Pharmacogenomic research for mood disorder

鉴定了常规5-羟色胺转运体(5-HTT)的三个变异体，它们由不同的外显子1(外显子1a、外显子1b或外显子1c)和相同的外显子2至外显子5组成。在某些细胞系中，连接到PGL-3增强子载体的任何启动子区域的活性都显著高于空载体，而对外显子-1c的活性显著低于其他细胞系。在COS-7细胞中，二丁酰环腺苷(Dib-cAMP)处理降低了外显子1c的活性，而不改变其他外显子-1c的活性。人干扰素处理降低了外显子1a的活性，但不改变外显子1b的活性，增加了外显子1c的活性。在PC12细胞中，外显子1a和外显子1b的启动子区域的活性高于模拟载体。外显子-L c的启动子活性与模拟载体相同。DIB-cAMP处理增加了所有构建体的活性，而应用人INF-α不改变任何…在永生化的大鼠中缝神经元RN46a中，插入外显子1a、外显子1b和外显子1c的启动子区域后，活性分别提高了12倍、16倍和3倍。DIB-cAMP略微增加了它们的活性，而作为小鼠干扰素*不能改变它们。这三个启动子区域可能在5-HTT的转录过程中发挥作用，并可能为调节人类5-HTT的产生以及进一步研究抑郁症和其他5-羟色胺谱系障碍的发病机制提供一个模型。5-HTTLR变异体连接到PGL-3启动子1载体后，COS-7细胞和PC12细胞中的水解酶活性增加，但差异无统计学意义。然而，在RN46A细胞系中，5-HTTLPR将水解酶活性降低到80%到30%，起到了沉默而不是促进的作用。一些等位变异(15、19、20和22)表现出比RN46A中的其他变异体更强的沉默活性。我们还检查了等位基因频率、增强器/消音器活动和情绪障碍事件之间的关系。心境障碍组的等位基因频率与心境障碍组比较差异无统计学意义。小鼠5-羟色胺转运体(5-HTT)基因有3个变异体，分别由不同的外显子1(外显子1a、外显子1b或外显子1c)和相同的外显子4w0至外显子5组成。在某些细胞系中，连接到PGL-3增强子载体的任何启动子区域的活性都显著高于空载体，而对外显子-1c的活性显著低于其他细胞系。较少

Three variants of routine serotonin transporter (5-HTT) mRNA, which consist of a different exon-one (exon-la, exon-1b or exon-1c) and the same exon-two to exon-five, were identified. Any promoter region, ligated to PGL-3 enhancer vector, had activities significantly higher than the empty vector in some cell lines tested, where as the activity for the exon-1c was significantly lower than the others. In COS-7 cell lines, dibutyryl-cyclic AMP (Dib-cAMP) treatment decreased the activity for the exon-lc and did not change the others. Human interferon-*** (ENF-***) treatment decreased the activity for the exon-la, did not change the one for the exon-lb and increased the one for exon-lc. In PC12 cell lines the promoter regions for exon-la and exon-lb had higher activities than the mock vector. The promoter activity for exon-l c did not differ from that of mock vector. Dib-cAMP treatment increased the activities of all of the constructs whereas human INF-α application did not change any of the … More m. In immortalized rat serotoneigic raphe neurons, RN46A, the insertion of the promoter region for exonl a, exonlb and exonlc increased the activity twelve, sixteen or three times respectively. Dib-cAMP slightly increased their activity where as murine interferon *** did not change them. These three promoter regions may play a role in transcription of 5-HTT and could offer a model of the regulation of 5-HTT production in human and further the pathogenesis of depression and other serotonin spectrum disorders.Enhancer/silencer activity of each allelic variant of the human serotonin transporter linked polymorphic region (5HTTLPR) including newly found ones was measured in several cell lines including raphe-nucleus-derived RN46A. 5-HTTLR variants ligated in pGL-3 promote1 vector increased hiciferase activity in COS-7 cells and PC12 cells, where no significant differences among the variants were observed. In RN46A cell lines, however, 5-HTTLPRs decreased hiciferase activity to eighty to thirty percent, acting as silencers not as enhancers. Some allelic variants (15, 19, 20, and 22) showed even significantly stronger silencer activities than others in RN46A. We also examined relationship between allelic frequencies, the enhancer/silencer activities and incidents of mood disorder. The categorized genotypic or allelic frequencies was not significantly different between the mood disorder and the control. No significant difference was detected either when analyzed by silencer activities of each allelic variant.Three variants ofmurine serotonin transporter (5-HTT) mRNA, which consist of a different exon-one (exon-la, exon-lb or exon-lc) and the same exon4wo to exon-five, were identified. Any promoter region, ligated to pGL-3 enhancer vector, had activities significantly higher than the empty vector in some cell lines tested, where as the activity for the exon-lc was significantly lower than the others. Less