González-Arto Marta, Aguilar David, Gaspar-Torrubia Elena, Gallego Margarita, Carvajal-Serna Melissa, Herrera-Marcos Luis V, Serrano-Blesa Edith, Hamilton Thais Rose Dos Santos, Pérez-Pé Rosaura, Muiño-Blanco Teresa, Cebrián-Pérez José A, Casao Adriana
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.
Departamento de Patología Animal, Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
Int J Mol Sci. 2017 Mar 19;18(3):662. doi: 10.3390/ijms18030662.
Some melatonin functions in mammals are exerted through MT₁ and MT₂ 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 ( < 0.05). Western blot analyses showed protein bands compatible with the MT₁ in the testis and cauda epididymis, and for the MT₂ in the cauda epididymis and deferent duct. Immunohistochemistry analyses revealed the presence of MT₁ receptors in spermatogonias, spermatocytes, and spermatids, and MT₂ 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.
褪黑素在哺乳动物中的一些功能是通过MT₁和MT₂受体发挥的。然而,在季节性繁殖的公羊的生殖道中,尚无关于这些受体存在的报道。因此,我们研究了它们在公羊睾丸、附睾、附属腺体和输精管中的存在情况。实时聚合酶链反应(qPCR)显示,睾丸、壶腹、精囊和输精管中这两种受体的mRNA水平高于生殖道的其他器官(P<0.05)。蛋白质印迹分析显示,睾丸和附睾尾部有与MT₁相符的蛋白条带,附睾尾部和输精管中有与MT₂相符的蛋白条带。免疫组织化学分析显示,睾丸中的精原细胞、精母细胞和精子细胞中有MT₁受体,新形成的精子中有MT₂受体,而壶腹、精囊和输精管的上皮细胞中均有这两种受体。间接免疫荧光显示,精子在附睾中转运时,这两种受体的免疫定位存在显著差异。总之,已证明褪黑素受体存在于公羊生殖道中。这些结果为褪黑素分子机制及其受体的生物学意义的新研究开辟了道路。