Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase
Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase
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DOI：
10.1002/neu.10185
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发表时间：
2003-03-01
期刊：
影响因子：
--
通讯作者：
Birman, S
中科院分区：
文献类型：
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作者：
Friggi-Grelin, F;Coulom, H;Birman, S
多巴胺(DA)是果蝇体内唯一的儿茶酚胺能神经递质。在果蝇和其他昆虫的幼虫和成虫的中枢神经系统(CNS)中已经发现了多巴胺能神经元，但目前还没有专门的遗传工具来研究它们在体内的发育、功能和退化。在果蝇和脊椎动物中，DA生物合成的限速步骤是由酪氨酸羟基酶(TH)催化的。果蝇TH基因(DTH)在所有的多巴胺能细胞中特异表达，相应的突变体pAY(Ple)是胚胎致死的。我们已经用改良的DTH转基因进行了馅饼拯救实验。我们的结果表明，位于DTH内含子中的部分冗余调控元件是该基因在中枢神经系统中正常表达所必需的。在这项研究的基础上，我们获得了一个GAL4驱动基因TH-GAL4，它包含了DTH5‘侧翼和下游编码区的调控序列。TH-GAL4被导入果蝇基因组后，在胚胎、幼虫中枢神经系统和成年脑的多巴胺能细胞中特异表达。作为该驱动器的第一次应用，我们观察到在体内抑制DA释放在成年果蝇中诱导了显著的过度兴奋行为。我们认为TH-GAL4将有助于研究DA在果蝇行为和疾病模型中的作用。(C)2003年威利期刊公司。
Dopamine (DA) is the only catecholaminergic neurotransmitter in the fruit fly Drosophila melanogaster. Dopaminergic neurons have been identified in the larval and adult central nervous system (CNS) in Drosophila and other insects, but no specific genetic tool was available to study their development, function, and degeneration in vivo. In Drosophila as in vertebrates, the rate-limiting step in DA biosynthesis is catalyzed by the enzyme tyrosine hydroxylase (TH). The Drosophila TH gene (DTH) is specifically expressed in all dopaminergic cells and the corresponding mutant, pale (ple), is embryonic lethal. We have performed pie rescue experiments with modified DTH transgenes. Our results indicate that partially redundant regulatory elements located in DTH introns are required for proper expression of this gene in the CNS. Based on this study, we generated a GAL4 driver transgene, TH-GAL4, containing regulatory sequences from the DTH 5' flanking and downstream coding regions. TH-GAL4 specifically expresses in dopaminergic cells in embryos, larval CNS, and adult brain when introduced into the Drosophila genome. As a first application of this driver, we observed that in vivo inhibition of DA release induces a striking hyperexcitability behavior in adult flies. We propose that TH-GAL4 will be useful for studies of the role of DA in behavior and disease models in Drosophila. (C) 2003 Wiley Periodicals, Inc.