NMDA receptor-dependent regulation of miRNA expression and association with Argonaute during LTP in vivo.
NMDA receptor-dependent regulation of miRNA expression and association with Argonaute during LTP in vivo.
复制标题
DOI：
10.3389/fncel.2013.00285
复制
发表时间：
2014-01-13
影响因子：
5.3
通讯作者：
Bramham CR
中科院分区：
文献类型：
--
作者：
Pai B;Siripornmongcolchai T;Berentsen B;Pakzad A;Vieuille C;Pallesen S;Pajak M;Simpson TI;Armstrong JD;Wibrand K;Bramham CR
microRNA（miRNAs）是大脑中蛋白质合成的主要调节因子。一个主要的目标是确定潜在的蛋白质合成依赖形式的突触可塑性，如长时程增强（LTP）的miRNA表达的变化。以前的分析集中在总裂解物样品中miRNA水平的变化。在这里，我们询问总miRNA的变化是否准确地反映了miRNA诱导的沉默复合物（miRISC）中与Argonaute蛋白结合的miRNA量的变化。Ago 2免疫沉淀用于分离麻醉大鼠齿状回中高频刺激（HFS）诱导的LTP后RISC相关的miRNA。使用基于锁核酸的PCR卡进行高通量筛选，并通过定量TaqMan RT-PCR进行独立验证，我们确定了Ago 2相关和总miRNA表达的差异调节。Ago 2/总miRNA表达的比例以miRNA特异性方式双向调节，并且在很大程度上依赖于LTP诱导期间N-甲基-D-天冬氨酸受体（NMDA）的激活。目前的结果确定了miRNA与Ago 2的关联作为成人大脑中活动依赖性突触可塑性的潜在控制点。最后，新的计算分析Ago 2相关的miRNAs的目标，确定了21个途径，富集和差异靶向的miRNAs，包括轴突导向，mTOR，MAPK，Ras，和LTP。
microRNAs (miRNAs) are major regulators of protein synthesis in the brain. A major goal is to identify changes in miRNA expression underlying protein synthesis-dependent forms of synaptic plasticity such as long-term potentiation (LTP). Previous analyses focused on changes in miRNA levels in total lysate samples. Here, we asked whether changes in total miRNA accurately reflect changes in the amount of miRNA bound to Argonaute protein within the miRNA-induced silencing complex (miRISC). Ago2 immunoprecipitation was used to isolate RISC-associated miRNAs following high-frequency stimulation (HFS)-induced LTP in the dentate gyrus of anesthetized rats. Using locked-nucleic acid-based PCR cards for high-throughput screening and independent validation by quantitative TaqMan RT-PCR, we identified differential regulation of Ago2-associated and total miRNA expression. The ratio of Ago2/total miRNA expression was regulated bidirectionally in a miRNA-specific manner and was largely dependent on N-methyl-D-aspartate receptor (NMDA) activation during LTP induction. The present results identify miRNA association with Ago2 as a potential control point in activity-dependent synaptic plasticity in the adult brain. Finally, novel computational analysis for targets of the Ago2-associated miRNAs identifies 21 pathways that are enriched and differentially targeted by the miRNAs including axon guidance, mTOR, MAPK, Ras, and LTP.
登录
查看更多内容
影响因子：
64.5
作者：
Ashraf, SI;McLoon, AL;Kunes, S
通讯作者：
Kunes, S
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：
64.5
作者：
Helwak A;Kudla G;Dudnakova T;Tollervey D
通讯作者：
Tollervey D
影响因子：
15.9
作者：
Hoeffer, Charles A.;Klann, Eric
通讯作者：
Klann, Eric
影响因子：
8.8
作者：
Horn, Katherine E.;Glasgow, Stephen D.;Kennedy, Timothy E.
通讯作者：
Kennedy, Timothy E.