Identification of neuron‐specific promoters in Ciona intestinalis
Identification of neuron‐specific promoters in Ciona intestinalis
复制标题
海鞘神经元特异性启动子的鉴定
作者：
R. Yoshida;D. Sakurai;T. Horie;I. Kawakami;M. Tsuda;T. Kusakabe
我们分离了4个基因Ci-GαI1、Ci-ARR、Ci-vAChTP和Ci-vGAT的5‘侧翼区域，每个基因都在肠海鞘中枢神经系统的不同神经元中表达，并通过将绿色荧光蛋白融合构建体引入到海鞘胚胎中来检测它们的功能。Ci-Gαi1、Ci-Arr和Ci-vAChTP 5‘侧翼区驱动的报告基因概括了内源性基因的表达模式，而Ci-vGAT5’侧翼区驱动的报告基因在表达内源性基因的特定神经元亚群中表达。缺失分析表明，Ci-Gαi1启动子由多个调控模块组成，控制着不同类型细胞的表达。GFP荧光使海鞘幼虫不同组神经元的胞体和轴突得以可视化。这些启动子可以成为研究神经发育的分子机制以及海鞘中神经网络和功能的有力工具。创世纪39：130-140,2004。©2004 Wiley-Liss Inc.
We isolated 5′ flanking regions of four genes, Ci‐Gαi1, Ci‐arr, Ci‐vAChTP,, and Ci‐vGAT, each of which is expressed in distinct sets of neurons in the central nervous system of the ascidian Ciona intestinalis, and we examined their function by introducing green fluorescent protein (GFP)‐fusion constructs into Ciona embryos. The reporter gene driven by the 5′ flanking region of Ci‐Gαi1, Ci‐arr, and Ci‐vAChTP recapitulated the endogenous gene expression patterns, while that of Ci‐vGAT can drive GFP expression in particular subsets of neurons expressing the endogenous gene. Deletion analysis revealed that the Ci‐Gαi1 promoter consists of multiple regulatory modules controlling the expression in different types of cells. The GFP fluorescence enabled visualization of cell bodies and axons of different sets of neurons in ascidian larvae. These promoters can be a powerful tool for studying molecular mechanisms of neuronal development as well as neuron networks and functions in ascidians. genesis 39:130–140, 2004. © 2004 Wiley‐Liss, Inc.