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      Melatonin MT 1 and MT 2 Receptors in the Ram Reproductive Tract

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          Abstract

          Some melatonin functions in mammals are exerted through MT 1 and MT 2 receptors. However, there are no reports of their presence in the reproductive tract of the ram, a seasonal species. Thus, we have investigated their existence in the ram testis, epididymis, accessory glands and ductus deferens. Real-time polymerase chain reaction (qPCR) revealed higher levels of m-RNA for both receptors in the testis, ampulla, seminal vesicles, and vas deferens, than in the other organs of the reproductive tract ( p < 0.05). Western blot analyses showed protein bands compatible with the MT 1 in the testis and cauda epididymis, and for the MT 2 in the cauda epididymis and deferent duct. Immunohistochemistry analyses revealed the presence of MT 1 receptors in spermatogonias, spermatocytes, and spermatids, and MT 2 receptors in the newly-formed spermatozoa in the testis, whereas both receptors were located in the epithelial cells of the ampulla, seminal vesicles, and ductus deferens. Indirect immunofluorescence showed significant differences in the immunolocation of both receptors in spermatozoa during their transit in the epididymis. In conclusion, it was demonstrated that melatonin receptors are present in the ram reproductive tract. These results open the way for new studies on the molecular mechanism of melatonin and the biological significance of its receptors.

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          Actions of melatonin in the reduction of oxidative stress: A review

          Russel Reiter, Dun-xian Tan, Carmen Osuna … (2000)
          Melatonin was discovered to be a direct free radical scavenger less than 10 years ago. Besides its ability to directly neutralize a number of free radicals and reactive oxygen and nitrogen species, it stimulates several antioxidative enzymes which increase its efficiency as an antioxidant. In terms of direct free radical scavenging, melatonin interacts with the highly toxic hydroxyl radical with a rate constant equivalent to that of other highly efficient hydroxyl radical scavengers. Additionally, melatonin reportedly neutralizes hydrogen peroxide, singlet oxygen, peroxynitrite anion, nitric oxide and hypochlorous acid. The following antioxidative enzymes are also stimulated by melatonin: superoxide dismutase, glutathione peroxidase and glutathione reductase. Melatonin has been widely used as a protective agent against a wide variety of processes and agents that damage tissues via free radical mechanisms.
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            The changing biological roles of melatonin during evolution: from an antioxidant to signals of darkness, sexual selection and fitness.

            Dun-Xian Tan, Rüdiger Hardeland, Lucien Manchester … (2010)
            Melatonin is a molecule present in a multitude of taxa and may be ubiquitous in organisms. It has been found in bacteria, unicellular eukaryotes, macroalgae, fungi, plants and animals. A primary biological function of melatonin in primitive unicellular organisms is in antioxidant defence to protect against toxic free radical damage. During evolution, melatonin has been adopted by multicellular organisms to perform many other biological functions. These functions likely include the chemical expression of darkness in vertebrates, environmental tolerance in fungi and plants, sexual signaling in birds and fish, seasonal reproductive regulation in photoperiodic mammals, and immunomodulation and anti-inflammatory activity in all vertebrates tested. Moreover, its waning production during aging may indicate senescence in terms of a bio-clock in many organisms. Conversely, high melatonin levels can serve as a signal of vitality and health. The multiple biological functions of melatonin can partially be attributed to its unconventional metabolism which is comprised of multi-enzymatic, pseudo-enzymatic and non-enzymatic pathways. As a result, several bioactive metabolites of melatonin are formed during its metabolism and some of the presumed biological functions of melatonin reported to date may, in fact, be mediated by these metabolites. The changing biological roles of melatonin seem to have evolved from its primary function as an antioxidant.
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              The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization.

              Angélique Levoye, Mohammed A. Ayoub, Jean-Luc Guillaume … (2006)
              One-third of the approximately 400 nonodorant G protein-coupled receptors (GPCRs) are still orphans. Although a considerable number of these receptors are likely to transduce cellular signals in response to ligands that remain to be identified, they may also have ligand-independent functions. Several members of the GPCR family have been shown to modulate the function of other receptors through heterodimerization. We show that GPR50, an orphan GPCR, heterodimerizes constitutively and specifically with MT(1) and MT(2) melatonin receptors, using biochemical and biophysical approaches in intact cells. Whereas the association between GPR50 and MT(2) did not modify MT(2) function, GPR50 abolished high-affinity agonist binding and G protein coupling to the MT(1) protomer engaged in the heterodimer. Deletion of the large C-terminal tail of GPR50 suppressed the inhibitory effect of GPR50 on MT(1) without affecting heterodimerization, indicating that this domain regulates the interaction of regulatory proteins to MT(1). Pairing orphan GPCRs to potential heterodimerization partners might be of clinical importance and may become a general strategy to better understand the function of orphan GPCRs.
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                Author and article information

                Contributors
                Russel J. Reiter: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Int J Mol Sci
                Journal ID (iso-abbrev): Int J Mol Sci
                Journal ID (publisher-id): ijms
                Title: International Journal of Molecular Sciences
                Publisher: MDPI
                ISSN (Electronic): 1422-0067
                Publication date (Electronic): 19 March 2017
                Publication date Collection: March 2017
                Volume: 18
                Issue: 3
                Electronic Location Identifier: 662
                Affiliations
                [1 ]Grupo Biología y Fisiología de la Reproducción, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain; martagonzalezarto@ 123456hotmail.com (M.G.-A.); d.aguilar27@ 123456hotmail.com (D.A.); anaelenagaspar@ 123456gmail.com (E.G.-T.); e.serranoblesa@ 123456gmail.com (E.S.-B.); rosperez@ 123456unizar.es (R.P.-P.); muino@ 123456unizar.es (T.M.-B.); pcebrian@ 123456unizar.es (J.C.-P.)
                [2 ]Departamento de Patología Animal, Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain; mgallego@ 123456unizar.es
                [3 ]Departamento de Producción Animal, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, 11001 Bogotá, Colombia; mcarvajals@ 123456unal.edu.co
                [4 ]Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Veterinaria, Instituto de Investigación Sanitaria de Aragón-Universidad de Zaragoza, 50013 Zaragoza, Spain; l.vte.herrera@ 123456gmail.com
                [5 ]Dpto. de Reprodução Animal, da Faculdade de Medicina Veterinaria e Zootecnia, da Universidade de São Paulo, 05508 270 São Paulo, Brazil; thaisroseh@ 123456gmail.com
                Author notes
                [* ]Correspondence: adriana@ 123456unizar.es ; Tel.: +34-976-761-643
                Article
                Publisher ID: ijms-18-00662
                DOI: 10.3390/ijms18030662
                PMC ID: 5372674
                PubMed ID: 28335493
                SO-VID: 0bb16f60-21fc-4678-9be4-44c1cd3bbcdd
                Copyright © © 2017 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 21 February 2017
                Date accepted : 15 March 2017
                Categories
                Subject: Article

                ScienceOpen disciplines: Molecular biology
                Keywords: melatonin receptor,ram,testis,epididymis,accessory glands
                Data availability:
                ScienceOpen disciplines: Molecular biology
                Keywords: melatonin receptor, ram, testis, epididymis, accessory glands

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