Previous studies have shown that running exercise, either alone or in combination with antidepressant treatment, results in increased hippocampal BDNF levels. Nitric oxide (NO) is an important signaling molecule that has neuronal survival-promoting properties and has been shown to play an important role in plasticity associated with activating interventions. Herein, we administered the NO synthase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), in conjunction with the monoamine oxidase inhibitor (MAOI) antidepressant, tranylcypromine, and voluntary wheel-running exercise to determine whether the enhancement in full-length BDNF mRNA occurring with these interventions is dependent upon NO synthesis. Our results demonstrate that both chronic exercise and chronic exercise-plus-tranylcypromine lead to enhanced hippocampal BDNF mRNA and protein expression. NOS inhibition prevents this effect of chronic exercise, but only partly prevents the effects of the exercise/antidepressant combination. Thus, the robust enhancement in BDNF mRNA occurring with exercise appears to be NO synthesis-dependent, but the intervention including antidepressant may enhance BDNF expression through alternative intracellular mechanisms. In addition, because exercise and antidepressants have both been shown to activate survival-promoting genes, we evaluated the levels of hippocampal phosphatidylinositol 3' kinase (PI-3K), an important signaling molecule within a principal neuronal survival-promoting intracellular pathway. Like BDNF mRNA and protein, exercise increases the expression of PI-3K, whereas concomitant NOS inhibition prevents this increase in PI-3K immunoreactivity above control levels. Our results are discussed in light of possible overlapping, but distinct intracellular pathways activated by exercise and antidepressant treatment to bring about enhancements in BDNF expression and other survival-promoting effects. These findings further demonstrate the potential therapeutic potential of chronic exercise to supplement pharmacotherapeutic treatment of mood disorders.