 25, 50 or 100 μmole/L) to the mouse neuroblastama N2A cells for 3 days resulted in a dose-related increase in ATP production in the cell. In addition, reactive oxygen species (ROS) production was also significantly increased in a dose-dependent manner. Oxygen consumption rate and proton production of mitochondria were significantly increased under high fructose status. Dopamine level in the medium was increased in N2A cells exposed to fructose. The expression of tyrosine hydroxylase and surface expression of dopamine transporter were also significantly increased. Moreover, expression of glutamate transporter 2 (GLUT2), a fructose transport, was significantly increased in response to fructose treatment. Down-regulation of GLUT2 by transfection of Glut2 siRNA effectively suppressed the induced GLUT2 expression and attenuated fructose-promoted ATP and ROS production, and dopamine secretion from the N2A cells. The fructose-induced dopamine secretion was also blunted by scavenging ROS with mitoTEMPO. Together our data indicated that fructose may directly increases neuronal activity through increase in ATP and ROS production via activation of GLUT2.