College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, PR China; National Beef Cattle Improvement Centre of China, Yangling, Shaanxi, PR China.
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, PR China.
Anim Reprod Sci. 2014 Jun 10;147(1-2):10-6. doi: 10.1016/j.anireprosci.2014.03.017. Epub 2014 Mar 31.
Melatonin and its receptors are found in the testis of many species, where they mediate testicular functions. The present study aimed to investigate the expression of melatonin receptors (MT1 and MT2) in bovine Sertoli cells (SCs), using reverse transcription polymerase chain reaction (RT-PCR) and western blot. In addition, we assessed the mRNA levels of spermatogenesis-related genes (real-time PCR) and secretion of inhibin B after treatment with various concentrations (0, 80, 160, and 320 pg/mL) of melatonin at different time points (24, 48, or 72 h). We found that bovine SCs express MT1 and MT2 receptors, which were regulated by melatonin in time- and dose-dependent manners after treatment with melatonin. Exogenous melatonin up-regulated the expression of spermatogenesis-related genes, including Cyclin D1, Cyclin E, Pdgfa, Dhh, Occludin, and Claudin, and decreased the mRNA levels of P21 and Kit1 in a time or dose-dependent manner. Meanwhile, melatonin supplementation significantly affected Inhba, Inhbb and Inha mRNA expression. These findings were consistent with inhibin B levels detected in the culture medium. In conclusion, exogenous melatonin acts via its receptors and appears to play regulatory roles in the development and function of bovine SCs.
褪黑素及其受体存在于许多物种的睾丸中,在那里介导睾丸功能。本研究旨在通过逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法研究褪黑素受体(MT1 和 MT2)在牛睾丸支持细胞(SCs)中的表达。此外,我们评估了不同浓度(0、80、160 和 320 pg/mL)褪黑素在不同时间点(24、48 或 72 h)处理后,与精子发生相关的基因(实时 PCR)的 mRNA 水平和抑制素 B 的分泌。我们发现牛 SC 表达 MT1 和 MT2 受体,褪黑素处理后,这些受体受褪黑素的时间和剂量依赖性调节。外源性褪黑素上调与精子发生相关的基因表达,包括 Cyclin D1、Cyclin E、Pdgfa、Dhh、Occludin 和 Claudin,同时以时间或剂量依赖性方式降低 P21 和 Kit1 的 mRNA 水平。同时,褪黑素补充显著影响 Inhba、Inhbb 和 Inha mRNA 的表达。这些发现与培养基中检测到的抑制素 B 水平一致。总之,外源性褪黑素通过其受体发挥作用,似乎在牛 SC 的发育和功能中发挥调节作用。