Kaneko Gen, Yoshinaga Tatsuki, Gribble Kristin E, Welch David M, Ushio Hideki
Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo; School of Arts and Sciences, University of Houston-Victoria;
School of Marine Biosciences, Department of Marine Biosciences, Kitasato University.
J Vis Exp. 2016 Jul 22(113):54126. doi: 10.3791/54126.
Rotifers are microscopic cosmopolitan zooplankton used as models in ecotoxicological and aging studies due to their several advantages such as short lifespan, ease of culture, and parthenogenesis that enables clonal culture. However, caution is required when measuring their survival time as it is affected by maternal age and maternal feeding conditions. Here we provide a protocol for powerful and reproducible measurement of the survival time in Brachionus rotifers following a careful synchronization of culture conditions over several generations. Empirically, poor synchronization results in early mortality and a gradual decrease in survival rate, thus resulting in weak statistical power. Indeed, under such conditions, calorie restriction (CR) failed to significantly extend the lifespan of B. plicatilis although CR-induced longevity has been demonstrated with well-synchronized rotifer samples in past and present studies. This protocol is probably useful for other invertebrate models, including the fruitfly Drosophila melanogaster and the nematode Caenorhabditis elegans, because maternal age effects have also been reported in these species.
轮虫是一种微小的世界性浮游动物,由于其寿命短、易于培养以及孤雌生殖能够进行克隆培养等多种优势,被用作生态毒理学和衰老研究的模型。然而,在测量它们的存活时间时需要谨慎,因为这会受到母体年龄和母体喂养条件的影响。在这里,我们提供了一个方案,用于在几代培养条件仔细同步后,对臂尾轮虫的存活时间进行有效且可重复的测量。根据经验,同步不佳会导致早期死亡和存活率逐渐下降,从而导致统计效力较弱。事实上,在这种情况下,尽管过去和现在的研究已经用同步良好的轮虫样本证明了热量限制(CR)可延长寿命,但热量限制(CR)未能显著延长褶皱臂尾轮虫的寿命。该方案可能对其他无脊椎动物模型有用,包括果蝇黑腹果蝇和线虫秀丽隐杆线虫,因为在这些物种中也报道了母体年龄效应。