Xiao Longfei, Hu Junjie, Zhao Xingxu, Song Liangli, Zhang Yong, Dong Weitao, Zhang Quanwei, Ma Youji, Li Fadi
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.
College of Life Science and Technology, Gansu Agricultural University, Lanzhou, China.
Reprod Domest Anim. 2018 Oct;53(5):1142-1148. doi: 10.1111/rda.13218. Epub 2018 Jun 25.
Melatonin is an important factor involved in regulating reproduction; it is synthesized enzymatically by the sequential action of melatonin-synthesizing enzymes, arylalkylamine N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT), and exerts its biological functions mainly through receptor-mediated action. To evaluate the expression of melatonin, two melatonin-synthesizing enzymes (HIOMT and AANAT), and membrane receptors (MT1 and MT2) in oestrous corpus luteum (CL) and CL verum of sheep (Ovis aries), we performed ELISA, qRT-PCR, western blotting and immunohistochemistry. The quantitative results showed that melatonin, HIOMT and AANAT levels in the CL verum were significantly higher than those in oestrous CL (p < 0.05), whereas MT1 and MT2 exhibited no change between the oestrous CL and CL verum (p > 0.05); moreover, the localization results showed that HIOMT, AANAT, MT1 and MT2 were mainly expressed in large luteal cells (LLCs). In summary, the above results suggested that sheep CL has potential for the synthesis of melatonin; meanwhile, they also suggested that CL is one of the targets of melatonin. These results provide not only a basis for whether sheep CL can synthesize melatonin but also provide a reference for further study on the mechanism of melatonin in the CL.
褪黑素是参与调节生殖的一个重要因素;它通过褪黑素合成酶、芳基烷基胺N - 乙酰基转移酶(AANAT)和羟基吲哚 - O - 甲基转移酶(HIOMT)的顺序作用酶促合成,并主要通过受体介导的作用发挥其生物学功能。为了评估褪黑素、两种褪黑素合成酶(HIOMT和AANAT)以及膜受体(MT1和MT2)在绵羊(Ovis aries)发情期黄体(CL)和真黄体中的表达,我们进行了酶联免疫吸附测定(ELISA)、实时定量聚合酶链反应(qRT - PCR)、蛋白质印迹法和免疫组织化学。定量结果显示,真黄体中褪黑素、HIOMT和AANAT的水平显著高于发情期黄体(p < 0.05),而MT1和MT2在发情期黄体和真黄体之间没有变化(p > 0.05);此外,定位结果表明,HIOMT、AANAT、MT1和MT2主要在大黄体细胞(LLCs)中表达。总之,上述结果表明绵羊黄体具有合成褪黑素的潜力;同时,它们还表明黄体是褪黑素的作用靶点之一。这些结果不仅为绵羊黄体是否能合成褪黑素提供了依据,也为进一步研究褪黑素在黄体中的作用机制提供了参考。