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通过文库表达差异分析(DAzLE)鉴定钙和一氧化氮调节的基因。

Identification of calcium- and nitric oxide-regulated genes by differential analysis of library expression (DAzLE).

作者信息

Li Huiwu, Gu Xiujing, Dawson Valina L, Dawson Ted M

机构信息

Institute for Cell Engineering and Department of Neurology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

出版信息

Proc Natl Acad Sci U S A. 2004 Jan 13;101(2):647-52. doi: 10.1073/pnas.0305145101. Epub 2003 Dec 30.

Abstract

Using a method of expression profiling called differential analysis of cDNA library expression (DAzLE), we report the expression profile of late response genes in a model of activity-dependent neuronal survival and neurite outgrowth. Using DAzLE, we isolated differentially expressed genes from cultured rat embryonic cortical neurons after KCl (50 mM)-mediated membrane depolarization. We identified 469 activity-dependent regulated genes, of which 174 are genes of unknown function. The regulation of 63 genes was found to be nitric oxide (NO)-dependent. Identifiable genes fell into several major categories, including signal transduction pathways, neuronal development, DNA replication, gene transcription, protein metabolism, energy regulatory proteins, and antiapoptotic proteins. These genes may be important in activity-dependent neuron survival and development. Furthermore, these late response genes provide the tools to begin to investigate downstream events in activity-dependent neuronal survival and development. The major advantage of DAzLE is that it provides a nearly complete and relatively comprehensive differential screening profile that has the potential to be a powerful and useful tool in other fields of study.

摘要

我们使用一种名为cDNA文库表达差异分析(DAzLE)的表达谱分析方法,报告了在活动依赖型神经元存活和神经突生长模型中晚期反应基因的表达谱。利用DAzLE,我们从经50 mM氯化钾介导的膜去极化处理后的培养大鼠胚胎皮质神经元中分离出差异表达基因。我们鉴定出469个活动依赖型调控基因,其中174个是功能未知的基因。发现63个基因的调控依赖于一氧化氮(NO)。可识别的基因分为几个主要类别,包括信号转导途径、神经元发育、DNA复制、基因转录、蛋白质代谢、能量调节蛋白和抗凋亡蛋白。这些基因可能在活动依赖型神经元的存活和发育中起重要作用。此外,这些晚期反应基因提供了工具,以便开始研究活动依赖型神经元存活和发育中的下游事件。DAzLE的主要优点在于它提供了一个几乎完整且相对全面的差异筛选谱,有潜力成为其他研究领域强大且有用的工具。

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