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:DecapodaReportar como inadecuado




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 - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Genomics

, 15:547

Multicellular invertebrate genomics

Abstract

BackgroundMicroRNAs 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.

ResultsTo 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.

ConclusionsA 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.

KeywordsOvary MicroRNA transcriptome Oocyte meiosis Chinese mitten crab AbbreviationsMiRNAMicroRNA

siRNASmall interfering RNA

piRNAPiwi-associatedRNA

UTRsUntranslated regions

Dgcr8DiGeorge syndrome critical region 8

RISCRNA-induced silencing complex

ntNucleotides

ESTExpressed sequence tag

GnRHGonadotropin-releasing hormone

MPFMaturation-promoting factor

CPECytoplasmic polyadenylation element

TCETranslation controlelement

PBSPhosphate buffered saline

GVGerminal vesicle

GVBDGerminal vesicle breakdown.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2164-15-547 contains supplementary material, which is available to authorized users.

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Autor: Ya-Nan Song - Li-Li Shi - Zhi-Qiang Liu - Gao-Feng Qiu

Fuente: https://link.springer.com/



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