miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family.
miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family.
复制标题
DOI：
10.1261/rna.034066.112
复制
发表时间：
2014-12
期刊：
影响因子：
--
通讯作者：
Gardner PD
中科院分区：
文献类型：
--
作者：
Hogan EM;Casserly AP;Scofield MD;Mou Z;Zhao-Shea R;Johnson CW;Tapper AR;Gardner PD
尼古丁结合并激活配体门控离子通道家族，神经元烟碱乙酰胆碱受体（nAChR）。慢性尼古丁暴露改变了各种nAChR亚型的表达，这可能有助于尼古丁依赖;然而，调节这些变化的潜在机制仍不清楚。越来越多的证据表明microRNAs（miRNAs）可能参与nAChR的调控。利用生物信息学、miRNA文库筛选、定点突变和基因表达分析等方法，我们鉴定了少数与哺乳动物神经元nAChR亚基基因3′非翻译区（3′ UTRs）功能性相互作用的miRNA。计算机模拟分析揭示了3′ UTR中特定的、进化上保守的位点，miRNA通过这些位点调节基因表达。这些位点的突变破坏了miRNA的调控，证实了计算机模拟的预测。此外，靶向nAChR 3′ UTR的miRNA在小鼠脑中表达，并受慢性尼古丁暴露的调节。此外，我们还表明，其中一种miRNA（miR-542- 3 p）的表达受到中皮质边缘奖励途径中尼古丁的调节。重要的是，miR-542- 3 p的过表达导致其靶点nAChR β2亚基的蛋白水平降低。生物信息学分析表明，许多miRNAs在调节胆碱能信号中起着普遍的作用。我们的研究结果提供了一种新的尼古丁介导的哺乳动物nAChR基因家族的调节模式的证据。
Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3′-untranslated regions (3′ UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3′ UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3′ UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR β2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family.
登录
查看更多内容
影响因子：
25
作者：
Im, Heh-In;Hollander, Jonathan A.;Bali, Purva;Kenny, Paul J.
通讯作者：
Kenny, Paul J.
哺乳动物烟碱乙酰胆碱受体：从结构到功能。
影响因子：
33.6
作者：
Albuquerque EX;Pereira EF;Alkondon M;Rogers SW
通讯作者：
Rogers SW
Mirbase：MicroRNA基因组学的工具。
影响因子：
14.9
作者：
Griffiths-Jones, Sam;Saini, Harpreet Kaur;van Dongen, Stijn;Enright, Anton J.
通讯作者：
Enright, Anton J.
影响因子：
4.7
作者：
BENWELL, MEM;BALFOUR, DJK;ANDERSON, JM
通讯作者：
ANDERSON, JM
影响因子：
4.8
作者：
Ficklin, MB;Zhao, SL;Feng, GP
通讯作者：
Feng, GP