Zeng Chaoyang, Du Yanping, Alberico Thomas, Seeberger Jeanne, Sun Xiaoping, Zou Sige
University of Wisconsin at Milwaukee, Milwaukee, WI, USA.
Fly (Austin). 2011 Jul-Sep;5(3):174-80. doi: 10.4161/fly.5.3.15572. Epub 2011 Jul 1.
Drosophila melanogaster is ideal for studying lifespan modulated by dietary restriction (DR) and oxidative stress, and also for screening prolongevity compounds. It is critical to measure food intake in the aforementioned studies. Current methods, however, overlook the amount of the food excreted out of the flies as feces or deposited in eggs. Here we describe a feeding method using a radioactive tracer to measure gender-specific food intake, retention and excretion in response to DR and oxidative stress to account for all the ingested food. Flies were fed a full, restricted or paraquat-containing diet. The radioactivity values of the food in fly bodies, feces and eggs were measured separately after a 24-hr feeding. Food intake was calculated as the sum of these measurements. We found that most of the tracer in the ingested food was retained in the fly bodies and < 8% of the tracer was excreted out of the flies as feces and eggs in the case of females during a 24-hr feeding. Under a DR condition, flies increased food intake in volume to compensate for the reduction of calorie content in the diet and also slightly increased excretion. Under an oxidative stress condition, flies reduced both food intake and excretion. Under all the tested dietary conditions, males ingested and excreted 3-5 fold less food than females. This study describes an accurate method to measure food intake and provides a basis to further investigate prandial response to DR and prolongevity interventions in invertebrates.
黑腹果蝇是研究饮食限制(DR)和氧化应激调节寿命的理想模型,也是筛选延长寿命化合物的理想模型。在上述研究中,测量食物摄入量至关重要。然而,目前的方法忽略了果蝇以粪便形式排出或沉积在卵中的食物量。在这里,我们描述了一种使用放射性示踪剂的喂食方法,以测量在DR和氧化应激条件下,果蝇的性别特异性食物摄入量、保留量和排泄量,从而计算出所有摄入食物的总量。果蝇被喂食全量、限制量或含百草枯的食物。喂食24小时后,分别测量果蝇体内、粪便和卵中食物的放射性值。食物摄入量通过这些测量值的总和来计算。我们发现,在24小时的喂食过程中,摄入食物中的大部分示踪剂保留在果蝇体内,雌性果蝇以粪便和卵的形式排出的示踪剂不到8%。在DR条件下,果蝇增加食物摄入量以补偿饮食中卡路里含量的减少,同时排泄量也略有增加。在氧化应激条件下,果蝇的食物摄入量和排泄量均减少。在所有测试的饮食条件下,雄性果蝇的食物摄入量和排泄量比雌性果蝇少3-5倍。本研究描述了一种测量食物摄入量的准确方法,并为进一步研究无脊椎动物对DR和延长寿命干预的进食反应提供了基础。