Glutamate receptor RNA editing: a molecular analysis of GluR2, GluR5 and GluR6 in human brain tissues and in NT2 cells following in vitro neural differentiation
Glutamate receptor RNA editing: a molecular analysis of GluR2, GluR5 and GluR6 in human brain tissues and in NT2 cells following in vitro neural differentiation
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DOI：
10.1016/s0169-328x(03)00317-6
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发表时间：
2003-10-07
期刊：
影响因子：
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通讯作者：
Barlati, S
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文献类型：
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作者：
Barbon, A;Vallini, I;Barlati, S
一些谷氨酸受体的特性可以通过 RNA 编辑来改变。这种转录后机制涉及谷氨酸受体前体 mRNA 中特定腺苷的酶促脱氨作用，由特定的 RNA 腺苷脱氨酶 (ADAR) 执行。这一事件导致基因编码的氨基酸被另一种氨基酸取代，从而改变了离子通道的生理特性。在此，我们报告了人畸胎癌细胞系 (NT2) 体外神经分化过程中 AMPA GluR2、红藻氨酸 GluR5 和 GluR6 编辑水平的分析，以及 GluR 和 ADAR 基因表达水平的分析。使用基于序列分析的特定标准化测定来分析编辑水平。该测定可以在所有编辑位点上进行，具有高水平的灵敏度和重现性。尽管 GluR 基因表达在 NT2 神经分化过程中增加，但 ADAR 基因的表达即使在未分化的 NT2 细胞中也可以以相当的水平检测到，并在分化过程中保持相对稳定。此外，大多数谷氨酸受体编辑位点在 NT2 神经分化过程中增加了其编辑水平，这表明 ADAR mRNA 的水平与分析的 GluR 中检测到的可变编辑水平并不密切相关。在人脑组织中，不同区域的编辑水平似乎受到精细调节，表明可能形成具有不同功能特性的离子通道，从而产生复杂的组织特异性受体调节和兴奋性刺激调节。 (C) 2003 Elsevier B.V. 保留所有权利。
The properties of some glutamate receptors are modified by RNA editing. This post-transcriptional mechanism involves the enzymatic deamination of specific adenosines in the pre-mRNA of the glutamate receptors, performed by specific RNA adenosine deaminases (ADARs). This event gives rise to the substitution of a gene-encoded amino acid with a different one that modifies the physiological properties of the ion channel. Here we report an analysis of the editing levels of AMPA GluR2, and kainate GluR5 and GluR6 in a human teratocarcinoma cell line (NT2) during in vitro neural differentiation, in conjunction with an analysis of the expression levels of GluR and ADAR genes. The editing levels were analysed using a specific standardised assay based on sequence analysis. This assay can be performed on all editing sites with a high level of sensitivity and reproducibility. Whereas GluR gene expression increased during NT2 neural differentiation, the expression of ADAR genes may be detected at comparable levels even in undifferentiated NT2 cells, remaining relatively stable during the differentiation process. Furthermore, most of the glutamate receptor editing sites increased their editing levels during NT2 neural differentiation, suggesting that the level of ADAR mRNAs is not closely related to the variable editing levels detected in the GluRs analysed. In human brain tissues, the editing levels appeared finely modulated in the different areas, indicating the possible formation of ion channels with different functional properties, thus generating a complex tissue-specific regulation of receptors and modulation of excitatory stimuli. (C) 2003 Elsevier B.V. All rights reserved.