In vivo and in vitro visualization of gene expression dynamics over extensive areas of the brain
In vivo and in vitro visualization of gene expression dynamics over extensive areas of the brain
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DOI：
10.1016/j.neuroimage.2008.10.046
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发表时间：
2009-02-15
期刊：
影响因子：
5.7
通讯作者：
Yamaguchi, Shun
中科院分区：
文献类型：
--
作者：
Eguchi, Megumi;Yamaguchi, Shun
使用启动子不稳定荧光蛋白构建体在体内监测基因表达是检测大脑中即时早期基因表达动态的有力方法。然而，这种结构产生的微弱荧光信号阻碍了对大脑广泛区域基因表达的分析。我们成功地培育出了高水平表达Arc基因启动子驱动的报告基因的转基因小鼠，该基因的激活与多种脑功能相关（报告mRNA丰度比内源性Arc mRNA高近100倍）。报告基因的高表达使我们能够监测Arc基因在体内的表达动态，包括整个大脑背皮层。此外，我们能够使用多聚甲醛固定的大脑对激活区域进行三维分析。除了视觉皮层，我们发现扣带皮层被光刺激强烈激活。这些小鼠对于野生型小鼠和大脑功能受损的突变体的大脑广泛区域的基因表达功能分析非常有用。(C) 2008爱思唯尔公司版权所有。
In vivo monitoring of gene expression using promoter-destabilized fluorescence protein constructs is a powerful method for examining the expression dynamics of immediate-early genes in the brain. However, weak fluorescence signals derived from such constructs have hampered analyses of gene expression over extensive areas of the brain. We succeeded in producing transgenic mice with brains exhibiting high level expression of the reporter gene driven by the Arc gene promoter, which is activated in association with various brain functions (reporter mRNA abundance was near 100-fold greater than endogenous Arc mRNAs). This high expression of the reporter gene enabled us to monitor Arc gene expression dynamics in vivo, over an area that included the whole of the dorsal cerebral cortex. Moreover, we were able to perform three-dimensional analyses of activated regions using paraformaldehyde-fixed brains. In addition to the visual cortex, we found that the cingulate cortex was strongly activated by light stimuli. These mice are extremely useful for the functional analysis of gene expression over extensive areas of the brains in both wild-type mice and mutants with impaired brain function. (C) 2008 Elsevier Inc. All rights reserved.