Prediction of microRNAs affecting mRNA expression during retinal development.
Prediction of microRNAs affecting mRNA expression during retinal development.
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DOI：
10.1186/1471-213x-10-1
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发表时间：
2010-01-06
影响因子：
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通讯作者：
Simpson DA
中科院分区：
文献类型：
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作者：
Arora A;Guduric-Fuchs J;Harwood L;Dellett M;Cogliati T;Simpson DA
MicroRNA (miRNA) 是小 RNA 分子（约 22 个核苷酸），已被证明在成体组织的发育和维护中发挥着重要作用。眼睛中 miRNA 的条件性失活会导致视觉功能丧失和进行性视网膜变性。除了抑制翻译之外，miRNA 还可以介导目标 mRNA 的降解。我们之前已经表明，影响组织中转录水平的候选 miRNA 可以从 mRNA 微阵列表达谱中推断出来。本研究的目的是预测影响发育中和成年视网膜组织中 mRNA 水平的 miRNA，并确认其表达。睫状上皮视网膜干细胞（CE-RSC）、发育中和成年小鼠视网膜的微阵列表达数据是从公共存储库生成或下载的。基因表达谱分析检测了多种 miRNA 在 CE-RSC 和视网膜中的作用。通过 RT-PCR 确认了 20 个选定 miRNA 的表达，并通过原位杂交分析了代表性候选者的细胞分布。 miRNA 的表达水平与其对 mRNA 表达的预测影响的显着性相关。高表达的 miRNA 包括 miR-124、miR-125a、miR-125b、miR-204 和 miR-9。三种 miRNA 的过度表达对整体 mRNA 水平具有显着的预测影响，导致所分析的 6 个单独预测靶基因中的 5 个 mRNA 表达下降。本研究检测了 miRNA 对未成熟和成年视网膜组织和细胞中 mRNA 表达的影响。这些观察结果的有效性得到了候选 miRNA 表达的实验证实以及 miRNA 过度表达后预测靶基因的调节。已鉴定的 miRNA 可能对视网膜发育和功能很重要。这些 miRNA 的失调可能会导致视网膜变性和疾病。相反，操纵它们的表达可能会在未来用作治疗工具。
MicroRNAs (miRNAs) are small RNA molecules (~22 nucleotides) which have been shown to play an important role both in development and in maintenance of adult tissue. Conditional inactivation of miRNAs in the eye causes loss of visual function and progressive retinal degeneration. In addition to inhibiting translation, miRNAs can mediate degradation of targeted mRNAs. We have previously shown that candidate miRNAs affecting transcript levels in a tissue can be deduced from mRNA microarray expression profiles. The purpose of this study was to predict miRNAs which affect mRNA levels in developing and adult retinal tissue and to confirm their expression. Microarray expression data from ciliary epithelial retinal stem cells (CE-RSCs), developing and adult mouse retina were generated or downloaded from public repositories. Analysis of gene expression profiles detected the effects of multiple miRNAs in CE-RSCs and retina. The expression of 20 selected miRNAs was confirmed by RT-PCR and the cellular distribution of representative candidates analyzed by in situ hybridization. The expression levels of miRNAs correlated with the significance of their predicted effects upon mRNA expression. Highly expressed miRNAs included miR-124, miR-125a, miR-125b, miR-204 and miR-9. Over-expression of three miRNAs with significant predicted effects upon global mRNA levels resulted in a decrease in mRNA expression of five out of six individual predicted target genes assayed. This study has detected the effect of miRNAs upon mRNA expression in immature and adult retinal tissue and cells. The validity of these observations is supported by the experimental confirmation of candidate miRNA expression and the regulation of predicted target genes following miRNA over-expression. Identified miRNAs are likely to be important in retinal development and function. Misregulation of these miRNAs might contribute to retinal degeneration and disease. Conversely, manipulation of their expression could potentially be used as a therapeutic tool in the future.