Identification of neural crest and glial enhancers at the mouse Sox10 locus through transgenesis in zebrafish.
Identification of neural crest and glial enhancers at the mouse Sox10 locus through transgenesis in zebrafish.
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DOI：
10.1371/journal.pgen.1000174
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发表时间：
2008-09-05
期刊：
影响因子：
4.5
通讯作者：
Pavan, William J.
中科院分区：
文献类型：
--
作者：
Antonellis, Anthony;Huynh, Jimmy L.;Lee-Lin, Shih-Queen;Vinton, Ryan M.;Renaud, Gabriel;Loftus, Stacie K.;Elliot, Gene;Wolfsberg, Tyra G.;Green, Eric D.;McCallion, Andrew S.;Pavan, William J.
Sox10是一个动态调节的转录因子基因，对神经嵴衍生和少突胶质细胞群体的发育至关重要。发育基因通常需要多个调控序列，这些调控序列整合了离散和重叠的功能以协调它们的表达。为了鉴定Sox10顺式调控元件，我们整合了多个模型系统，包括基于细胞的筛选和转座子介导的斑马鱼转基因，以仔细检查小鼠Sox10基因座的哺乳动物保守的非编码基因组片段。我们证明，11个Sox10基因组元素中的8个直接报告基因在转基因斑马鱼中的表达类似于在转基因小鼠中观察到的模式，尽管小鼠和斑马鱼之间没有可观察到的序列保守性。多个片段指导神经嵴衍生物和神经胶质细胞重叠群体中的表达，范围从泛Sox10和泛神经嵴调节控制到Sox10表达细胞亚群中的表达调节，包括发育中的黑素细胞和许旺细胞。几个序列表现出重叠的空间控制，但在不完全重叠的发育间隔直接表达。我们能够部分解释神经嵴的表达模式的存在下，头对头SoxE家族结合位点内的两个元素。此外，我们能够使用这种转录因子结合位点的签名，以确定相应的斑马鱼增强子在缺乏整体序列同源性。我们证明了斑马鱼转基因作为一个高保真的替代品在哺乳动物基因调控的解剖，特别是那些动态控制的发育表达的效用。神经嵴是一群胚胎迁移干细胞。它们在未来的脊髓顶部形成，并在发育的胚胎中迁移，形成许多不同的细胞，包括表皮色素细胞，头部的骨细胞和周围神经系统的神经细胞。在这项研究中，我们研究了负责表达SOX 10的基因组元件，SOX 10是一种动态表达的基因，对神经嵴发育至关重要。我们通过鉴定该基因周围的一小部分基因组DNA来分离SOX 10的候选调控元件，这些基因在进化过程中不会随着鸟类和哺乳动物基因组的变化而变化。我们测试了这些片段在斑马鱼中调节基因表达的能力，斑马鱼是DNA介导的表达研究和胚胎学的高效模型系统。我们发现，即使基因组序列与鱼类的SOX 10基因不相似，基因组片段也能够重现发育过程中SOX 10的动态表达。通过计算分析的序列，我们确定了一个转录因子结合位点的签名，确定了相应的斑马鱼SOX 10调控元件。这项研究描述了一个范例解剖调控的必需基因，显示复杂的表达模式在发展过程中。
Sox10 is a dynamically regulated transcription factor gene that is essential for the development of neural crest–derived and oligodendroglial populations. Developmental genes often require multiple regulatory sequences that integrate discrete and overlapping functions to coordinate their expression. To identify Sox10 cis-regulatory elements, we integrated multiple model systems, including cell-based screens and transposon-mediated transgensis in zebrafish, to scrutinize mammalian conserved, noncoding genomic segments at the mouse Sox10 locus. We demonstrate that eight of 11 Sox10 genomic elements direct reporter gene expression in transgenic zebrafish similar to patterns observed in transgenic mice, despite an absence of observable sequence conservation between mice and zebrafish. Multiple segments direct expression in overlapping populations of neural crest derivatives and glial cells, ranging from pan-Sox10 and pan-neural crest regulatory control to the modulation of expression in subpopulations of Sox10-expressing cells, including developing melanocytes and Schwann cells. Several sequences demonstrate overlapping spatial control, yet direct expression in incompletely overlapping developmental intervals. We were able to partially explain neural crest expression patterns by the presence of head to head SoxE family binding sites within two of the elements. Moreover, we were able to use this transcription factor binding site signature to identify the corresponding zebrafish enhancers in the absence of overall sequence homology. We demonstrate the utility of zebrafish transgenesis as a high-fidelity surrogate in the dissection of mammalian gene regulation, especially those with dynamically controlled developmental expression. The neural crest is a population of embryonic migratory stem cells. They form atop the future spinal cord and migrate throughout developing embryos and form many different cells, including the epidermal pigment cells, bone cells in the head, and nerve cells of the peripheral nervous system. In this study, we studied the genome elements responsible for expression of SOX10, a dynamically expressed gene that is essential for neural crest development. We isolated candidate regulatory elements for SOX10 by identifying the small percentage of genomic DNA around the gene that did not vary as avian and mammalian genomes changed though evolution. We tested these fragments for their ability to regulate gene expression in zebrafish, a model system that is highly efficient for DNA-mediated expression studies and embryology. We found that even though the genome sequences were not similar to the SOX10 gene in fish, the genomic fragments were able to recapitulate the dynamic expression of SOX10 during development. Through computational analysis of the sequences, we identified a transcription factor binding site signature that identified the corresponding zebrafish SOX10 regulatory elements. This study describes a paradigm for dissecting regulation of essential genes that display complex expression patterns during development.
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