Constitutive brain nitric oxide (NO) synthase is described as converting L-arginine into NO. NO is thought to be the cellular messenger released by endothelial cells, and was originally termed endothelial-derived relaxing factor (EDRF). The mechanisms of its synthesis remain unclear. Using microelectrode differential pulse voltammetry in the presence of cortical constitutive rat brain NO synthase, a peak was recorded at -1.66 V with respect to a Ag/AgCl reference electrode. This voltage peak, due to the reduction of N-nitroso-L-arginine, was increased in rat brain cortex after pharmacological stimulation with L-arginine or A-23187; whereas it was abolished following application Of D-arginine, N-nitro-L-arginine or pure NO. Using laser doppler measurements, the secretion of N-nitroso-L-arginine was correlated to brain blood flow. These preliminary results suggest that N-nitroso-L-arginine is synthetized by constitutive brain NO synthase of vascular endothelial cells in rat brain cortex.