Department of Obstetrics and Gynecology, Juntendo University Faculty of Medicine, 2-1-1 Hongo, Bunkyoku, Tokyo 113-8421, Japan.
Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan.
Placenta. 2018 May;65:47-54. doi: 10.1016/j.placenta.2018.04.001. Epub 2018 Apr 7.
In mammals, the placenta is an organ that is required to maintain the development of fetus during pregnancy. Although the proper formation of placenta is in part regulated by the post-translational modifications of proteins, little is known regarding protein arginine methylation during placental development. Here, we characterized developmental expression of protein arginine methyltransferase 1 (PRMT1) in mouse placentas.
Expression levels of PRMT1 mRNA and protein in placentas were investigated using the real-time quantitative PCR and Western blot, respectively. Next, the localization of PRMT1 was determined by immunohistochemistry and immunofluorescence analyses. In addition, the levels of methylarginines of placental proteins were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
PRMT1 mRNA and its protein were expressed at highest levels in mid-gestation stages, and their expression showed stepwise decrease in the late gestation. At embryonic (E) day 9, PRMT1 was observed in several different trophoblast cell (TC) subtypes. Furthermore, PRMT1 was mainly expressed in the labyrinth zone of TCs at E13. Finally, total methylarginines of proteins were significantly reduced in late gestation of placentas compared with mid-gestation stages.
In this study, we found developmental changes in the placental expression of PRMT1 and in protein arginine methylation status during pregnancy. These findings provide fundamental information regarding placental PRMT1-mediated arginine methylation during the development.
在哺乳动物中,胎盘是一种在怀孕期间维持胎儿发育所必需的器官。尽管蛋白质的翻译后修饰在一定程度上调节了胎盘的适当形成,但对于胎盘发育过程中的蛋白质精氨酸甲基化知之甚少。在这里,我们描述了蛋白精氨酸甲基转移酶 1(PRMT1)在小鼠胎盘中的发育表达。
使用实时定量 PCR 和 Western blot 分别研究 PRMT1 mRNA 和蛋白质在胎盘中的表达水平。接下来,通过免疫组织化学和免疫荧光分析确定 PRMT1 的定位。此外,使用液相色谱-串联质谱(LC-MS/MS)定量胎盘蛋白的甲基精氨酸水平。
PRMT1 mRNA 和其蛋白在妊娠中期表达水平最高,在妊娠晚期呈逐步下降。在胚胎(E)第 9 天,PRMT1 观察到在几种不同的滋养层细胞(TC)亚型中。此外,PRMT1 主要在 E13 的绒毛膜区表达。最后,与妊娠中期相比,胎盘在妊娠晚期的总甲基精氨酸显著减少。
在这项研究中,我们发现 PRMT1 在胎盘中的表达和怀孕期间蛋白质精氨酸甲基化状态发生了发育变化。这些发现为 PRMT1 介导的发育过程中胎盘精氨酸甲基化提供了基本信息。