Department of Biological Sciences, Marquette University, P.O. Box 1881, Milwaukee, WI 53201-1881, USA.
Insect Biochem Mol Biol. 2012 Sep;42(9):690-8. doi: 10.1016/j.ibmb.2012.06.002. Epub 2012 Jun 21.
In Drosophila melanogaster, mutations in the gene drop-dead (drd) result in early adult lethality, with flies dying within 2 weeks of eclosion. Additional phenotypes include neurodegeneration, tracheal defects, starvation, reduced body mass, and female sterility. The cause of early lethality and the function of the drd protein remain unknown. In the current study, the temporal profiles of drd expression required for adult survival and body mass regulation were investigated. Knockdown of drd expression by UAS-RNAi transgenes and rescue of drd expression on a drd mutant background by a UAS-drd transgene were controlled with the Heat Shock Protein 70 (Hsp70)-Gal4 driver. Flies were heat-shocked at different stages of their lifecycle, and the survival and body mass of the resulting adult flies were assayed. Surprisingly, the adult lethal phenotype did not depend upon drd expression in the adult. Rather, expression of drd during the second half of metamorphosis was both necessary and sufficient to prevent rapid adult mortality. In contrast, the attainment of normal adult body mass required a different temporal pattern of drd expression. In this case, manipulation of drd expression solely during larval development or metamorphosis had no effect on body mass, while knockdown or rescue of drd expression during all of pre-adult (embryonic, larval, and pupal) development did significantly alter body mass. Together, these results indicate that the adult-lethal gene drd is required only during development. Furthermore, the mutant phenotypes of body mass and lifespan are separable phenotypes arising from an absence of drd expression at different developmental stages.
在黑腹果蝇中,基因突变导致 drop-dead(drd)基因的过早成年致死,突变果蝇在羽化后 2 周内死亡。其他表型包括神经退行性变、气管缺陷、饥饿、体重减轻和雌性不育。导致早期致死和 drd 蛋白功能的原因仍不清楚。在本研究中,研究了 drd 表达的时间谱,以了解其对成年生存和体重调节的影响。通过 UAS-RNAi 转基因敲低 drd 表达,并用 UAS-drd 转基因拯救 drd 突变背景中的 drd 表达,受热休克蛋白 70(Hsp70)-Gal4 驱动控制。在生命周期的不同阶段对果蝇进行热休克处理,然后检测产生的成年果蝇的存活和体重。令人惊讶的是,成年致死表型并不依赖于成年期的 drd 表达。相反,在变态的后半期表达 drd 既是必需的,也是防止快速成年死亡率的充分条件。相比之下,获得正常的成年体重需要不同的 drd 表达时间模式。在这种情况下,仅在幼虫发育或变态期间操纵 drd 表达对体重没有影响,而在所有前成年期(胚胎、幼虫和蛹期)发育过程中敲低或拯救 drd 表达则显著改变了体重。总之,这些结果表明,仅在发育过程中需要 adult-lethal 基因 drd。此外,体重和寿命的突变表型是可分离的表型,它们是由于不同发育阶段缺乏 drd 表达引起的。