Reproductive Medicine Center, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu, People's Republic of China.
Reproductive Medicine Centre, Nanjing Drum Tower Hospital Clinical College of Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.
Reprod Biol. 2022 Jun;22(2):100646. doi: 10.1016/j.repbio.2022.100646. Epub 2022 May 11.
The birthweight after frozen embryo transfer (FET) was significantly higher compared with fresh embryo transfer (fresh ET), while the mechanism remains unclear. In this study, we transferred vitrified-warmed or fresh mice blastocysts into pseudopregnant recipients (n = 11 each group) produced by natural mating to avoid the influence of superovulation. The fetal weight, placental weight, placental efficiency and placental architecture were studied at E18.5. Placental RNA-Seq analysis was used to identify candidate different lncRNAs and mRNAs between the FET group and the fresh ET group. We found that the fetal weight was increased in the FET group, with increased placental efficiency and the proportion of placental function related labyrinth zone area. 554 lncRNAs and 1012 mRNAs were differentially expressed. KEGG and GO enrichment analyses showed these differentially expressed lncRNAs and their targeted mRNAs might be related to placental morphogenesis. Furthermore, the most differentially expressed 15 lncRNAs and 15 mRNAs were validated by qRT-PCR, we found the LncRNA embryonic stem cells expressed 1 (Lncenc1) was significantly decreased, and Gjb5, which played an important role in labyrinth zone development, was increased. Gjb5 protein increase was further confirmed by Western blot. Lncenc1 and Gjb5 had 48 predicted co-targeted miRNAs, while the correlation analysis of Lncenc1 and Gjb5 mRNA showed a significant inverse correlation. The results showed that FET treatment might enhance the placental function to increase mouse fetal weight via the network diagram of Lncenc1-miRNA-Gjb5.
冻融胚胎移植(FET)后的出生体重明显高于新鲜胚胎移植(新鲜 ET),但其机制尚不清楚。在这项研究中,我们通过自然交配将玻璃化冷冻解冻或新鲜的小鼠囊胚移植到假孕受体中(每组各 11 个),以避免超排卵的影响。在 E18.5 时研究了胎儿体重、胎盘重量、胎盘效率和胎盘结构。利用胎盘 RNA-Seq 分析鉴定 FET 组和新鲜 ET 组之间候选差异 lncRNA 和 mRNA。我们发现 FET 组胎儿体重增加,胎盘效率增加,与胎盘功能相关的绒毛区比例增加。有 554 个 lncRNA 和 1012 个 mRNA 表达差异。KEGG 和 GO 富集分析表明,这些差异表达的 lncRNA 和它们的靶向 mRNA 可能与胎盘形态发生有关。此外,通过 qRT-PCR 验证了最具差异表达的 15 个 lncRNA 和 15 个 mRNA,发现胚胎干细胞表达 1(Lncenc1)显著降低,而在绒毛区发育中起重要作用的 Gjb5 增加。Western blot 进一步证实了 Gjb5 蛋白的增加。Lncenc1 和 Gjb5 有 48 个预测的共同靶向 miRNA,而 Lncenc1 和 Gjb5 mRNA 的相关性分析显示出显著的负相关。结果表明,FET 处理可能通过 Lncenc1-miRNA-Gjb5 网络图增强胎盘功能,从而增加小鼠胎儿体重。