Abstract
BackgroundMany recent studies have shown that miRNAs play important roles in the regulation of animal reproduction, including seasonal reproduction. The pineal gland is a crucial hub in the regulation of seasonal reproduction. However, little is known about the expression characteristics of pineal miRNAs in different reproductive seasons (anestrus and breeding season). Therefore, the expression profiles and regulatory roles of ovine pineal miRNAs were investigated during different reproductive stages using Solexa sequencing technology and dual luciferase reporter assays.ResultsA total of 427 miRNAs were identified in the sheep pineal gland. Significant differences in miRNA expression were demonstrated between anestrus and the breeding season in terms of the frequency distributions of miRNA lengths, number of expressed miRNAs, and specifically and highly expressed miRNAs in each reproductive stage. KEGG analysis of the differentially expressed (DE) miRNAs between anestrus and the breeding season indicated that they are significantly enriched in pathways related to protein synthesis, secretion and uptake. Furthermore, transcriptome analysis revealed that many target genes of DE miRNAs in the ribosome pathway showed relatively low expression in the breeding season. On the other hand, analyses combining miRNA-gene expression data with target relationship validation in vitro implied that miR-89 may participate in the negative regulation of aralkylamine N-acetyltransferase (AANAT) mRNA expression by targeting its 3’UTR at a unique binding site.ConclusionsOur results provide new insights into the expression characteristics of sheep pineal miRNAs at different reproductive stages and into the negative regulatory effects of pineal miRNAs on AANAT mRNA expression.
Highlights
Many recent studies have shown that miRNAs play important roles in the regulation of animal reproduction, including seasonal reproduction
Expression characteristics of pineal small RNAs in sheep at different reproductive stages Pineal tissues were collected at different reproductive stages, and the accuracy of the sampling stage determination was confirmed by the results obtained from ovarian sections (Fig. 1)
To further assess the efficiency of highthroughput sequencing in detecting miRNA, all the mapped clean reads were annotated and classified by alignment against noncoding RNAs (ncRNAs)
Summary
Many recent studies have shown that miRNAs play important roles in the regulation of animal reproduction, including seasonal reproduction. Little is known about the expression characteristics of pineal miRNAs in different reproductive seasons (anestrus and breeding season). Many studies have shown that miRNAs play important regulatory roles in animal reproduction [3,4,5,6,7,8,9]. Little is known about the expression characteristics of pineal miRNAs in different reproductive seasons (nonbreeding and breeding seasons). One objective of this study was to reveal the expression characteristics of pineal miRNAs in sheep during different reproductive stages. The second purpose of this study was to screen differentially expressed miRNAs among the different reproductive stages and predict their probable functions in the pineal gland
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