期刊论文详细信息
BMC Genomics
Global analysis of the ovarian microRNA transcriptome: implication for miR-2 and miR-133 regulation of oocyte meiosis in the Chinese mitten crab, Eriocheir sinensis (Crustacea:Decapoda)
Gao-Feng Qiu1  Zhi-Qiang Liu1  Li-Li Shi1  Ya-Nan Song1 
[1] Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Certificated by Ministry of Education, College of Fisheries and Life Science, Shanghai Ocean University, 999 Hucheng Huan Road, Pudong New Area, Shanghai 201306, China
关键词: Chinese mitten crab;    Oocyte meiosis;    MicroRNA transcriptome;    Ovary;   
Others  :  856679
DOI  :  10.1186/1471-2164-15-547
 received in 2014-04-14, accepted in 2014-06-24,  发布年份 2014
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【 摘 要 】

Background

MicroRNAs (miRNAs) are small non-coding RNA molecules that downregulate gene expression by base pairing to the 3′-untranslated region (UTR) of target messenger RNAs (mRNAs). Up to now, rare information for the miRNAs is available in decapod crustaceans. Our previous studies showed that many miRNA-binding sites are present in the 3′-UTR of the cyclin B in the Chinese mitten crab Eriocheir sinensis, suggesting that the translation or post-transcription of the crab cyclin B might be regulated by miRNAs during meiosis of oocyte.

Results

To identify ovarian miRNAs in the mitten crab, ovarian small RNAs were subjected to high-throughput sequencing using an Illumina Genome Analyzer. Of 14,631,328 reads, 55 known miRNAs representing 44 miRNA families were identified and 136 novel miRNA candidates were predicted. The 5′ seed sequences of four miRNAs, miR-2, miR-7, miR-79 and miR-133, were revealed to complementary to miRNA binding sites in 3′-UTR of the cyclin B. Quantitative real time PCR analysis showed that miR-2 and miR-133 are much more abundant in the first metaphase (MI) of meiosis than in germinal vesicle (GV) stage. But their increasing expressions are independent of induction of gonadotropin-releasing hormone (GnRH). Further expression analysis using double-luciferase reporter genes assay showed that miR-2 and miR-133 can downregulate the 3′-UTRs of the crab cyclin B gene, indicating that they could inhibit the translation of the cyclin B. Western blot analysis confirmed that cyclin B protein is completely disappeared in fertilized egg at the metaphase-anaphase transition of meiosis I, suggesting that miR-2 and miR-133 could function in destruction of cyclin B near the end of MI.

Conclusions

A high number of miRNAs have been identified from the crab ovarian small RNA transcriptom for the first time. miR-2 and miR-133 exhibit differential expression during the meiotic maturation of the oocytes and have activity in regulating the 3′-UTR of the crab cyclin B gene. This result is inconsistent with recent finding that miRNA activity is globally suppressed in mouse oocytes.

【 授权许可】

   
2014 Song et al.; licensee BioMed Central Ltd.

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