Expression of a 12-kb promoter element derived from the zebrafish enolase-2 gene in the zebrafish visual system.
Expression of a 12-kb promoter element derived from the zebrafish enolase-2 gene in the zebrafish visual system.
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DOI：
10.1016/j.neulet.2008.10.101
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发表时间：
2009-01-16
影响因子：
2.5
通讯作者：
Burton EA
中科院分区：
文献类型：
--
作者：
Bai Q;Wei X;Burton EA
我们最近克隆了斑马鱼神经元烯醇酶-2基因，并发现一个12kb的enol2启动子元件足以在体内从受精后48小时到成年期在中枢神经系统神经元中广泛驱动转基因表达。本研究的目的是建立12kb eno2启动子元件在斑马鱼视觉系统中的表达模式。从受精后2天开始，在发育中的视网膜中检测到内源性eno2 mRNA，并通过12dpf定位于视网膜神经节细胞内、外核层。与内源性eno2相似，Tg(eno2:egfp)Pt404幼虫视网膜中GFP的表达在2dpf时首先明显，到12dpf时，GFP在视网膜神经节细胞层和光感受器层表达强烈，在内核层表达较弱。我们通过两种互补的策略鉴定了表达eno2启动子元件的细胞类型：(i)对Tg(eno2:egfp)Pt404斑马鱼进行双标记免疫荧光分析，（ii）在12kb eno2启动子和杆状或锥体特异性启动子下表达红色荧光蛋白的双转基因斑马鱼。12kb的eno2启动子在视网膜神经节细胞、无毛细胞（包括一个共表达酪氨酸羟化酶的亚群）和杆状光感受器中表达。这些数据表明，应该在使用该启动子产生的疾病的转基因模型中寻找视力异常。由于荧光报告基因在神经元亚群中的特异性表达，Tg(eno2:egfp)Pt404和Tg（eno2:RFP）斑马鱼可能对研究视网膜层压、视觉系统中的神经元分化和突触形成有用。
We recently cloned the zebrafish neuronal enolase-2 gene and showed that a 12kb eno2 promoter element was sufficient to drive transgene expression widely in CNS neurons in vivo from 48 hours post-fertilization through adulthood. The aim of the present study was to establish the expression pattern of the 12kb eno2 promoter element in the zebrafish visual system. Endogenous eno2 mRNA was detected in the developing retina from 2 days post-fertilization (dpf), and by 12dpf was localized to the retinal ganglion cell, inner and outer nuclear layers. Similar to endogenous eno2, GFP expression in the retina of Tg(eno2:egfp)Pt404 larvae was first evident at 2dpf, and by 12dpf intense GFP expression was seen in the retinal ganglion cell and photoreceptor layers, with weaker expression in the inner nuclear layer. We identified cell types expressing the eno2 promoter element by using two complementary strategies: (i) double label immunofluorescence analysis of Tg(eno2:egfp)Pt404 zebrafish, and (ii) generation of double transgenic zebrafish expressing red fluorescent protein under transcriptional control of the 12kb eno2 promoter and GFP under a rod- or cone-specific promoter. The 12kb eno2 promoter was expressed in retinal ganglion cells, amacrine cells, including a subset that co-expressed tyrosine hydroxylase, and rod photoreceptors. These data suggest that abnormalities of vision should be sought in transgenic models of diseases generated using this promoter. Owing to the specific expression of fluorescent reporters in neuronal subpopulations, Tg(eno2:egfp)Pt404 and Tg(eno2:RFP) zebrafish may be useful for studies of retinal lamination, neuronal differentiation and synapse formation in the visual system.
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影响因子：
16.2
作者：
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通讯作者：
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通讯作者：
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