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6-Nitrodopamine is a novel catecholamine released by vascular tissues, heart, and vas deferens. The aim of this study was to investigate whether 6-nitrodopamine is released from the thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) and to evaluate the relaxing and anti-contractile actions of this catecholamine. Release of 6-nitrodopamine, dopamine, noradrenaline, and adrenaline was assessed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The relaxations induced by 6-nitrodopamine and by the selective dopamine D2 receptor antagonist L-741,626 were evaluated on U-46619 (3 nM)-pre-contracted vessels. The effects of 6-nitrodopamine and L-741,626 on the contractions induced by electric-field stimulation (EFS), dopamine, noradrenaline, and adrenaline were also investigated. Both aorta and pulmonary artery rings exhibited endothelium-dependent release of 6-nitrodopamine, which was significantly reduced by the NO synthesis inhibitor L-NAME. Addition of 6-nitrodopamine or L-741,626 caused concentration-dependent relaxations of both vascular tissues, which were almost abolished by endothelium removal, whereas L-NAME and the soluble guanylate cyclase inhibitor ODQ had no effect on 6-nitrodopamine-induced relaxations. Additionally, pre-incubation with 6-nitrodopamine antagonized the dopamine-induced contractions, without affecting the noradrenaline- and adrenaline-induced contractions. Pre-incubation with L-741,626 antagonized the contractions induced by all catecholamines. The EFS-induced contractions were significantly increased by L-NAME, but unaffected by ODQ. Immunohistochemical assays showed no immunostaining of the neural tissue markers S-100 and calretinin in either vascular tissue. The results indicated that 6-nitrodopamine is the major catecholamine released by marmoset vascular tissues, and it acts as a potent and selective antagonist of dopamine D2-like receptors. 6-nitrodopamine release may be the major mechanism by which NO causes vasodilatation. 6Nitrodopamine Nitrodopamine 6 tissues heart deferens 6nitrodopamine nitrodopamine Callithrix spp. spp anticontractile anti contractile 6nitrodopamine, nitrodopamine, noradrenaline LCMS/MS. LCMSMS LC MS/MS . MS (LC-MS/MS) D L741,626 L741626 L 741,626 741 626 L-741,62 U46619 U 46619 U-4661 3 ( nMprecontracted nM pre contracted vessels electricfield electric field EFS, EFS , (EFS) investigated endotheliumdependent dependent LNAME. LNAME NAME. NAME concentrationdependent concentration removal 6nitrodopamineinduced nitrodopamineinduced Additionally preincubation incubation dopamineinduced adrenalineinduced Preincubation Pre catecholamines EFSinduced LNAME, NAME, S100 S 100 S-10 D2like Dlike like receptors vasodilatation LCMS LCMS/MS MSMS (LC-MS/MS L741 L741,62 L74162 741626 741,62 74 62 L-741,6 U4661 4661 U-466 (EFS S10 10 S-1 L74 L741,6 L7416 74162 741,6 7 L-741, U466 466 U-46 S1 1 S- L7 L741, 7416 741, L-741 U46 46 U-4 L-74 U4 4 U- L-7 L-