Differential promoter usage of mouse μ-opioid receptor gene during development
Differential promoter usage of mouse μ-opioid receptor gene during development
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DOI：
10.1016/s0169-328x(02)00357-1
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发表时间：
2002-08-15
期刊：
影响因子：
--
通讯作者：
Loh, HH
中科院分区：
文献类型：
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作者：
Ko, JL;Chen, HC;Loh, HH
先前，我们证明小鼠mu-阿片受体（MOR）基因表达受远端启动子和近端启动子调控，其中近端启动子在成年小鼠大脑中控制MOR转录起主要作用。在这里，我们报道了小鼠MOR双启动子在小鼠发育过程中的相关用法的研究。我们使用逆转录聚合酶链反应（RT-PCR）方法，其结果与使用结合试验或原位杂交的结果相似。然而，由于RT-PCR方法的灵敏度更高，我们能够在胚胎第8.5天检测到MOR的出现（E8.5）。我们发现在E8.5时，近端启动子和远端启动子都是活跃的。在E8.5中，近端启动子启动了大约三分之二的MOR转录本，远端启动子指导剩余三分之一的转录。这是我们在发育过程中观察到的远端启动子对MOR转录的最大相对贡献。此后，由远端启动子引导的转录本百分比逐渐下降，并在整个发育和成年期间保持在低但可检测的水平（约占总MOR转录本的5%）。相反，在发育过程中观察到近端启动子对MOR转录的贡献逐渐增加，在成人中达到最大。总之，我们的研究结果证明了近端启动子在小鼠发育过程中指导MOR转录的关键作用。(C) 2002 Elsevier Science B.V.版权所有
Previously, we demonstrated that mouse mu-opioid receptor (MOR) gene expression is regulated by both distal and proximal promoters, with the latter playing a major role in controlling MOR transcription in the adult mouse brain. Here, we report studies of the relative usages of the mouse MOR dual promoters during murine development. We used the reverse transcription-polymerase chain reaction (RT-PCR) method, which gave results similar to those using binding assays or in situ hybridization. However, due to the greater sensitivity of RT-PCR method, we were able to detect the emergence of MOR as early as at embryonic day 8.5 (E8.5). We found that both proximal and distal promoters were active at E8.5. The proximal promoter initiated approximately two-thirds of total MOR transcripts at E8.5, with the distal promoter directing transcription of the remaining one-third. This is the greatest relative contribution of the distal promoter to MOR transcription we have observed during any time in development. Thereafter, the percentage of transcripts directed by the distal promoter gradually declined, and remained at a low but detectable level (approximately 5% of total MOR transcripts) throughout development and adulthood. Conversely, a progressive increase of the contribution of the proximal promoter to MOR transcription was observed during development, reaching its maximum in the adult. In summary, our results demonstrated the pivotal role of the proximal promoter in directing MOR transcription during murine development. (C) 2002 Elsevier Science B.V. All rights reserved.