Cellular dissection of the spinal cord motor column by BAC transgenesis and gene trapping in zebrafish.
Cellular dissection of the spinal cord motor column by BAC transgenesis and gene trapping in zebrafish.
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DOI：
10.3389/fncir.2013.00100
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发表时间：
2013
影响因子：
3.5
通讯作者：
Kawakami K
中科院分区：
文献类型：
--
作者：
Asakawa K;Abe G;Kawakami K
细菌人工染色体（BAC）转基因和基因/增强子捕获是鉴定中枢神经系统（CNS）中遗传上确定的神经元群体的有效方法。在这里，我们将这些技术应用于斑马鱼（Danio rerio），以深入了解脊椎动物的轴向运动柱的细胞结构。首先，通过使用Mnx类同源结构域蛋白基因mnr 2b/mnx 2b的BAC，我们建立了在大部分马达柱中表达Gal 4FF转录激活因子的mnGFF 7转基因系。单细胞标记的Gal 4FF-表达细胞的mnGFF 7线，使详细调查的形态学特征的个别脊髓运动神经元，以及整体组织的运动柱在脊髓节段。其次，从一个大规模的基因陷阱屏幕，我们确定了转基因线标记离散亚群的脊髓运动神经元与Gal 4FF。这些线的分子表征导致ADAMTS 3基因的鉴定，该基因编码进化上保守的ADAMTS家族的肽酶，并在腹侧脊髓中动态表达。在此建立的转基因鱼，沿着被鉴定的基因，应该有助于理解脊椎动物脊髓运动柱的细胞和分子结构及其与肌肉的连接。
Bacterial artificial chromosome (BAC) transgenesis and gene/enhancer trapping are effective approaches for identification of genetically defined neuronal populations in the central nervous system (CNS). Here, we applied these techniques to zebrafish (Danio rerio) in order to obtain insights into the cellular architecture of the axial motor column in vertebrates. First, by using the BAC for the Mnx class homeodomain protein gene mnr2b/mnx2b, we established the mnGFF7 transgenic line expressing the Gal4FF transcriptional activator in a large part of the motor column. Single cell labeling of Gal4FF-expressing cells in the mnGFF7 line enabled a detailed investigation of the morphological characteristics of individual spinal motoneurons, as well as the overall organization of the motor column in a spinal segment. Secondly, from a large-scale gene trap screen, we identified transgenic lines that marked discrete subpopulations of spinal motoneurons with Gal4FF. Molecular characterization of these lines led to the identification of the ADAMTS3 gene, which encodes an evolutionarily conserved ADAMTS family of peptidases and is dynamically expressed in the ventral spinal cord. The transgenic fish established here, along with the identified gene, should facilitate an understanding of the cellular and molecular architecture of the spinal cord motor column and its connection to muscles in vertebrates.
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影响因子：
13.9
作者：
Fenno L;Yizhar O;Deisseroth K
通讯作者：
Deisseroth K
影响因子：
25
作者：
Gabriel, Jens Peter;Ausborn, Jessica;El Manira, Abdeljabbar
通讯作者：
El Manira, Abdeljabbar
影响因子：
2.7
作者：
Davison, Jon M.;Akitake, Courtney M.;Parsons, Michael J.
通讯作者：
Parsons, Michael J.
影响因子：
3.7
作者：
Faucherre A;López-Schier H
通讯作者：
López-Schier H
影响因子：
2.7
作者：
Davidson, AE;Balciunas, D;Ekker, SC
通讯作者：
Ekker, SC