The identification and characteristics of immune-related microRNAs in haemocytes of oyster Crassostrea gigas

PLoS One. 2014 Feb 6;9(2):e88397. doi: 10.1371/journal.pone.0088397. eCollection 2014.

Abstract

Background: MicroRNAs (miRNAs) represent a class of small ncRNAs that repress gene expression on the post-transcriptional level by the degradation or translation inhibition of target mRNA.

Methodology: Three small RNA libraries from oyster haemocytes were sequenced on the Illumina platform to investigate the latent immunomodulation of miRNAs after bacteria challenge and heat stress. Totally, 10,498,663, 8,588,606 and 9,679,663 high-quality reads were obtained in the control, bacteria and bacteria+heat library, respectively, from which 199 oyster miRNAs including 71 known and 128 novel ones were identified. Among these miRNAs, 6 known and 23 novel ones were predicted to possess more than one precursor-coding region, and cgi-miR-10a, cgi-miR-184b, cgi-miR-100, cgi-miR-1984 and cgi-miR-67a were observed to be the most abundant miRNAs in the control library. The expression levels of 22 miRNAs in the bacteria library were significantly higher than those in the control library, while there were another 33 miRNAs whose expression levels were significantly lower than that in the control library. Meanwhile, the expression levels of 65 miRNAs in the bacteria+heat library changed significantly compared to those in the bacteria library. The target genes of these differentially expressed miRNAs were annotated, and they fell in immune and stress-related GO terms including antioxidant, cell killing, death, immune system process, and response to stimulus. Furthermore, there were 42 differentially expressed miRNAs detected in both control/bacteria and bacteria/bacteria+heat comparisons, among which 9 miRNAs displayed the identical pattern in the two comparisons, and the expression alterations of 8 miRNAs were confirmed using quantitative RT-PCR.

Conclusions: These results indicated collectively that immune challenge could induce the expression of immune-related miRNAs, which might modulate the immune response such as redox reaction, phagocytosis and apoptosis, and the expression of some immune-related miRNAs could be also regulated by heat stress to improve the environmental adaption of oyster.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cluster Analysis
  • Crassostrea / cytology
  • Crassostrea / genetics*
  • Crassostrea / immunology*
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Gene Library
  • Gene Ontology
  • Heat-Shock Response / genetics
  • Hemocytes / metabolism*
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Molecular Sequence Data
  • Open Reading Frames / genetics
  • Real-Time Polymerase Chain Reaction
  • Reproducibility of Results
  • Sequence Analysis, RNA

Substances

  • MicroRNAs

Grants and funding

This research was supported by 973 National Key Fundamental Research Program (No. 2010CB126404), National High Technology Research and Development Program (863 Program, No. 2012AA10A401) from the Chinese Ministry of Science and Technology (http://www.most.gov.cn/), grants from NSFC (No. 31072192 to L.W, 30925028 to L.S, http://www.nsfc. gov.cn/), and Shandong Provincial Natural Science Foundation (No. JQ201110 to L.W, http://www.sdnsf.gov.cn/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.