Circadian rhythm in clock mutants. Circadian rhythm in clock mutants

One hundred ninety wildtype male C57BL/6J mice age 7-10 weeks were purchased from Jackson Laboratory and entrained to a 12:12 light:dark cycle for 2 weeks. Mice were placed in light-tight boxes on a 12:12 LD cycle for 4 weeks, then released into constant darkness. Starting 30 hours after entry into DD (CT18), tissues from 5 (skeletal muscle) or 10 (liver or SCN) wildtype mice were collected every 4 hours for 48 hours, for a total of 12 timepoints. At timepoints 34 through 58 hours in DD, tissues from age-matched male C57BL/6J Clock/Clock homozygous mutant mice that had been treated with the same light entrainment protocol as the wildtype were collected. Tissues were collected from 5 Clock/Clock mutants at each timepoint except for 34 and 46 hours after the onset of DD, when tissues from 10 Clock/Clock mice were collected and run as independent replicates. Mice were sacrificed by cervical dislocation, and the optic nerves were severed in complete darkness; brain dissection was performed using illumination from an infrared viewer (FJW Industries, Palatine, IL). SCNs were dissected out, pooled at a density of 5 per tube in 100 ul RNAlater (Ambion, Austin, TX), frozen on dry ice, and stored at –80 degrees C until use. This SuperSeries is composed of the SubSeries listed below. Overall design: This reference Series links data in the following related Series: GSE3746 Circadian skeletal muscle_wt and Clock mutants GSE3748 Circadian liver_wt and Clock mutants

19只7~10周龄的野生型雄性C57BL/6J小鼠购自杰克逊实验室（Jackson Laboratory），先于12小时光照:12小时黑暗的光周期下进行光驯化2周。随后将小鼠置于密闭光箱内，维持12:12光照-黑暗（LD）循环4周，之后转入持续黑暗（DD）环境。自进入持续黑暗环境后的30小时（即昼夜节律时间18，CT18）起，每隔4小时采集一次组织样本，持续48小时，共计12个时间点。其中骨骼肌样本采集自5只野生型小鼠，肝脏与视交叉上核（suprachiasmatic nucleus, SCN）样本则采集自10只野生型小鼠。在持续黑暗环境中第34至58小时的时间点，同步采集经与野生型小鼠相同光驯化方案处理的同月龄雄性C57BL/6J Clock/Clock纯合突变小鼠的组织样本。除持续黑暗起始后第34、46小时需采集10只突变小鼠的组织作为独立重复样本外，其余每个时间点均采集5只突变小鼠的组织。小鼠均通过颈椎脱臼法处死，且需在完全黑暗环境中切断视神经；脑组织解剖需借助红外成像仪（FJW Industries公司，帕拉廷，伊利诺伊州，美国）提供照明。视交叉上核被分离取出后，以每管5个的密度合并置于100 μl RNAlater（Ambion公司，奥斯汀，德克萨斯州，美国）中，经干冰速冻后于-80℃条件下保存直至实验使用。本超级数据集（SuperSeries）由以下子数据集（SubSeries）组成：实验设计概述：本参考数据集关联以下相关数据集：GSE3746 节律相关骨骼肌野生型与Clock突变小鼠数据集；GSE3748 节律相关肝脏野生型与Clock突变小鼠数据集。