Abstract
The extracellular vesicles (EVs) of uterine flushing fluids (UFs) mediate intrauterine communication between conceptus and uterus in pigs. The small RNAs of UFs-EVs are widely recognized as important factors that influence embryonic implantation. However, small RNAs expression profiles of porcine UFs-EVs during peri-implantation are still unknown. In this study, cup-shaped EVs of porcine UFs on days 10 (D10), 13 (D13) and 18 (D18) of pregnancy were isolated and characterized. The expression of small RNAs in these EVs was comprehensively profiled through sequencing. A total of 152 known microRNAs (miRNAs), 43 novel miRNAs, 6248 known Piwi-interacting RNAs (piRNAs) and 110 novel piRNAs were identified. Among these small RNAs, RT-qRCR results indicated that ssc-let-7f-5p, ssc-let-7i-5p and ssc-let-7g were differentially expressed during the three stages. Bioinformatics analysis showed that the miRNAs differentially expressed in the three comparisons (D10 vs D13, D13 vs D18 and D10 vs D18) were involved in important processes and pathways related to immunization, endometrial receptivity and embryo development, which play important roles in embryonic implantation. Our results reveal that EVs from porcine UFs contain various small RNAs with potentially vital effects on implantation. This research also provides resources for studies of miRNAs and piRNAs in the cross-talk between embryo and endometrium.
Highlights
The extracellular vesicles (EVs) of uterine flushing fluids (UFs) mediate intrauterine communication between conceptus and uterus in pigs
A previous study has suggested that G-protein coupled receptor may play a vital role in embryo-maternal crosstalk during the period of maternal recognition and establishment of pregnancy [43]. Both previous research and our results reveal that these most highly abundant miRNAs in porcine UFs-EVs may act on regulating embryo development and endometrial receptivity at the time of implantation
The Piwi-interacting RNAs (piRNAs) have been found in spent culture media of embryo [59], our results shown that lots of piRNAs were present in porcine UFs-EVs
Summary
The extracellular vesicles (EVs) of uterine flushing fluids (UFs) mediate intrauterine communication between conceptus and uterus in pigs. Previous studies have indicated that the majority of pre-natal losses occur during the peri-implantation period in pigs [1, 2]. During this period, the porcine blastocyst expands from 0.5–1 mm in diameter at hatching to 2-mm spheres on day of pregnancy (D10) and begins to rapidly elongate into tubular and filamentous shapes on day or [3, 4]. The extracellular vesicles (EVs) of uterine flushing fluids (UFs) are capable of mediating embryo-endometrium cross-talk during implantation [8, 9]. In order to improve the rate of successful implantation in pigs, it is necessary to investigate the mechanism by which intrauterine EVs regulate the cross-talk
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