Our previous study demonstrated that intracranial self-stimulation of the medial forebrain bundle can increase the in vivo synthesis turnover rate of dopamine (DA) and serotonin (5-HT) in the nucleus accumbens of adrenal-intact rats. The present study examined using microdialysis whether such increases in DA and 5-HT syntheses are influenced by adrenal hormones, which are also activated following intracranial self-stimulation. A decarboxylase inhibitor, NSD-1015, was perfused through reversed microdialysis which enabled the simultaneous measurement of 3,4-dihydroxyphenylalanine (DOPA) and 5-hydroxytryptophan (5-HTP) as an index of the in vivo turnover rate of DA and 5-HT syntheses. Adrenalectomy (ADX) attenuated significantly the self-stimulation-induced increase in dialysate levels of DOPA but not 5-HTP. Corticosterone (Cort) replacement reversed the attenuation in DOPA levels in adrenalectomized rats. The finding indicates that activation of DA synthesis in vivo in the nucleus accumbens during intracranial self-stimulation is dependent on, whereas that of 5-HT synthesis is independent of glucocorticoid modulation.