Structural and functional characterization of mouse glutamate decarboxylase 67 gene promoter
Structural and functional characterization of mouse glutamate decarboxylase 67 gene promoter
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DOI：
10.1016/s0167-4781(03)00138-6
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发表时间：
2003-08-25
期刊：
影响因子：
--
通讯作者：
Yanagawa, Y
中科院分区：
文献类型：
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作者：
Kobayashi, T;Ebihara, S;Yanagawa, Y
小鼠谷氨酸脱羧酶67 （mGAD67）基因的神经元表达仅发生在合成和释放GABA的神经元（GABA能神经元）中。该基因也在胰岛细胞和睾丸精母细胞中表达。为了阐明mGAD67基因表达调控的分子机制，我们分离并表征了该基因的5'侧区。对该基因10.2 kb的DNA片段进行序列分析，其中包含一个启动子区域（8.4 kb）和非编码外显子0A和OB，结果显示存在许多潜在的神经元特异性顺式调控元件。通过瞬时转染培养细胞对外显子0A和OB的5'侧区域进行功能分析，发现外显子0A附近的-98 ~ -52区域对外显子0A和0B的转录活性都很重要。此外，我们用转基因小鼠检测了mGAD67基因10.2 kb DNA片段与细菌lacZ报告基因融合后的表达模式。在含有丰富gaba能神经元的特定脑区（如基底神经节）、胰岛细胞、睾丸精母细胞和精原细胞的神经元中观察到转基因表达。这些结果表明，mGAD67基因的10.2 kb DNA片段包含其在gaba能神经元、胰岛细胞和精母细胞中的靶向表达所必需的调控元件。(C) 2003 Elsevier B.V.版权所有
Neuronal expression of the mouse glutamate decarboxylase 67 (mGAD67) gene occurs exclusively in neurons that synthesize and release GABA (GABAergic neurons). This gene is also expressed in pancreatic islet cells and testicular spermatocytes. In order to elucidate the molecular mechanisms underlying the regulation of mGAD67 gene expression, we isolated and characterized the 5'-flanking region of this gene. Sequence analysis of a 10.2-kb DNA fragment of this gene containing a promoter region (8.4 kb) and noncoding exons 0A and OB revealed the presence of numerous potential neuron-specific cis-regulatory elements. Functional analysis of the 5'-flanking region of exons 0A and OB by transient transfection into cultured cells revealed that the region -98 to -52 close to exon 0A is important for the transcriptional activity of both exons 0A and 0B. In addition, we used transgenic mice to examine the expression pattern conferred by the 10.2 kb DNA fragment of the mGAD67 gene fused to the bacterial lacZ reporter gene. Transgene expression was observed in neurons of particular brain regions containing abundant GABAergic neurons such as the basal ganglia, in pancreatic islet cells and in testicular spermatocytes and spermatogonia. These results suggest that the 10.2 kb DNA fragment of the mGAD67 gene contains regulatory elements essential for its targeted expression in GABAergic neurons, islet cells and spermatocytes. (C) 2003 Elsevier B.V. All rights reserved.