6-Nitrodopamine Release From Mouse Seminal Vesicles Is Dependent on Endothelial Nitric Oxide Synthase (eNOS) Activation

José Britto-Júnior; Pérola Rafaella Cedano Godoy; Denis Oliveira Lima; Valéria B de Souza; André A Schenka; Fernanda V Mariano; Felipe Fernandes Jacintho; Idam Hermawan; Hiroaki Shimokawa; Masato Tsutsui; Silvana Chiavegatto; Edson Antunes; Gilberto De Nucci

doi:10.1002/prp2.70167

6-Nitrodopamine Release From Mouse Seminal Vesicles Is Dependent on Endothelial Nitric Oxide Synthase (eNOS) Activation

Pharmacol Res Perspect. 2025 Oct;13(5):e70167. doi: 10.1002/prp2.70167.

Authors

Affiliations

¹ Department of Pharmacology, Faculty of Medical Sciences, University of Campinas (UNICAMP), Campinas, Brazil.
² Department of Pathology, Faculty of Medical Science, State University of Campinas (UNICAMP), Campinas, Brazil.
³ Department of Pharmacology, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan.
⁴ Graduate School, International University of Health and Welfare, Narita, Japan.
⁵ Department of Pharmacology, Institute of Biomedical Sciences (ICB), University of Sao Paulo (USP), São Paulo, Brazil.
⁶ Department of Psychiatry, Institute of Psychiatry (IPq), University of Sao Paulo Medical School (FMUSP), São Paulo, Brazil.

Abstract

Human seminal vesicles present basal release of epithelium-derived 6-nitrodopamine (6-ND) and this catecholamine potentiates noradrenaline-induced contractions. Since nitric oxide synthase (NOS) activation is a determining factor involved in 6-ND biosynthesis, this study aimed to investigate which NOS isoform is responsible for the 6-ND release in mouse isolated seminal vesicles (MISV). Male control, eNOS^-/-, NOS^-/-, iNOS^-/-, and e/n/iNOS^-/- mice were used. 6-ND release was evaluated by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). MISV contractility was assessed following electric-field stimulation (EFS) or construction of concentration-response curves to catecholamines. MISV from control, eNOS^-/-, nNOS^-/-, iNOS^-/-, and e/n/iNOS^-/- mice exhibited basal release of 6-ND, but 6-ND release from eNOS^-/- and e/n/iNOS^-/- was significantly reduced compared to control and nNOS^-/- mice. Epithelium removal in MISV from control mice significantly reduced 6-ND levels. EFS (2-32 Hz) induced frequency-dependent MISV contractions in all groups, but the contractions from eNOS^-/- and e/n/iNOS^-/- mice were significantly decreased compared to control groups. In vitro l-NAME treatment or epithelium removal significantly reduced EFS-induced contractions. Pre-incubation of MISV with 6-ND (1-100 nM) significantly potentiated both EFS- and noradrenaline-induced contractions, whereas pre-incubation with noradrenaline, adrenaline, and dopamine did not affect the EFS-induced responses. Immunohistochemistry and fluorescence in situ hybridization (FISH) assays revealed positivity for tyrosine hydroxylase and eNOS in the epithelium and endothelium of the seminal vesicles. In conclusion, MISV releases epithelium-derived 6-ND, and its biosynthesis involves eNOS activation. The finding that 6-ND markedly amplified the noradrenaline-induced contractions indicates that epithelium-derived 6-ND acts as a major modulator of MISV contractility.

Keywords: ejaculation; epithelial cells; nitrocatecholamines; synergism.

MeSH terms

Animals
Dopamine* / analogs & derivatives
Dopamine* / metabolism
Electric Stimulation
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Nitric Oxide Synthase Type I / genetics
Nitric Oxide Synthase Type I / metabolism
Nitric Oxide Synthase Type II / genetics
Nitric Oxide Synthase Type II / metabolism
Nitric Oxide Synthase Type III* / genetics
Nitric Oxide Synthase Type III* / metabolism
Seminal Vesicles* / drug effects
Seminal Vesicles* / metabolism

Substances

Nitric Oxide Synthase Type III
Nos3 protein, mouse
Nitric Oxide Synthase Type I
Nitric Oxide Synthase Type II
Dopamine

Abstract

MeSH terms

Substances

Grants and funding