Retinoic acid-mediated gene expression in transgenic reporter zebrafish
Retinoic acid-mediated gene expression in transgenic reporter zebrafish
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DOI：
10.1006/dbio.2000.9979
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发表时间：
2001-01-01
影响因子：
2.7
通讯作者：
Linney, E
中科院分区：
文献类型：
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作者：
Perz-Edwards, A;Hardison, NL;Linney, E
视黄酸介导的基因激活对于脊椎动物的正常发育是重要的。视黄酸的大小和性质使得很难确定这种信号分子在整个胚胎中的精确细胞位置。此外，视黄酸（RA）信号传导受受体、辅激活剂和拮抗蛋白的复杂组合调节。因此，为了整合这些信号，并确定在整个发育中的胚胎细胞可以响应于视黄酸转录的区域，我们使用了报告基因转基因方法。我们已经产生了几个稳定的转基因斑马鱼线，使用视黄酸响应元件驱动荧光蛋白的表达。在这些斑马鱼品系中，转基因表达定位于胚胎和幼虫的神经管、视网膜、脊索、体节、心脏、前肾管、鳃弓和颌肌的区域。通过将胚胎暴露于视黄酸，可以在神经管和视网膜的其他区域以及未成熟的脊索、孵化腺、包膜细胞层和鳍中诱导转基因表达。视黄酸合成酶抑制剂，柠檬醛和二乙氨基苯甲醛（DEAB）的治疗，在神经形成，大大降低了转基因表达。DEAB处理胚胎原肠胚phenocopies胚胎的影响，维生素A的剥夺或有针对性的破坏RA合酶的视黄醇脱氢酶-2在其他脊椎动物。这些数据表明，我们在斑马鱼中看到的报告基因表达依赖于保守的脊椎动物RA合成途径。(C)北京：科学出版社.
Retinoic acid-mediated gene activation is important for normal vertebrate development. The size and nature of retinoic acid make it difficult to identify the precise cellular location of this signaling molecule throughout an embryo. Additionally, retinoic acid (RA) signaling is regulated by a complex combination of receptors, coactivators, and antagonizing proteins. Thus, in order to integrate these signals and identify regions within a whole developing embryo where cells can respond transcriptionally to retinoic acid, we have used a reporter transgenic approach. We have generated several stable lines of transgenic zebrafish which use retinoic acid response elements to drive fluorescent protein expression. In these zebrafish lines, transgene expression is localized to regions of the neural tube, retina, notochord, somites, heart, pronephric ducts, branchial arches, and jaw muscles in embryos and larvae. Transgene expression can be induced in additional regions of the neural tube and retina as well as the immature notochord, hatching gland, enveloping cell layer, and fin by exposing embryos to retinoic acid. Treatment with retinoic acid synthase inhibitors, citral and diethylaminobenzaldehyde (DEAB), during neurulation, greatly reduces transgene expression. DEAB treatment of embryos at gastrulation phenocopies the embryonic effects of vitamin A deprivation or targeted disruption of the RA synthase retinaldehyde dehydrogenase-2 in other vertebrates. Together these data suggest that the reporter expression we see in zebrafish is dependent upon conserved vertebrate pathways of RA synthesis. (C) 2001 Academic Press.