Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian, P.R. China.
PLoS One. 2012;7(11):e49463. doi: 10.1371/journal.pone.0049463. Epub 2012 Nov 14.
MicroRNAs are small, noncoding RNA molecules that regulate gene expression at the post-transcriptional level and play an important role in various biological processes. Although most microRNAs expression profiles studies have been performed in humans or rodents, relatively limited knowledge also exists in other mammalian species. The identification of the full repertoire of microRNAs expressed in the lactating mammary gland of Capra hircus would significantly increase our understanding of the physiology of lactating mammary glands. In this study, two libraries were constructed using the lactating mammary gland tissues of Laoshan dairy goats (Capra hircus) during peak and late lactation. Solexa high-throughput sequencing technique and bioinformatics were used to determine the abundance and differential expression of the microRNAs between peak and late lactation. As a result, 19,044,002 and 7,385,833 clean reads were obtained, respectively, and 1,113 conserved known microRNAs and 31 potential novel microRNA candidates were identified. A total of 697 conserved microRNAs were significantly differentially expressed with a P-value<0.01, 272 microRNAs were up-regulated and 425 microRNAs were down-regulated during peak lactation. The results were validated using real-time quantitative RT-PCR. 762,557 annotated mRNA transcripts were predicted as putative target gene candidates. The GO annotation and KEGG pathway analysis suggested that differentially expressed microRNAs were involved in mammary gland physiology, including signal transduction, and cell-cell and cell-extracellular communications. This study provided the first global of the microRNA in Capra hircus and expanded the repertoire of microRNAs. Our results have great significance and value for the elucidation of complex regulatory networks between microRNAs and mRNAs and for the study of mammary gland physiology and lactation.
microRNAs 是一类小的非编码 RNA 分子,可在转录后水平调控基因表达,在多种生物学过程中发挥重要作用。尽管大多数 microRNAs 表达谱研究都是在人类或啮齿动物中进行的,但在其他哺乳动物物种中也有相对有限的知识。鉴定绵羊乳腺中表达的完整 microRNA 谱将显著提高我们对泌乳乳腺生理学的理解。在这项研究中,使用崂山奶山羊(Capra hircus)泌乳期乳腺组织构建了两个文库。采用 Solexa 高通量测序技术和生物信息学方法,确定了泌乳高峰期和泌乳后期乳腺组织中 microRNAs 的丰度和差异表达。结果分别获得了 19,044,002 和 7,385,833 条清洁读段,鉴定出 1,113 个保守的已知 microRNAs 和 31 个潜在的新 microRNA 候选物。共有 697 个保守 microRNAs 的差异表达具有统计学意义(P 值<0.01),其中 272 个 microRNAs 在泌乳高峰期上调,425 个 microRNAs 下调。采用实时定量 RT-PCR 对结果进行验证。预测了 762,557 个注释的 mRNA 转录本作为潜在的靶基因候选物。GO 注释和 KEGG 通路分析表明,差异表达的 microRNAs参与了乳腺生理学,包括信号转导以及细胞-细胞和细胞-细胞外通讯。本研究提供了绵羊 Capra hircus 中 microRNA 的首个全景图,并扩展了 microRNA 谱。我们的研究结果对于阐明 microRNA 与 mRNA 之间的复杂调控网络以及研究乳腺生理学和泌乳具有重要意义和价值。