Ray Mukulika, Lakhotia Subhash C
Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221 005, India.
J Genet. 2015 Sep;94(3):407-16. doi: 10.1007/s12041-015-0535-8.
The binary GAL4-UAS system of conditional gene expression is widely used by Drosophila geneticists to target expression of the desired transgene in tissue of interest. In many studies, a preferred target tissue is the Drosophila eye, for which the sev-GAL4 and GMR-GAL4 drivers are most widely used since they are believed to be expressed exclusively in the developing eye cells. However, several reports have noted lethality following expression of certain transgenes under these GAL4 drivers notwithstanding the fact that eye is not essential for survival of the fly. Therefore, to explore the possibility that these drivers may also be active in tissues other than eye, we examined the expression of UAS-GFP reporter driven by the sev-GAL4 or GMR-GAL4 drivers. We found that both these drivers are indeed expressed in additional tissues, including a common set of specific neuronal cells in larval and pupal ventral and cerebral ganglia. Neither sev nor glass gene has so far been reported to be expressed in these neuronal cells. Expression pattern of sev-GAL4 driver parallels that of the endogenous Sevenless protein. In addition to cells in which sev-GAL4 is expressed, the GMR-GAL4 is expressed in several other larval cell types also. Further, two different GMR-GAL4 lines also show some specific differences in their expression domains outside the eye discs. These findings emphasize the need for a careful confirmation of the expression domains of a GAL4 driver being used in a given study, rather than relying only on the empirically claimed expression domains.
二元GAL4-UAS条件性基因表达系统被果蝇遗传学家广泛用于在感兴趣的组织中靶向表达所需的转基因。在许多研究中,一个首选的靶组织是果蝇眼睛,sev-GAL4和GMR-GAL4驱动子在其中使用最为广泛,因为人们认为它们仅在发育中的眼细胞中表达。然而,有几份报告指出,在这些GAL4驱动子下表达某些转基因后会出现致死性,尽管眼睛对果蝇的生存并非必不可少。因此,为了探究这些驱动子是否也可能在眼睛以外的组织中活跃,我们检测了由sev-GAL4或GMR-GAL4驱动子驱动的UAS-GFP报告基因的表达。我们发现这两种驱动子确实在其他组织中表达,包括幼虫和蛹的腹侧及脑神经节中的一组共同的特定神经元细胞。到目前为止,尚未有sev或玻璃基因在这些神经元细胞中表达的报道。sev-GAL4驱动子的表达模式与内源性Sevenless蛋白的表达模式相似。除了sev-GAL4表达的细胞外,GMR-GAL4在其他几种幼虫细胞类型中也有表达。此外,两种不同的GMR-GAL4品系在眼盘以外的表达域也显示出一些特定差异。这些发现强调了在特定研究中使用GAL4驱动子时,需要仔细确认其表达域,而不是仅依赖于经验性声称的表达域。