RT-PCR-based analysis of microRNA (miR-1 and-124) expression in mouse CNS
RT-PCR-based analysis of microRNA (miR-1 and-124) expression in mouse CNS
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DOI：
10.1016/j.brainres.2006.11.035
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发表时间：
2007-02-02
期刊：
影响因子：
2.9
通讯作者：
Takizawa, Toshihiro
中科院分区：
文献类型：
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作者：
Mishima, Takuya;Mizuguchi, Yoshiaki;Takizawa, Toshihiro
超过700种microrna （mirna）已经被克隆，这些分子在发育定时、细胞增殖和癌症中的功能已经被广泛研究。用Northern blot和序列集落评价分析mirna；然而，基于逆转录聚合酶链反应（RT-PCR）的miRNA检测仍有待开发。在本报告中，我们描述了改进的实时RT- pcr方法，使用特异性或非特异性RT引物进行miRNA表达的半定量分析。在模型研究中使用的新方法揭示了miRNA-1 （miR-1）和miR-124在小鼠器官中的差异表达。具体来说，我们的方法揭示了miR-124在小鼠中枢神经系统（CNS，大脑皮层，小脑和脊髓）中的浓度是其他器官中的浓度的100多倍。相比之下，miR-1在中枢神经系统中的表达比骨骼肌和心脏低100-1000倍。此外，我们揭示了中枢神经系统内miR-124表达的解剖区域差异：小脑与大脑皮层的表达率为60.7%，脊髓为35.4%。这些结果表明，我们基于rt - pcr的方法将成为研究与各种神经事件相关的miRNA表达的有力工具。(c) 2006 Elsevier B.V.版权所有
More than 700 microRNAs (miRNAs) have been cloned, and the functions of these molecules in developmental timing, cell proliferation, and cancer have been investigated widely. MiRNAs are analyzed with Northern blot and sequential colony evaluation; however, reverse transcription-polymerase chain reaction (RT-PCR)-based miRNA assay remains to be developed. In this report, we describe improved real-time RT-PCR methods using specific or non-specific RT primer for the semi-quantitative analysis of miRNA expression. The use of the new methods in a model study revealed differential expression of miRNA-1 (miR-1) and miR-124 in mouse organs. Specifically, our methods revealed that miR-124 concentrations in the mouse central nervous system (CNS; cerebral cortex, cerebellum, and spinal cord) were more than 100 times those in other organs. By contrast, miR-1 expression in the CNS was 100-1000 times lower than that in skeletal muscle and heart. Furthermore, we revealed anatomically regional differences in miR-124 expression within the CNS: expression ratios versus the cerebral cortex were 60.7% for the cerebellum and 35.4% for the spinal cord. These results suggest that our RT-PCR-based methods would be a powerful tool for studies of miRNA expression that is associated with various neural events. (c) 2006 Elsevier B.V. All rights reserved.