Remodeling of gene expression during C. elegans hibernation. [Ribo-Seq]

In the wild, many animals respond to cold temperatures by entering hibernation. In the clinic, controlled cooling is used widely in transplantation and emergency medicine. Yet, the molecular mechanisms the cells use in response to cold remain largely unexplored. One aspect of cold adaptation is a global downregulation of protein synthesis. Here, we performed total RNA sequencing combined with ribosome profiling to assess specific mRNA levels and their translational status during Caenorhabditis elegans hibernation. Overall design: Total RNA sequencing and ribosome profiling analysis was performed on synchronzied wild-type animals that were grown at different temperatures. For the 20°C condition, animals were grown from the L1 stage to 1-day-old adults at 20°C before collection. For the 10°C condition, animals were grown from the L1 stage to 1-day-old adults at 20°C, after which they were grown for 2 hours at 10°C before collection. For the 4°C 1, 3, or 5 days conditions, animals were grown from the L1 stage to 1-day-old adults at 20°C, after which they were grown for 2 hours at 10°C followed by 1, 3, or 5 days at 4°C before collection.

在自然环境中，诸多动物会通过进入冬眠状态来应对低温。在临床场景中，控制性降温已广泛应用于器官移植与急诊医学领域。然而，细胞应对低温的分子机制在很大程度上仍未被探明。
低温适应的一个核心特征是蛋白质合成的全局性下调。本研究结合总RNA测序（total RNA sequencing）与核糖体谱分析（ribosome profiling），对秀丽隐杆线虫（Caenorhabditis elegans）冬眠过程中特定信使RNA（mRNA）的表达水平及其翻译状态进行了评估。
实验设计：本研究对在不同温度条件下培养的同步化野生型秀丽隐杆线虫开展了总RNA测序与核糖体谱分析。
20℃组：线虫从L1幼虫期开始在20℃下培养至1日龄成虫后收集样本。
10℃组：线虫从L1幼虫期开始在20℃下培养至1日龄成虫，随后在10℃下培养2小时后收集样本。
4℃1/3/5天组：线虫从L1幼虫期开始在20℃下培养至1日龄成虫，先在10℃下培养2小时，随后分别在4℃下培养1、3或5天后收集样本。