Laboratory of Animal Biotechnology, Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Av. Vicente Mendez 595, Chillan 3780000, Chile.
ANID-Millennium Science Initiative Program Millennium Nucleus of Ion Channels-Associated Diseases (MiNICAD), Center for Bioinformatics, Simulation and Modeling, CBSM, Department of Bioinformatics, Faculty of Engineering, Campus Talca, University of Talca, Talca 3460000, Chile.
Int J Mol Sci. 2023 Apr 18;24(8):7438. doi: 10.3390/ijms24087438.
The embryo-maternal interaction occurs during the early stages of embryo development and is essential for the implantation and full-term development of the embryo. In bovines, the secretion of interferon Tau (IFNT) during elongation is the main signal for pregnancy recognition, but its expression starts around the blastocyst stage. Embryos release extracellular vesicles (EVs) as an alternative mechanism of embryo-maternal communication. The aim of the study was to determine whether EVs secreted by bovine embryos during blastulation (D5-D7) could induce transcriptomic modifications, activating IFNT signaling in endometrial cells. Additionally, it aims to assess whether the EVs secreted by embryos produced in vivo (EVs-IVV) or in vitro (EVs-IVP) have different effects on the transcriptomic profiles of the endometrial cells. In vitro- and in vivo-produced bovine morulae were selected and individually cultured for 48 h to collect embryonic EVs (E-EVs) secreted during blastulation. E-EVs stained with PKH67 were added to in vitro-cultured bovine endometrial cells to assess EV internalization. The effect of EVs on the transcriptomic profile of endometrial cells was determined by RNA sequencing. EVs from both types of embryos induced several classical and non-classical IFNT-stimulated genes (ISGs) and other pathways related to endometrial function in epithelial endometrial cells. Higher numbers of differentially expressed genes (3552) were induced by EVs released by IVP embryos compared to EVs from IVV (1838). Gene ontology analysis showed that EVs-IVP/IVV induced the upregulation of the extracellular exosome pathway, the cellular response to stimulus, and the protein modification processes. This work provides evidence regarding the effect of embryo origin (in vivo or in vitro) on the early embryo-maternal interaction mediated by extracellular vesicles.
胚胎-母体相互作用发生在胚胎发育的早期阶段，对于胚胎的着床和足月发育至关重要。在牛中，伸长过程中干扰素 Tau（IFNT）的分泌是妊娠识别的主要信号，但它的表达始于囊胚阶段。胚胎通过释放细胞外囊泡（EVs）作为胚胎-母体通讯的替代机制。本研究旨在确定在囊胚期（D5-D7）期间，牛胚胎分泌的 EV 是否可以诱导转录组修饰，激活子宫内膜细胞中的 IFNT 信号。此外，还旨在评估体内（EV-IVV）或体外（EV-IVP）产生的胚胎分泌的 EV 是否对子宫内膜细胞的转录组谱有不同的影响。选择体内和体外产生的牛桑葚胚并分别培养 48 小时，以收集囊胚期分泌的胚胎 EV（E-EVs）。用 PKH67 染色的 E-EVs 添加到体外培养的牛子宫内膜细胞中，以评估 EV 的内化。通过 RNA 测序确定 EV 对子宫内膜细胞转录组谱的影响。来自两种胚胎的 EV 诱导了上皮子宫内膜细胞中几个经典和非经典的 IFNT 刺激基因（ISGs）和其他与子宫内膜功能相关的途径。与来自 IVV 的 EV（1838 个）相比，来自 IVP 胚胎的 EV 诱导了更多数量的差异表达基因（3552 个）。基因本体分析表明，EV-IVP/IVV 诱导了细胞外外泌体途径、细胞对刺激的反应和蛋白质修饰过程的上调。这项工作提供了关于胚胎起源（体内或体外）对细胞外囊泡介导的早期胚胎-母体相互作用的影响的证据。
