鸡胚外泌体miRNAs功能分析

doi:10.11843/j.issn.0366-6964.2020.010

Abstract

Abstract: This study was conducted to elucidate the physiological basis of active substances and their mechanism of chicken embryos based on exosomes and their miRNAs. Eighteen Huiyang Beard chicken embryos with 13 embryo age were selected (3 groups, 6 eggs per group), and embryos-derived exosomes were isolated by enzymatic digestion, sucrose cushion gradient and differential ultracentrifugation, and their characteristics were identified by transmission electron microscopy, nanoflow and labeled protein flow detection. High-throughput sequencing was used to study the expression profile of miRNAs in exosome samples, and target genes prediction and functional analysis were performed for top 10 and co-expressed miRNAs. The results showed that extracellular vesicles with typical characteristics of exosomes were obtained from 3 buoyant density ranges (S1:1.13 g·mL^-1, S2:1.21 g·mL^-1 and S3:1.32 g·mL^-1), respectively. A total of 675, 661 and 845 known chicken miRNAs of S1, S2 and S3 samples were detected via alignment to miRBase V21, in which 488 (52.6%) miRNAs were co-expressed in the 3 samples. These co-expressed miRNAs might participate in the cell proliferation, differentiation, and immune response, etc. Based on both TargetScan 7.2 and miRDB systems, a total of 3 650 genes could be targeted the 56 miRNAs which were top 10 and co-expressed in the 3 samples. In addition, the 3 650 target genes were analyzed by DAVID, it was found that 24 pathways related to growth, development and immunity were significantly enriched by these genes (P<0.05), which including MAPK signaling pathway, mTOR signaling pathway, Notch signaling pathway, Wnt signaling pathway and GnRH signaling pathway, etc. In this study, an effective method to isolate exosomes from chicken embryo tissue was established, and chicken embryos-derived exosome miRNAs could affect biological pathways related to growth, development and immune regulation.

Key words: chicken embryos, exosome, isolation, functional study

CLC Number:

S831.3

LI Ying, WANG Yan, WANG Jie, HE Jingyi, LUO Chenglong, QU Hao. Functional Analysis of microRNAs in Chicken Embryos-derived Exosomes[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1260-1270.

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Functional Analysis of microRNAs in Chicken Embryos-derived Exosomes

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