Characterization of Drosophila fruitless-gal4 transgenes reveals expression in male-specific fruitless neurons and innervation of male reproductive structures
Characterization of Drosophila fruitless-gal4 transgenes reveals expression in male-specific fruitless neurons and innervation of male reproductive structures
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DOI：
10.1002/cne.20177
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发表时间：
2004-07-19
影响因子：
2.5
通讯作者：
Goodwin, SF
中科院分区：
文献类型：
--
作者：
Billeter, JC;Goodwin, SF
无果(Fru)基因作用于果蝇的中枢神经系统(CNS)，以建立雄性性行为。对该基因的遗传解剖表明，fru基因的启动子之一，P1，控制着男性特有的Fru(M)蛋白的空间和时间表达，这些蛋白对确定刻板印象的男性性行为至关重要。利用Gal4-表达系统，我们证明了FruP1启动子5‘调控区的一个16kb片段驱动了在幼虫和成虫中枢神经系统中表达Fru(M)的神经元中Gal4的表达。Fru(M)和Gal4反应报告基因的共定位表明，Fru(P1)-Gal4融合结构在先前表征的表达Fru(M)的神经元以及尚未被证明表达Fru(M)的中枢神经系统和周围神经系统的细胞中都能产生表达。Gal4阳性神经元分布于与FRU功能直接相关的腹部器官，特别是劳伦斯肌(MOL)和男性内部生殖器官。后者的神经支配来自已发现的Fru(M)-5-羟色胺能神经元。此外，我们还发现MOL神经肌肉接头是性二型的。最后，我们描述了Gal4在支配男性生殖结构的神经突起中的表达，这些结构被认为是fru功能的靶标。因此，分离控制Fru在中枢神经系统中表达的调控序列，为操纵Fru(M)表达的神经元和了解果蝇生殖行为的细胞学基础提供了有力的工具。(C)2004年Wiley-Liss公司
The fruitless (fru) gene acts in the central nervous system (CNS) of Drosophila melanogaster to establish male sexual behavior. Genetic dissection of the locus has shown that one of the fru gene's promoter, P1, controls the spatial and temporal expression of male-specific Fru(M) proteins critical to determining stereotypical male sexual behavior. By using the Gal4-expression system, we show that a 16-kb fragment of the fru P1 promoter's 5' regulatory region drives the expression of Gal4 in a subset of Fru(M)-expressing neurons within both the pupal and adult CNS. Colocalization of Fru(M) and a Gal4-responsive reporter shows that the fru(P1)-gal4 fusion construct generates expression in both previously characterized Fru(M)-expressing neurons as well as within cells of both the CNS and the peripheral nervous system that have not been demonstrated as Fru(M)-expressing. Gal4-expressing neurons are shown to innervate abdominal organs directly relevant to fru function; specifically, the muscle of Lawrence (MOL) and the male internal reproductive organs. Innervations of the latter are shown to originate from identified Fru(M)-serotonergic neurons. Furthermore, we show that the MOL neuromuscular junction is sexually dimorphic. Finally, we describe Gal4 expression in neurites innervating male reproductive structures that are hypothesized to be targets of fru function. Isolation of the regulatory sequences controlling the expression of fru in the CNS, therefore, provides a potent tool for the manipulation of Fru(M)-expressing neurons and for understanding the cellular basis of Drosophila reproductive behavior. (C) 2004 Wiley-Liss, Inc.