Harvie P D, Filippova M, Bryant P J
Developmental Biology Center, University of California, Irvine, California 92697-2275, USA.
Genetics. 1998 May;149(1):217-31. doi: 10.1093/genetics/149.1.217.
We have used an enhancer-trap approach to begin characterizing the function of the Drosophila endocrine system during larval development. Five hundred and ten different lethal PZ element insertions were screened to identify those in which a reporter gene within the P element showed strong expression in part or all of the ring gland, the major site of production and release of developmental hormones, and which had a mutant phenotype consistent with an endocrine defect. Nine strong candidate genes were identified in this screen, and eight of these are expressed in the lateral cells of the ring gland that produce ecdysteroid molting hormone (EC). We have confirmed that the genes detected by these enhancer traps are expressed in patterns similar to those detected by the reporter gene. Two of the genes encode proteins, protein kinase A and calmodulin, that have previously been implicated in the signaling pathway leading to EC synthesis and release in other insects. A third gene product, the translational elongation factor EF-1alpha F1, could play a role in the translational regulation of EC production. The screen also identified the genes couch potato and tramtrack, previously known from their roles in peripheral nervous system development, as being expressed in the ring gland. One enhancer trap revealed expression of the gene encoding the C subunit of vacuolar ATPase (V-ATPase) in the medial cells of the ring gland, which produce the juvenile hormone that controls progression through developmental stages. This could reveal a function of V-ATPase in the response of this part of the ring gland to adenotropic neuropeptides. However, the gene identified by this enhancer trap is ubiquitously expressed, suggesting that the enhancer trap is detecting only a subset of its control elements. The results show that the enhancer trap approach can be a productive way of exploring tissue-specific genetic functions in Drosophila.
我们采用了增强子捕获方法来开始研究果蝇幼虫发育过程中内分泌系统的功能。我们筛选了510种不同的致死性PZ元件插入,以鉴定那些P元件中的报告基因在部分或全部环腺(发育激素产生和释放的主要部位)中表现出强表达,且具有与内分泌缺陷一致的突变表型的插入。在这个筛选中鉴定出了9个强候选基因,其中8个在产生蜕皮甾体蜕皮激素(EC)的环腺外侧细胞中表达。我们已经证实,这些增强子捕获检测到的基因的表达模式与报告基因检测到的相似。其中两个基因编码的蛋白质,蛋白激酶A和钙调蛋白,先前已被证明参与了其他昆虫中导致EC合成和释放的信号通路。第三个基因产物,翻译延伸因子EF-1α F1,可能在EC产生的翻译调控中发挥作用。该筛选还鉴定出了之前已知在周围神经系统发育中起作用的基因“couch potato”和“tramtrack”在环腺中表达。一个增强子捕获显示,编码液泡ATP酶(V-ATP酶)C亚基的基因在环腺中间细胞中表达,这些细胞产生控制发育阶段进程的保幼激素。这可能揭示了V-ATP酶在环腺这一部分对促性腺神经肽的反应中的功能。然而,这个增强子捕获鉴定出的基因是普遍表达的,这表明增强子捕获仅检测到了其控制元件的一个子集。结果表明,增强子捕获方法可以成为探索果蝇组织特异性遗传功能的有效途径。