Synaptic activity-responsive element in the Arc/Arg3.1 promoter essential for synapse-to-nucleus signaling in activated neurons
Synaptic activity-responsive element in the Arc/Arg3.1 promoter essential for synapse-to-nucleus signaling in activated neurons
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DOI：
10.1073/pnas.0806518106
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发表时间：
2009-01-06
影响因子：
11.1
通讯作者：
Bito, Haruhiko
中科院分区：
文献类型：
--
作者：
Kawashima, Takashi;Okuno, Hiroyuki;Bito, Haruhiko
神经元即早基因Arc/Arg-3.1被广泛用作体内强烈突触活动的最可靠的分子标记之一。然而，负责这种严格的活性依赖性的顺式作用元件还没有被确定。在这里，我们结合荧光素酶报告基因检测培养的皮层神经元和比较基因组定位，以确定关键的突触活动响应元件（萨雷）的弧/精氨酸-3.1基因。发现一个主要的萨雷作为一个独特的约100 bp的元件，位于小鼠基因组中Arc/Arg-3.1转录起始位点上游>5 kb处。这个单一的元素，当定位在一个最小的启动子的上游，是必要的和足够的复制内源性弧/精氨酸-3.1的转录调控的关键特性，包括快速启动的转录触发突触活动和低基础表达在突触不活动。我们确定了萨雷的主要决定因素，作为一个独特的神经元活性依赖性顺式调节元件的集群组成的密切本地化的结合位点CREB，MEF 2，和SRF。因此，萨雷报告基因可以容易地追踪和标记最近在体内经历过强烈活性的细胞系。总之，我们的工作揭示了一种新的转录机制，其中一个关键的100 bp的元件，萨雷，介导了一个主要组成部分的突触到核信号的合奏弧/精氨酸-3.1阳性激活的神经元。
The neuronal immediate early gene Arc/Arg-3.1 is widely used as one of the most reliable molecular markers for intense synaptic activity in vivo. However, the cis-acting elements responsible for such stringent activity dependence have not been firmly identified. Here we combined luciferase reporter assays in cultured cortical neurons and comparative genome mapping to identify the critical synaptic activity-responsive elements (SARE) of the Arc/Arg-3.1 gene. A major SARE was found as a unique approximate to 100-bp element located at >5 kb upstream of the Arc/Arg-3.1 transcription initiation site in the mouse genome. This single element, when positioned immediately upstream of a minimal promoter, was necessary and sufficient to replicate crucial properties of endogenous Arc/Arg-3.1's transcriptional regulation, including rapid onset of transcription triggered by synaptic activity and low basal expression during synaptic inactivity. We identified the major determinants of SARE as a unique cluster of neuronal activity-dependent cis-regulatory elements consisting of closely localized binding sites for CREB, MEF2, and SRF. Consistently, a SARE reporter could readily trace and mark an ensemble of cells that have experienced intense activity in the recent past in vivo. Taken together, our work uncovers a novel transcriptional mechanism by which a critical 100-bp element, SARE, mediates a predominant component of the synapse-to-nucleus signaling in ensembles of Arc/Arg-3.1-positive activated neurons.