Abstract

Simple SummaryIn this study, the expression of circRNAs, miRNAs, and mRNA in the immune organs spleen of turbot (Scophthalmus maximus) infected with Aeromonas salmonicida was analyzed by high-throughput sequencing, and circRNA-miRNA-mRNA network was constructed, so as to explore the function of non-coding RNA in the immune system of teleost. A total of 119, 140, and 510 differential expressed circRNAs, miRNAs, and mRNAs were identified in the infected groups compared with the uninfected group. The qRT-PCR verified the reliability and accuracy of the Illumina sequencing data. Fifteen triple networks of circRNA-miRNA-mRNA were presented in the form of “up (circRNA)-down (miRNA)-up (mRNA)” or “down-up-down”. Immune-related genes were also found in these networks. These results indicate that circRNAs and miRNAs may regulate the expression of immune-related genes through the circRNA-miRNA-mRNA regulatory network and thus participate in the immune response of turbot spleen after pathogen infection.Increasing evidence suggests that non-coding RNAs (ncRNA) play an important role in a variety of biological life processes by regulating gene expression at the transcriptional and post-transcriptional levels. Turbot (Scophthalmus maximus) has been threatened by various pathogens. In this study, the expression of circular RNAs (circRNAs), microRNAs (miRNAs), and mRNA in the immune organs spleen of turbot infected with Aeromonas salmonicida was analyzed by high-throughput sequencing, and a circRNA-miRNA-mRNA network was constructed, so as to explore the function of non-coding RNA in the immune system of teleost. Illumina sequencing was performed on the uninfected group and infected group. A total of 119 differential expressed circRNAs (DE-circRNAs), 140 DE-miRNAs, and 510 DE-mRNAs were identified in the four infected groups compared with the uninfected group. Most DE-mRNAs and the target genes of DE-ncRNAs were involved in immune-related pathways. The quantitative real-time PCR (qRT-PCR) results verified the reliability and accuracy of the high-throughput sequencing data. Ninety-six differentially expressed circRNA-miRNA-mRNA regulatory networks were finally constructed. Among them, 15 circRNA-miRNA-mRNA were presented in the form of “up (circRNA)-down (miRNA)-up (mRNA)” or “down-up-down”. Immune-related genes gap junction CX32.2, cell adhesion molecule 3, and CC chemokine were also found in these networks. These results indicate that ncRNA may regulate the expression of immune-related genes through the circRNA-miRNA-mRNA regulatory network and thus participate in the immune response of turbot spleen after pathogen infection.

Highlights

  • Turbot (Scophthalmus maximus, S. maximus) is a kind of cold-water benthic fish that originated from Europe, which is an important economic species in many countries

  • The adsorption of circRNAs to miRNAs prevents the binding of miRNA to its target mRNAs, indirectly regulating the expression of miRNA target genes [12]

  • We examined the expression of circRNA, miRNA, and mRNA in the spleen of turbot infected with A. salmonicida at different time points by high-throughput sequencing

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Summary

Introduction

Turbot (Scophthalmus maximus, S. maximus) is a kind of cold-water benthic fish that originated from Europe, which is an important economic species in many countries. With the development of next-generation sequencing technology and the integration analysis and application of multi-omics data, a variety of non-coding RNAs (ncRNAs), such as long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs), have been confirmed to be involved in the complex biological process of fish resistance to bacterial immunity. MicroRNA is a short (~22 nt), endogenous, evolutionarily conserved non-coding RNA that can down-regulate the expression of a target gene by binding to the 3 UTR of mature mRNA [11]. The adsorption of circRNAs to miRNAs prevents the binding of miRNA to its target mRNAs, indirectly regulating the expression of miRNA target genes [12]

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