6-Nitrodopamine is an endogenous mediator of rat seminal vesicles contractility.

Abstract

Background: 6-Nitrodopamine (6-ND) released from rat vas deferens acts an endogenous modulator of vas deferens contractility.

Objectives: To investigate whether rat isolated seminal vesicles (RISV) releases 6-ND, the mechanisms involved in the release, and the modulatory role of 6-ND on tissue contractility.

Methods: Rat seminal vesicles were removed and placed in Krebs-Henseleit's solution at 37°C for 30 min, and an aliquot was used to analyze the concentrations of 6-ND, dopamine, noradrenaline, and adrenaline by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The effect of mechanical removal of the epithelium and the effects of pre-incubation of RISV strips with N^ω-nitro-L-arginine methyl ester (L-NAME; in combination or not with L-arginine), tetrodotoxin (TTX), GKT137831, and hydrogen peroxide (H₂O₂) on 6-ND release were evaluated. For functional studies, RISV strips were mounted in an organ bath and tied to an isometric force transducer. The expressions of tyrosine hydroxylase and endothelial nitric oxide synthase (eNOS, type III NOS) were investigated by immunohistochemistry and/or fluorescence in situ hybridization (FISH).

Results: 6-ND was the major released catecholamine from RISV strips compared with noradrenaline, adrenaline, and dopamine. Epithelium removal significantly reduced the release of 6-ND, noradrenaline and dopamine. In RISV strips obtained from animals chronically treated with L-NAME, the 6-ND release significantly reduced. Pre-incubation with L-NAME reduced 6-ND release, which was partly restored by co-incubation with L-arginine. Pre-incubation with TTX had no effect on the release of any catecholamine, whereas GKT137831 and H₂O₂ significantly increased 6-ND release. All catecholamines produced concentration-dependent RISV contractions, but 6-ND was approximately 30× less potent than the others. 6-ND (0.1 nM) significantly potentiated noradrenaline-, adrenaline-, and dopamine-induced contractions, with such potentiations inhibited by TTX. Immunohistochemistry and FISH assays in RISV tissues identified tyrosine hydroxylase in epithelial cells and eNOS expression in both epithelial and endothelial cells.

Discussion and conclusion: This is the first demonstration that epithelial-derived 6-ND modulates rat seminal vesicle contractility.