Distinctive patterns of microRNA expression in extraocular muscles.
Distinctive patterns of microRNA expression in extraocular muscles.
复制标题
DOI：
10.1152/physiolgenomics.00169.2009
复制
发表时间：
2010-05
影响因子：
4.6
通讯作者：
U. Zeiger;T. Khurana
中科院分区：
文献类型：
--
作者：
U. Zeiger;T. Khurana
眼外肌(EOMS)是一组独特的肌肉，在解剖和生理上有别于其他肌肉。我们和其他人已经证明了EOMS具有独特的转录组和蛋白质组。在这里，我们研究了microRNAs(MiRNAs)的表达模式，因为它们可能在产生独特的EOM同种异型中发挥作用。我们分离了RNA并筛选了覆盖miRBase 10.0序列的LC Sciences miRNA微阵列，以确定正常小鼠EOM和胫骨前(TA)肢体肌肉的microRNAome。74个miRNAs被发现存在差异调控(P值500。肌特异性miR-206和miR-499在EOM中上调，miR-1、miR-133a和miR-133b下调。用定量聚合酶链式反应(QPCR)分析验证差异表达。生物信息学工具被用来确定潜在的miRNA-mRNA-蛋白质相互作用，并将数据与先前的转录组和蛋白质组学数据相结合。荧光素酶分析通过将前体miRNAs与包含预测目标基因的3‘-非翻译区的报告构建体共转染来验证已识别的miRNAs的靶向性。EOM microRNAome的定义补充了现有关于EOM分子组成的转录组和蛋白质组数据，并提供了对肌肉基因调控的进一步洞察。这些数据还将有助于进一步解释独特的EOM肌肉同种异型及其对Duchenne肌营养不良症等疾病的区分敏感性，并可能有助于制定治疗策略。
The extraocular muscles (EOMs) are a unique group of muscles that are anatomically and physiologically distinct from other muscles. We and others have shown that EOMs have a unique transcriptome and proteome. Here we investigated the expression pattern of microRNAs (miRNAs), as they may play a role in generating the unique EOM allotype. We isolated RNA and screened LC Sciences miRNA microarrays covering the sequences of miRBase 10.0 to define the microRNAome of normal mouse EOM and tibialis anterior (TA) limb muscle. Seventy-four miRNAs were found to be differentially regulated (P value 500. Muscle-specific miRNAs miR-206 and miR-499 were upregulated and miR-1, miR-133a, and miR-133b were downregulated in EOM. Quantitative PCR (qPCR) analysis was used to validate the differential expression. Bioinformatic tools were used to identify potential miRNA-mRNA-protein interactions and integrate data with previous transcriptome and proteomic profiling data. Luciferase assays using cotransfection of precursor miRNAs with reporter constructs containing the 3'-untranslated region of predicted target genes were used to validate targeting by identified miRNAs. The definition of the EOM microRNAome complements existing transcriptome and proteome data about the molecular makeup of EOM and provides further insight into regulation of muscle genes. These data will also help to further explain the unique EOM muscle allotype and its differential sensitivity to diseases such as Duchenne muscular dystrophy and may assist in development of therapeutic strategies.