Jacobsen Thomas L, Cain Donna, Paul Litty, Justiniano Steven, Alli Anwar, Mullins Jeremi S, Wang Chun Ping, Butchar Jon P, Simcox Amanda
Department of Molecular Genetics, The Ohio State University, Columbus, Ohio 43210, USA.
Genetics. 2006 Dec;174(4):1973-82. doi: 10.1534/genetics.106.056788. Epub 2006 Oct 8.
Differential gene expression is the major mechanism underlying the development of specific body regions. Here we assessed the role of genes differentially expressed in the Drosophila wing imaginal disc, which gives rise to two distinct adult structures: the body wall and the wing. Reverse genetics was used to test the function of uncharacterized genes first identified in a microarray screen as having high levels of expression in the presumptive wing. Such genes could participate in elaborating the specific morphological characteristics of the wing. The activity of the genes was modulated using misexpression and RNAi-mediated silencing. Misexpression of eight of nine genes tested caused phenotypes. Of 12 genes tested, 10 showed effective silencing with RNAi transgenes, but only 3 of these had resulting phenotypes. The wing phenotypes resulting from RNAi suggest that CG8780 is involved in patterning the veins in the proximal region of the wing blade and that CG17278 and CG30069 are required for adhesion of wing surfaces. Venation and apposition of the wing surfaces are processes specific to wing development providing a correlation between the expression and function of these genes. The results show that a combination of expression profiling and tissue-specific gene silencing has the potential to identify new genes involved in wing development and hence to contribute to our understanding of this process. However, there are both technical and biological limitations to this approach, including the efficacy of RNAi and the role that gene redundancy may play in masking phenotypes.
差异基因表达是特定身体区域发育的主要机制。在这里,我们评估了在果蝇翅芽盘中差异表达的基因的作用,翅芽盘会发育成两种不同的成虫结构:体壁和翅膀。反向遗传学被用于测试在微阵列筛选中首次鉴定出的、在假定的翅膀中具有高表达水平的未表征基因的功能。这些基因可能参与塑造翅膀的特定形态特征。通过错误表达和RNAi介导的沉默来调节基因的活性。测试的9个基因中有8个的错误表达导致了表型。在测试的12个基因中,10个通过RNAi转基因显示出有效的沉默,但其中只有3个产生了表型。RNAi导致的翅膀表型表明,CG8780参与了翅脉在翅片近端区域的图案形成,而CG17278和CG30069是翅膀表面黏附所必需的。翅脉形成和翅膀表面的并合是翅膀发育特有的过程,这为这些基因的表达和功能之间提供了一种关联。结果表明,表达谱分析和组织特异性基因沉默相结合有可能鉴定出参与翅膀发育的新基因,从而有助于我们对这一过程的理解。然而,这种方法存在技术和生物学上的局限性,包括RNAi的效率以及基因冗余可能在掩盖表型中所起的作用。