Abstract

Following prolonged swimming, Caenorhabditis elegans cycle between active swimming bouts and inactive quiescent bouts. Swimming is exercise for C. elegans and here we suggest that inactive bouts are a recovery state akin to fatigue. It is known that cGMP-dependent kinase (PKG) activity plays a conserved role in sleep, rest, and arousal. Using C. elegans EGL-4 PKG, we first validate a novel learning-based computer vision approach to automatically analyze C. elegans locomotory behavior and an edge detection program that is able to distinguish between activity and inactivity during swimming for long periods of time. We find that C. elegans EGL-4 PKG function impacts timing of exercise-induced quiescent (EIQ) bout onset, fractional quiescence, bout number, and bout duration, suggesting that previously described pathways are engaged during EIQ bouts. However, EIQ bouts are likely not sleep as animals are feeding during the majority of EIQ bouts. We find that genetic perturbation of neurons required for other C. elegans sleep states also does not alter EIQ dynamics. Additionally, we find that EIQ onset is sensitive to age and DAF-16 FOXO function. In summary, we have validated behavioral analysis software that enables a quantitative and detailed assessment of swimming behavior, including EIQ. We found novel EIQ defects in aged animals and animals with mutations in a gene involved in stress tolerance. We anticipate that further use of this software will facilitate the analysis of genes and pathways critical for fatigue and other C. elegans behaviors.

摘要

长时间游泳后，秀丽隐杆线虫在活跃的游泳回合和不活跃的静止回合之间循环。游泳是秀丽隐杆线虫的运动，在这里我们建议不活动的较量是一种类似于疲劳的恢复状态。众所周知，cGMP 依赖性激酶 (PKG) 活性在睡眠、休息和唤醒中起着保守的作用。使用秀丽隐杆线虫EGL-4 PKG，我们首先验证了一种新的基于学习的计算机视觉方法来自动分析秀丽隐杆线虫的运动行为，以及一种能够区分长时间游泳期间活动和不活动的边缘检测程序。我们发现C. elegansEGL-4 PKG 功能影响运动诱发静止 (EIQ) 发作的时间、静止分数、发作次数和发作持续时间，表明先前描述的通路在 EIQ 发作期间参与。然而，EIQ 较量可能不会睡眠，因为在大多数 EIQ 较量期间动物都在进食。我们发现其他线虫所需的神经元遗传扰动睡眠状态也不会改变 EIQ 动态。此外，我们发现 EIQ 开始对年龄和 DAF-16 FOXO 功能敏感。总之，我们已经验证了行为分析软件，可以对游泳行为（包括 EIQ）进行定量和详细的评估。我们在老年动物和应激耐受基因突变的动物中发现了新的 EIQ 缺陷。我们预计进一步使用该软件将有助于分析对疲劳和其他线虫行为至关重要的基因和通路。