Sirtuins may mediate temperature-induced metabolic remodeling in threespine stickleback

The molecular pathway governing metabolic remodeling in response to temperature in fishes is unknown. We tested the hypothesis that sirtuins, a family of NAD+ dependent lysine deacetylases, drives an increase in aerobic metabolism in response to cold acclimation in stickleback. Stickleback were acclimated to 5, 12 and 20°C and sirtuin activity measured in liver, oxidative and glycolytic muscles using a Promega SIRT-Glo Assay kit. Protein levels of SIRT1 and SIRT3 were quantified using custom antibodies and capillary western blotting. In a second set of experiments, sirtuin activity was inhibited by feeding with EX-527 or activated by feeding with the NAD+ pre-cursor, nicotinamide mononucleotide (NMN). Standard metabolic rate (SMR) was measured using intermittent flow respirometry. Mitochondrial respiration rates were measured in permeabilized liver tissue using a substrate-uncoupler-inhibitor titration protocol . Sirtuin activity significantly increased in response to cold acclimation i..., , # Sirtuins may mediate temperature-induced metabolic remodeling in threespine stickleback Dataset DOI: [10.5061/dryad.n8pk0p38d](https://doi.org/10.5061/dryad.n8pk0p38d) ## Description of the data and file structure Stickleback (*Gasterosteus aculeatus*) were acclimated to 5, 12, and 20°C for 13 weeks. Sirtuin activity was then measured in liver, oxidative, and glycolytic muscles using a Promega SIRT-Glo assay kit, and protein levels of SIRT1 and SIRT3 were quantified using custom antibodies and capillary western blotting. In a second set of experiments, sticklebacks were acclimated to 20°C for 12 weeks. A subset of fish were then moved into an environmental chamber where the temperature in the room was decreased by 3°C per day over 2 days and then by 4°C on day 3 to reach 10°C. After 14 days at 10ºC, bloodworms were supplemented with either the sirtuin inhibitor EX-527 or the sirtuin activator, NMN, mixed with 7% agar for 14 days. Standard metabolic rate was then measured using inte...,目前尚不明确鱼类响应温度变化时调控代谢重编程的分子通路。本研究验证了如下假说：依赖于烟酰胺腺嘌呤二核苷酸（NAD+）的赖氨酸去乙酰化酶家族——沉默信息调节因子（sirtuins），可驱动三刺棘鱼在冷驯化过程中提升有氧代谢水平。 研究人员将三刺棘鱼分别驯化至5℃、12℃与20℃环境，采用普洛麦格（Promega）SIRT-Glo检测试剂盒，对肝脏、氧化型肌肉与糖酵解型肌肉中的沉默信息调节因子活性进行测定；同时使用定制抗体与毛细管免疫印迹法，定量检测SIRT1与SIRT3的蛋白表达水平。 在第二组实验中，研究人员将三刺棘鱼驯化至20℃环境12周。随后将部分个体转移至环境舱，以每日3℃的速率降温，持续2天，第3天降温4℃，最终使环境温度达到10℃。在10℃环境适应14天后，用添加了7%琼脂的血虫分别投喂沉默信息调节因子抑制剂EX-527与激活剂烟酰胺单核苷酸（nicotinamide mononucleotide, NMN），持续14天。采用间歇式呼吸测热法测定标准代谢率（SMR, standard metabolic rate）；使用底物-解偶联剂-抑制剂滴定方案，对透化肝脏组织的线粒体呼吸速率进行检测。研究发现，沉默信息调节因子活性随冷驯化过程显著升高…… # 沉默信息调节因子可能介导三刺棘鱼的温度诱导代谢重编程 数据集DOI：[10.5061/dryad.n8pk0p38d](https://doi.org/10.5061/dryad.n8pk0p38d) ## 数据与文件结构说明 三刺棘鱼（*Gasterosteus aculeatus*）被驯化至5℃、12℃与20℃环境，为期13周。随后采用Promega SIRT-Glo检测试剂盒，对肝脏、氧化型肌肉与糖酵解型肌肉中的沉默信息调节因子活性进行测定，并通过定制抗体与毛细管免疫印迹法定量检测SIRT1与SIRT3的蛋白水平。 在第二组实验中，棘鱼先在20℃环境驯化12周。随后将部分个体转移至环境舱，以每日3℃的速率降温，持续2天，第3天降温4℃，最终使环境温度降至10℃。在10℃环境适应14天后，用混合了7%琼脂的血虫分别投喂沉默信息调节因子抑制剂EX-527与激活剂NMN，持续14天。随后采用间歇式呼吸测热法测定标准代谢率……