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首页> 外文期刊>The European Journal of Neuroscience >Analysis of the murine 5-HT receptor gene promoter in vitro and in vivo.
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Analysis of the murine 5-HT receptor gene promoter in vitro and in vivo.

机译：小鼠5-HT受体基因启动子的体外和体内分析。

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Abstract The expression level of the 5-HT(1A) receptor gene (htr1a) in the central nervous system (CNS) is implicated in the aetiology and treatment of anxiety disorders and depression. Previous studies of the murine htr1a have revealed that its proximal promoter is GC rich and TATA-less. Several functional transcription factor binding sites, including MAZ and SP1 recognition sequences, have been identified. To further analyse the promoter of this receptor gene, additional upstream sequence information extending to -5.5 kb of the murine htr1a was generated and promoter fragments extending to -20 kb were analysed for activity in cell culture and transgenic animals. Promoter fragments greater than 4.5 kb in length were active in 5-HT(1A) receptor mRNA positive cells and inactive in 5-HT(1A) receptor mRNA negative cells. Smaller fragments were not able to confer this specificity. In agreement, using additive transgenesis to drive LacZ expression in vivo, CNS specific reporter gene expression was found with these longer constructs. Transgene expression in the 4.5- and 20-kb mouse lines resembled the endogenous htr1a expression pattern, whereas the 5.5-kb mouse lines surprisingly revealed strongly reduced expression. None of the three constructs was prone to confer ectopic expression, however, variation of expression between the transgenic lines was observed. Using colocalization studies we analysed the degree of concurrence of transgenic and endogenous htr1a expression brought about by these three different constructs. The highest degrees of colocalization were observed in mice harbouring the 20-kb construct, suggesting a large promoter fragment is required to faithfully direct transgene expression in a 5-HT(1A) receptor like pattern.
机译：摘要5-HT（1A）受体基因（htr1a）在中枢神经系统（CNS）中的表达水平与焦虑症和抑郁症的病因和治疗有关。先前对鼠htr1a的研究表明，其近端启动子富含GC，且不含TATA。已经鉴定了几个功能性转录因子结合位点，包括MAZ和SP1识别序列。为了进一步分析该受体基因的启动子，产生了延伸至鼠htr1a -5.5 kb的其他上游序列信息，并分析了延伸至-20 kb的启动子片段在细胞培养和转基因动物中的活性。长度大于4.5 kb的启动子片段在5-HT（1A）受体mRNA阳性细胞中有活性，而在5-HT（1A）受体mRNA阴性细胞中无活性。较小的片段不能赋予这种特异性。一致地，使用加性转基因在体内驱动LacZ表达，发现了这些更长的构建体的CNS特异性报告基因表达。 4.5和20 kb小鼠品​​系中的转基因表达与内源性htr1a表达模式相似，而5.5 kb小鼠品​​系中的转基因表达出人意料地显着降低了表达。三种构建体均不易于产生异位表达，但是，观察到转基因品系之间表达的变化。使用共定位研究，我们分析了这三种不同构建体带来的转基因和内源性htr1a表达的并发程度。在具有20-kb构建体的小鼠中观察到最高的共定位程度，表明需要大的启动子片段来忠实地指导转基因在5-HT（1A）受体样模式中的表达。

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