Using exosomal miRNAs extracted from porcine follicular fluid to investigate their role in oocyte development
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RESEARCH ARTICLE
Open Access
Using exosomal miRNAs extracted from porcine follicular fluid to investigate their role in oocyte development Junhe Hu* , Jinyi Dong, Zhi Zeng, Juan Wu, Xiansheng Tan, Tao Tang, Jiao Yan and Chenzhong Jin
Abstracts Background: Follicular development is crucial to normal oocyte maturation, with follicular size closely related to oocyte maturation. To better understand the molecular mechanisms behind porcine oocyte maturation, we obtained exosomal miRNA from porcine follicular fluid (PFF). These miRNA samples were then sequenced and analyzed regarding their different follicular sizes, as described in the methods section. Results: First, these results showed that this process successfully isolated PFF exosomes. Nearly all valid reads from the PFF exosomal sequencing data were successfully mapped to the porcine genome database. Second, we used hierarchical clustering methods to determine that significantly expressed miRNAs were clustered into A, B, C, and D groups in our heatmap according to different follicle sizes. These results allowed for the targeting of potential mRNAs genes related to porcine oocyte development. Third, we chose ten, significantly expressed miRNAs and predicted their target genes for further GO analysis. These results showed that the expression levels of neurotransmitter secretion genes were greatly changed, as were many target genes involved in the regulation of FSH secretion. Notably, these are genes that are very closely related to oocyte maturation in growing follicles. We then used pathway analysis for these targeted genes based on the originally selected ten miRNAs. Results indicated that the pathways were mainly related to the biosynthesis of TGF-beta and its signaling pathway, which are very closely related to reproductive system functions. Conclusions: Finally, these exosomal miRNAs obtained from PFF may provide a valuable addition to our understanding of the mechanism of porcine oocyte maturation. It is also likely that these exosomal miRNAs could function as molecular biomarkers to choose high-quality oocytes and allow for in vitro porcine embryo production. Keywords: Exosomes, Porcine follicular fluid (PFF), Oocyte maturation, Follicular development, miRNA
Background It is well known that oocyte maturation is strongly related to follicular growth and development [1]. As the follicles grow, their size is closely accompanied by increased oocyte diameter, indicating the ever-progressing maturation status of the oocyte. Many past studies have shown that oocyte quality is highly correlated with * Correspondence: [email protected]; [email protected] College of Agriculture and Biotechnology, Hunan University of Humanities, Science and Technology, Road Dingxing 7#, Loudi City 417000, HuNan Province, China
oocyte diameter and later embryo development [1–4]. Moreover, some bovine oocyte maturation increases the volume of follicular fluid, owing to the larger follicle size. This may improve overall oocyte nuclear and cytoplasmic maturation when compared to follicl
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