Hwang Hye Jin, Cox Rachel T
Department of Biochemistry and Molecular Biology, Uniformed Services University, Bethesda, MD 20814.
Henry M. Jackson Foundation, Rockville, MD.
bioRxiv. 2024 Aug 19:2024.08.19.608713. doi: 10.1101/2024.08.19.608713.
oogenesis has long been an important model for understanding myriad cellular processes controlling development, RNA biology, and patterning. Flies are easily fed drugs to disrupt various molecular pathways. However, this is often done under poor nutrient conditions that adversely affect oogenesis, thus making analysis challenging. Cycloheximide is a widely used compound that binds to and stalls the ribosome, therefore reducing protein synthesis. Since egg production is a highly nutrient-dependent process, we developed a method to feed female a rich diet of yeast paste supplemented with cycloheximide to better determine the effect of cycloheximide treatment on oogenesis. We find flies readily consume cycloheximide-supplemented yeast paste. Males and females have reduced lifespans when maintained on cycloheximide, with males exhibiting a dose-dependent decrease. While females did not exhibit decreased egg-laying, their ovaries were smaller and the number of progeny reduced, indicating substandard egg quality. Finally, females fed cycloheximide have disrupted oogenesis, with smaller ovaries, missing ovariole stages, and an increase in apoptotic follicles. Together, these data support that reduced protein synthesis adversely affects oogenesis with a rich diet that provides optimal nutrient conditions. In addition, this method could be used more broadly to test the effect of other drugs on oogenesis without the confounding effects caused by poor nutrition.
卵子发生长期以来一直是理解控制发育、RNA生物学和模式形成的众多细胞过程的重要模型。果蝇很容易被喂食药物以破坏各种分子途径。然而,这通常是在对卵子发生产生不利影响的不良营养条件下进行的,因此使得分析具有挑战性。放线菌酮是一种广泛使用的化合物,它与核糖体结合并使其停滞,从而减少蛋白质合成。由于产卵是一个高度依赖营养的过程,我们开发了一种方法,给雌性果蝇喂食富含酵母糊并添加放线菌酮的食物,以更好地确定放线菌酮处理对卵子发生的影响。我们发现果蝇很容易食用添加了放线菌酮的酵母糊。当果蝇在放线菌酮环境中饲养时,雄性和雌性的寿命都会缩短,雄性表现出剂量依赖性的下降。虽然雌性果蝇的产卵量没有减少,但它们的卵巢较小,后代数量减少,表明卵子质量不合格。最后,喂食放线菌酮的雌性果蝇卵子发生受到破坏,卵巢较小,卵巢管阶段缺失,凋亡卵泡增加。总之,这些数据支持在提供最佳营养条件的丰富饮食下,蛋白质合成减少会对卵子发生产生不利影响。此外,这种方法可以更广泛地用于测试其他药物对卵子发生的影响,而不会受到营养不良造成的混杂影响。