González-Arto Marta, Vicente-Carrillo Alejandro, Martínez-Pastor Felipe, Fernández-Alegre Estela, Roca Jordi, Miró Jordi, Rigau Teresa, Rodríguez-Gil Joan E, Pérez-Pé Rosaura, Muiño-Blanco Teresa, Cebrián-Pérez José A, Casao Adriana
Grupo Biología y Fisiología de la Reproducción, Facultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Universidad de Zaragoza, Zaragoza, Spain.
Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.
Theriogenology. 2016 Nov;86(8):1958-68. doi: 10.1016/j.theriogenology.2016.06.016. Epub 2016 Jun 23.
Melatonin is a ubiquitous and multipurpose molecule, and one of its roles is to regulate reproduction in some seasonal mammals. Our group has previously reported the variation in the melatonin levels in ram seminal plasma along the year and identified MT1 and MT2 receptors in ram spermatozoa. The objective of this study was to elucidate whether the presence of melatonin receptors (MT1 and MT2) in the sperm plasma membrane, and melatonin in the seminal plasma is related to seasonal breeding. For this purpose, the presence of melatonin receptors and the levels of melatonin in seminal plasma have been examined in several species: donkey and stallion as long-day breeders; red deer as a wild, short-day, highly seasonal breeder (epididymal spermatozoa); bull as a conventional nonseasonal breeder; boar as a seasonal breeder under management techniques; and dog as possible a seasonal breeder not regulated by melatonin. We have detected measurable levels of melatonin in the seminal plasma of all ejaculated semen samples (from donkey, stallion, boar, bull, and dog). Also, and for the first time, we have demonstrated the presence of MT1 and MT2 melatonin receptors in the spermatozoa of all these species, regardless their type of reproduction or sperm source (ejaculated or epididymal), using indirect immunofluorescence techniques and Western blotting. Our findings suggest that melatonin and melatonin receptors may be universally distributed in the reproductive system of mammals and that the sperm melatonin receptors cells may not be necessarily related with seasonal reproduction. Furthermore, the presence of MT1 at the cytoplasmic droplet in immature ejaculated stallion spermatozoa found in one sample and epididymal red deer spermatozoa suggests that melatonin may be involved in specific functions during spermatogenesis and sperm maturation, like protecting spermatozoa from oxidative damage, this activity being mediated through these receptors.
褪黑素是一种广泛存在且具有多种功能的分子,其作用之一是调节一些季节性繁殖哺乳动物的生殖活动。我们团队之前报道了公羊精液中褪黑素水平随季节的变化,并在公羊精子中鉴定出了MT1和MT2受体。本研究的目的是阐明精子质膜中褪黑素受体(MT1和MT2)以及精液中褪黑素的存在是否与季节性繁殖有关。为此,我们检测了多个物种精液中褪黑素受体的存在情况以及褪黑素水平:作为长日照繁殖动物的驴和种马;作为野生、短日照、高度季节性繁殖动物的马鹿(附睾精子);作为传统非季节性繁殖动物的公牛;作为在管理技术下季节性繁殖动物的野猪;以及可能不受褪黑素调节的季节性繁殖动物狗。我们在所有射出精液样本(来自驴、种马、野猪、公牛和狗)的精液中都检测到了可测量水平的褪黑素。此外,我们首次使用间接免疫荧光技术和蛋白质印迹法证明了所有这些物种的精子中都存在MT1和MT2褪黑素受体,无论它们的繁殖类型或精子来源(射出的或附睾的)。我们的研究结果表明,褪黑素和褪黑素受体可能普遍分布于哺乳动物的生殖系统中,精子褪黑素受体细胞不一定与季节性繁殖相关。此外,在一个样本中发现的未成熟射出种马精子的细胞质滴以及附睾马鹿精子中存在MT1,这表明褪黑素可能参与精子发生和精子成熟过程中的特定功能,如保护精子免受氧化损伤,这种活性是通过这些受体介导的。