Identification of transcriptional regulatory elements for Ntng1 and Ntng2 genes in mice.
Identification of transcriptional regulatory elements for Ntng1 and Ntng2 genes in mice.
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DOI：
10.1186/1756-6606-7-19
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发表时间：
2014-03-19
期刊：
影响因子：
3.6
通讯作者：
Itohara S
中科院分区：
文献类型：
--
作者：
Yaguchi K;Nishimura-Akiyoshi S;Kuroki S;Onodera T;Itohara S
高级大脑功能由数千个基因的精确时间和空间调节支持。然而，大脑中转录调控的机制仍不清楚。Ntng 1和Ntng 2基因分别编码轴突膜粘附蛋白netrin-G1和netrin-G2，是在脊椎动物中进化的旁系同源物，并以互补的方式在不同的神经元亚群中表达。这些基因的特征性表达模式提供了哺乳动物皮质层结构的部分基础。我们使用基因靶向技术，细菌人工染色体（BAC）辅助转基因技术，并在体内增强子测定，以检查体内转录机制，以了解这些基因的特征表达模式是如何获得的。在netrin-G1和netrin-G2功能蛋白存在或不存在的情况下的基因表达模式的分析允许我们排除反馈或前馈机制介导其特征表达模式的可能性。BAC缺失序列的发现表明，广泛分布的顺式调控元件的组合决定了这些基因的差异基因表达模式，并且主要的顺式调控元件位于Ntng 2的85-45 kb上游区域和Ntng 1的75-60 kb上游区域和内含子区域。在体内增强子测定使用2-kb的进化保守区域检测增强子活性的远端上游区域的两个基因。Ntng 1和Ntng 2的互补表达模式由广泛分布在这些位点的转录顺式调控元件决定。在这项研究中的顺式调控元件的特点，将促进新的遗传工具的发展，功能解剖神经回路，以更好地了解脊椎动物的大脑功能。
Higher brain function is supported by the precise temporal and spatial regulation of thousands of genes. The mechanisms that underlie transcriptional regulation in the brain, however, remain unclear. The Ntng1 and Ntng2 genes, encoding axonal membrane adhesion proteins netrin-G1 and netrin-G2, respectively, are paralogs that have evolved in vertebrates and are expressed in distinct neuronal subsets in a complementary manner. The characteristic expression patterns of these genes provide a part of the foundation of the cortical layer structure in mammals. We used gene-targeting techniques, bacterial artificial chromosome (BAC)-aided transgenesis techniques, and in vivo enhancer assays to examine transcriptional mechanisms in vivo to gain insight into how the characteristic expression patterns of these genes are acquired. Analysis of the gene expression patterns in the presence or absence of netrin-G1 and netrin-G2 functional proteins allowed us to exclude the possibility that a feedback or feedforward mechanism mediates their characteristic expression patterns. Findings from the BAC deletion series revealed that widely distributed combinations of cis-regulatory elements determine the differential gene expression patterns of these genes and that major cis-regulatory elements are located in the 85–45 kb upstream region of Ntng2 and in the 75–60 kb upstream region and intronic region of Ntng1. In vivo enhancer assays using 2-kb evolutionarily conserved regions detected enhancer activity in the distal upstream regions of both genes. The complementary expression patterns of Ntng1 and Ntng2 are determined by transcriptional cis-regulatory elements widely scattered in these loci. The cis-regulatory elements characterized in this study will facilitate the development of novel genetic tools for functionally dissecting neural circuits to better understand vertebrate brain function.
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影响因子：
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作者：
Kim, Seho;Burette, Alain;Kim, Eunjoon
通讯作者：
Kim, Eunjoon
影响因子：
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Bryndorf, Thue
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作者：
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