Department of Biochemistry and Molecular Biology, Uniformed Services University, Bethesda, MD 20814, USA.
Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD 20817, USA.
Biol Open. 2024 Nov 15;13(11). doi: 10.1242/bio.061697. Epub 2024 Nov 25.
Drosophila 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. As egg production is a highly nutrient-dependent process, we developed a method to feed female Drosophila a rich diet of yeast paste supplemented with cycloheximide to better determine the effect of cycloheximide treatment on oogenesis. We found that flies readily consumed cycloheximide-supplemented yeast paste. Males and females had reduced lifespans when maintained on cycloheximide, with males exhibiting a dose-dependent decrease. Although 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 had 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 Drosophila oogenesis without the confounding effects caused by poor nutrition.
果蝇卵子发生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。可以很容易地给果蝇喂食药物来破坏各种分子途径。然而，这通常是在不良的营养条件下进行的，这些条件会对卵子发生产生不利影响，从而使分析变得具有挑战性。环己酰亚胺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。由于卵子发生是一个高度依赖营养的过程，我们开发了一种方法，即用富含酵母糊的食物喂养雌性果蝇，并补充环己酰亚胺，以更好地确定环己酰亚胺处理对卵子发生的影响。我们发现，果蝇很容易消耗含有环己酰亚胺的酵母糊。用环己酰亚胺维持的雄性和雌性果蝇的寿命都缩短了，雄性果蝇的寿命呈剂量依赖性下降。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，表明卵子质量较差。最后，喂食环己酰亚胺的雌性果蝇卵子发生受到干扰，卵巢变小，卵巢小管阶段缺失，凋亡滤泡增加。总之，这些数据表明，在提供最佳营养条件的丰富饮食中，减少蛋白质合成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果蝇卵子发生的影响，而不会受到营养不良引起的混杂影响。
