幼龄和成年太行山羊附睾头差异竞争性内源RNAs机制分析

doi:10.11843/j.issn.0366-6964.2021.05.015

Abstract

Abstract: The aim of this study was to explore and screen the differentially expressed long noncoding RNAs (lncRNAs), microRNAs (miRNAs) and mRNAs in caput epididymis between Taihang goat kids and adult Taihang goats, and to construct a competitive endogenous RNAs (ceRNAs) network associated with the regulatory mechanism of immune-related genes in caput epididymis of Taihang goats. Three 2-month-old healthy Taihang goat kids with similar body weight and three 2-year-old healthy adult Taihang goats with similar body weight were selected in this study. After castration, the caput epididymis were collected for whole transcriptome sequencing. The differentially expressed mRNAs, lncRNAs and miRNAs in caput epididymis were screened between goat kids and adult goats using DESeq2. Based on the principle of ceRNA-score, the differentially expressed ceRNAs was obtained by using miRanda software and R-package (reshape2, dplyr, tidyr). GO and KEGG enrichment analysis were used to study the function of mRNAs in the ceRNAs mechanism. The ceRNAs network related with immune function was drawn in caput epididymis of Taihang goats. Finally, 8 mRNAs, 8 miRNAs and 8 lncRNAs were randomly selected to verify the accuracy of the whole transcriptome sequencing results by qRT-PCR. According to the analysis results, compared with caput epididymis of goat kids, there were 6 461 differentially expressed mRNAs in caput epididymis of adult Taihang goats， including 2 997 up-regulated mRNAs and 3 464 down-regulated mRNAs; There were 1 147 differentially expressed lncRNAs, including 703 up-regulated lncRNAs and 444 down-regulated lncRNAs; There were 182 differentially expressed miRNAs, including 81 up-regulated miRNAs and 101 down-re-gulated miRNAs. Three hundred and sixty-six lncRNAs with ceRNAs regulatory relationship were obtained, including 213 up-regulated lncRNAs and 153 down-regulated lncRNAs; There were 3 131 mRNAs with ceRNAs regulatory relationship, including 1 253 up-regulated mRNAs and 1 878 down-regulated mRNAs; There were 140 miRNAs with ceRNAs regulatory relationship, including 48 up-regulated miRNAs and 92 down-regulated miRNAs. Among the genes in the ceRNAs mechanism, lymphocyte antigen 6 complex locus protein G5B (LY6G5B)，epididymal-specific lipocalin-9 (LCN9), adisintegrinand metalloproteinase 28 (ADAM28) and Mucin 15 (MUC15) were the significantly up-regulated genes related with immune function, which expression in adult goats were higher than those in goat kids (P<0.01). GO and KEGG analysis of the DEGs in the ceRNAs mechanism showed that of differentially expressed genes were involved in protein processing in endoplasmic reticulum, protein export, mucin type O-Glycan biosynthesis, ECM-receptor interaction and other pathways related to cell protein synthesis, secretion and transport. The results of qRT-PCR showed that the expression trends of the differentially expressed mRNAs, lncRNAs and miRNAs selected randomly were consistent with the whole transcriptome sequencing results, except for chi-miR-320-3p. In this study, the analysis results of the immune-related ceRNAs network in caput epididymis showed that lncRNA-MSTRG.22929.11, lncRNA-MSTRG.57822.5, lncRNA-MSTRG.26758.1, lncRNA-MSTRG.12113.3, lncRNA-MSTRG.59930.2 and other lncRNAs were related to immunity and sperm maturation in caput epididymis. In this study, the differentially expressed ceRNAs were screened in caput epididymais between Taihang goat kids and adult Taihang goats. The ceRNAs network related to immunity will provides a foundation for the study of lncRNAs as ceRNAs in the regulation mechanism of epididymal immune function in Taihang goats.

Key words: ceRNA, Taihang goat, caput epididymis, immune

CLC Number:

GUO Xiangqian, DONG Fucheng, HUANG Xinyun, LIU Wenzhong, QIAO Liying, ZHANG Chunxiang, REN Youshe. Analysis of the Differentially Expressed ceRNAs Mechanism in Caput Epididymis between Goat Kids and Adult Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(5): 1293-1306.

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Analysis of the Differentially Expressed ceRNAs Mechanism in Caput Epididymis between Goat Kids and Adult Goats

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