Conserved Noncoding Sequences Regulate lhx5 Expression in the Zebrafish Forebrain.

Conserved Noncoding Sequences Regulate lhx5 Expression in the Zebrafish Forebrain.
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保守的非编码序列调节斑马鱼前脑中的 lhx5 表达。

DOI:
10.1371/journal.pone.0132525
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发表时间:
2015
期刊:
影响因子:
3.7
通讯作者:
Peng G
Peng G
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Sun L;Chen F;Peng G

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LIM 同源盒家族蛋白 Lhx5 在脊椎动物前脑发育中发挥重要作用。 lhx5基因在早期发育过程中表现出复杂的时空表达模式,但其转录调控机制尚不清楚。在这里,我们在斑马鱼中使用转基因来定义驱动前脑中 lhx5 表达的调控元件。通过比较基因组分析,我们鉴定了 5 个硬骨鱼物种中保守的 10 个非编码序列。接下来,我们通过 Tol2 转座子介导的转基因检查了这些保守非编码序列的增强子活性。我们发现位于附近的增强子在前脑区域产生了强大的报告 EGFP 表达。此外,我们还鉴定了位于 lhx5 编码区上游约 50 kb 处的增强子,该增强子负责下丘脑中报告基因的表达。我们还鉴定了位于丘脑前部(腹侧丘脑)表达所需的 lhx5 编码区上游约 40 kb 的增强子。我们的结果表明离散的增强子元件控制前脑不同区域的 lhx5 表达。
The LIM homeobox family protein Lhx5 plays important roles in forebrain development in the vertebrates. The lhx5 gene exhibits complex temporal and spatial expression patterns during early development but its transcriptional regulation mechanisms are not well understood. Here, we have used transgenesis in zebrafish in order to define regulatory elements that drive lhx5 expression in the forebrain. Through comparative genomic analysis we identified 10 non-coding sequences conserved in five teleost species. We next examined the enhancer activities of these conserved non-coding sequences with Tol2 transposon mediated transgenesis. We found a proximately located enhancer gave rise to robust reporter EGFP expression in the forebrain regions. In addition, we identified an enhancer located at approximately 50 kb upstream of lhx5 coding region that is responsible for reporter gene expression in the hypothalamus. We also identify an enhancer located approximately 40 kb upstream of the lhx5 coding region that is required for expression in the prethalamus (ventral thalamus). Together our results suggest discrete enhancer elements control lhx5 expression in different regions of the forebrain.
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