The pivotal role of neuronal nitric oxide synthase in the release of 6-nitrodopamine from mouse isolated vas deferens

José Britto-Júnior; Samuel Goulart Nacário Silva; Antonio Tiago Lima; Vivian Fuguhara; Larissa Bueno Andrade; Gustavo Duarte Mendes; Larryn W Peterson; Silvana Chiavegatto; Edson Antunes; Gilberto De Nucci

doi:10.1016/j.niox.2023.12.002

The pivotal role of neuronal nitric oxide synthase in the release of 6-nitrodopamine from mouse isolated vas deferens

Nitric Oxide. 2024 Feb 1:143:1-8. doi: 10.1016/j.niox.2023.12.002. Epub 2023 Dec 12.

Affiliations

¹ Faculty of Medical Sciences, Department of Pharmacology, University of Campinas (UNICAMP), Campinas, Brazil. Electronic address: josebrittojr@dac.unicamp.br.
² Faculty of Medical Sciences, Department of Pharmacology, University of Campinas (UNICAMP), Campinas, Brazil.
³ Department of Pharmacology, Faculty of Medicine, São Leopoldo Mandic, Campinas, SP, Brazil; Department of Pharmacology, Faculty of Medicine, Metropolitan University of Santos, Santos, SP, Brazil.
⁴ Department of Chemistry, Rhodes College, Memphis, TN, USA.
⁵ Department of Pharmacology, Institute of Biomedical Sciences (ICB), University of Sao Paulo (USP), São Paulo, Brazil; Department of Psychiatry, Institute of Psychiatry (IPq), University of Sao Paulo Medical School (FMUSP), São Paulo, Brazil.
⁶ Faculty of Medical Sciences, Department of Pharmacology, University of Campinas (UNICAMP), Campinas, Brazil; Department of Pharmacology, Faculty of Medicine, São Leopoldo Mandic, Campinas, SP, Brazil; Department of Pharmacology, Faculty of Medicine, Metropolitan University of Santos, Santos, SP, Brazil; Department of Pharmacology, Institute of Biomedical Sciences (ICB), University of Sao Paulo (USP), São Paulo, Brazil.

PMID: 38096948
DOI: 10.1016/j.niox.2023.12.002

Abstract

6-Nitrodopamine (6-ND) is released from rat and human vas deferens and is considered a major mediator of both tissues contractility. The contractions induced by 6-ND are selectively blocked by both tricyclic antidepressants and α₁-adrenoceptor antagonists. Endothelial nitric oxide synthase (eNOS) is the major isoform responsible for 6-ND release in mouse isolated heart, however the origin of 6-ND in the vas deferens is unknown. Here it was investigated by LC-MS/MS the basal release of 6-ND from isolated vas deferens obtained from control, eNOS^-/-, nNOS^-/-, and iNOS^-/- mice. In addition, it was evaluated in vitro vas deferens contractility following electric field stimulation (EFS). Basal release of 6-ND was significantly reduced in nNOS^-/- mice compared to control mice, but not decreased when the vas deferens were obtained from either eNOS^-/- or iNOS^-/- mice. Pre-incubation of the vas deferens with tetrodotoxin (1 μM) significantly reduced the basal release of 6-ND from control, eNOS^-/-, and iNOS^-/- mice but had no effect on the basal release of 6-ND from nNOS^-/- mice. EFS-induced frequency-dependent contractions of the vas deferens, which were significantly reduced when the tissues obtained from control, eNOS^-/- and iNOS^-/- mice, were pre-incubated with l-NAME, but unaltered when the vas deferens was obtained from nNOS^-/- mice. In addition, the EFS-induced contractions were significantly smaller when the vas deferens were obtained from nNOS^-/- mice. The results clearly demonstrate that nNOS is the main NO isoform responsible for 6-ND release in mouse vas deferens and reinforces the concept of 6-ND as a major modulator of vas deferens contractility.

Keywords: Adrenaline; Catecholamine; Dopamine; Liquid chromatography; Noradrenaline; Tandem mass spectrometry.

MeSH terms

Animals
Chromatography, Liquid
Dopamine* / analogs & derivatives
Humans
Male
Mice
Muscle Contraction
Nitric Oxide / pharmacology
Nitric Oxide Synthase Type I
Norepinephrine* / pharmacology
Rats
Tandem Mass Spectrometry
Vas Deferens* / physiology

Substances

6-nitrodopamine
Dopamine
Nitric Oxide
Nitric Oxide Synthase Type I
Norepinephrine
Nos1 protein, mouse