Microarray-Based Approach Identifies Differentially Expressed MicroRNAs in Porcine Sexually Immature and Mature Testes

Lifan Luo,Siwen Jiang,Fenge Li,Bo Zuo,Lianzhi Ye,Dequan Xu,Guochao Shao,Changyan Deng,Yuanzhu Xiong,Minggang Lei,Zhuqing Ren,Rong Zheng,Gang Liu,Alan Christoffels

doi:10.1371/journal.pone.0011744

Abstract

BackgroundMicroRNAs (miRNAs) are short non-coding RNA molecules which are proved to be involved in mammalian spermatogenesis. Their expression and function in the porcine germ cells are not fully understood.MethodologyWe employed a miRNA microarray containing 1260 unique miRNA probes to evaluate the miRNA expression patterns between sexually immature (60-day) and mature (180-day) pig testes. One hundred and twenty nine miRNAs representing 164 reporter miRNAs were expressed differently (p<0.1). Fifty one miRNAs were significantly up-regulated and 78 miRNAs were down-regulated in mature testes. Nine of these differentially expressed miRNAs were validated using quantitative RT-PCR assay. Totally 15919 putative miRNA-target sites were detected by using RNA22 method to align 445 NCBI pig cDNA sequences with these 129 differentially expressed miRNAs, and seven putative target genes involved in spermatogenesis including DAZL, RNF4 gene were simply confirmed by quantitative RT-PCR.ConclusionsOverall, the results of this study indicated specific miRNAs expression in porcine testes and suggested that miRNAs had a role in regulating spermatogenesis.

Highlights

MicroRNAs are small non-coding RNAs that play important roles in regulating posttranscriptional translation
Overall, the results of this study indicated specific miRNAs expression in porcine testes and suggested that miRNAs had a role in regulating spermatogenesis
10883 miRNA sequences have been published on the Sanger miRNA Registry. They are increasingly being shown to play vital roles in spermatogenesis, muscle development, feed intake and other important physiological process [2,3,4]. Such as the myostatin allele in muscle mass QTL interval is characterized by a G to A transition in the 39 un-translated region (UTR) that creates a target site for miR-1 and miR-206 which are highly expressed in skeletal muscle

Summary

Introduction

MicroRNAs (miRNAs) are small non-coding RNAs (typically 19–23 nucleotides) that play important roles in regulating posttranscriptional translation. MiR-122a was suggested targeting a reporter mRNA containing sequences from the 39-UTR of the transition protein 2 (TNP2), a post-transcriptionally regulated testis-specific gene involved in chromatin remodeling during mouse spermatogenesis [2].

Results

Conclusion