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首页> 外文期刊>BMC Neuroscience >Both 5' and 3' flanks regulate Zebrafish brain-derived neurotrophic factor gene expression
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Both 5' and 3' flanks regulate Zebrafish brain-derived neurotrophic factor gene expression

机译：5'和3'侧翼均调节斑马鱼脑源性神经营养因子基因表达

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Background Precise control of developmental and cell-specific expression of the brain-derived neurotrophic factor (BDNF) gene is essential for normal neuronal development and the diverse functions of BDNF in the adult organism. We previously showed that the zebrafish BDNF gene has multiple promoters. The complexity of the promoter structure and the mechanisms that mediate developmental and cell-specific expression are still incompletely understood. Results Comparison of pufferfish and zebrafish BDNF gene sequences as well as 5' RACE revealed three additional 5' exons and associated promoters. RT-PCR with exon-specific primers showed differential developmental and organ-specific expression. Two exons were detected in the embryo before transcription starts. Of the adult organs examined, the heart expressed a single 5' exon whereas the brain, liver and eyes expressed four of the seven 5' exons. Three of the seven 5' exons were not detectable by RT-PCR. Injection of promoter/GFP constructs into embryos revealed distinct expression patterns. The 3' flank profoundly affected expression in a position-dependent manner and a highly conserved sequence (HCS1) present in 5' exon 1c in a dehancer-like manner. Conclusions The zebrafish BDNF gene is as complex in its promoter structure and patterns of differential promoter expression as is its murine counterpart. The expression of two of the promoters appears to be regulated in a temporally and/or spatially highly circumscribed fashion. The 3' flank has a position-dependent effect on expression, either by affecting transcription termination or post-transcriptional steps. HCS1, a highly conserved sequence in 5' exon 1c, restricts expression to primary sensory neurons. The tools are now available for detailed genetic and molecular analyses of zebrafish BDNF gene expression.
机译：背景技术精确控制脑源性神经营养因子（BDNF）基因的发育和细胞特异性表达对于正常的神经元发育和BDNF在成体生物体中的多种功能至关重要。我们以前表明斑马鱼BDNF基因具有多个启动子。启动子结构的复杂性和介导发育和细胞特异性表达的机制仍不完全了解。结果河豚和斑马鱼的BDNF基因序列以及5'RACE的比较显示了另外三个5'外显子和相关的启动子。使用外显子特异性引物的RT-PCR显示差异的发育和器官特异性表达。在转录开始之前，在胚胎中检测到两个外显子。在检查的成年器官中，心脏表达了一个5'外显子，而大脑，肝脏和眼睛表达了七个5'外显子中的四个。 RT-PCR无法检测到七个5'外显子中的三个。将启动子/ GFP构建体注射入胚胎揭示了不同的表达模式。 3'侧翼以位置依赖的方式深刻影响表达，而5'外显子1c中的高度保守序列（HCS1）则以类似dehancer的方式存在。结论斑马鱼BDNF基因的启动子结构和启动子差异表达模式与其鼠对应物一样复杂。两个启动子的表达似乎在时间和/或空间上高度外接的方式受到调节。 3'侧翼通过影响转录终止或转录后步骤，对表达具有位置依赖性。 HCS1，5'外显子1c中高度保守的序列，将表达限制在原代感觉神经元中。该工具现已可用于斑马鱼BDNF基因表达的详细遗传和分子分析。

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