RNA-sequencing of Chronic Phase Shift Protocol on Murine Pancreas

The objective of this study was to understand the long-term effects of a chronic phase shift protocol on the pancreatic transcriptome. A total of 144 wild-type C57Bl/6J 4-6 week old mice were housed under normal circadian conditions or disrupted using a chronic phase shift protocol whereby the light-dark cycle is advanced 8 hour every 2-3 days for 4 weeks. 12 hours prior to sacrifice mice were normalized under normal circadian conditions. Starting at ZT 0, mice were serially sacrificed at 4-hour intervals for 48 hours and the pancreas extracted. Pancreatic RNA was isolated and subjected to RNA-seq. A total of 144 samples were collected. Males (n = 3) and females (n=3) from each time point were pooled prior to sequencing. A total of 6 mice were sacrificed for each condition (normal circadian and circadian disrupted) at each time point.本研究旨在探究慢性相位偏移方案对胰腺转录组的长期影响。共计144只4~6周龄的野生型C57Bl/6J小鼠，被分为两组，分别饲养于正常昼夜节律环境，或通过慢性相位偏移方案实施节律干扰：该方案为每2~3天将光暗周期提前8小时，持续4周。在处死小鼠前12小时，将所有小鼠置于正常昼夜节律环境中进行节律归一化处理。从授时因子时间（Zeitgeber Time，ZT）0时刻起，每隔4小时处死一批小鼠，连续采样48小时，并采集胰腺组织。分离胰腺组织的RNA，并进行RNA测序（RNA-seq）。共计收集144份样本。每个时间点的雄性小鼠（n=3）与雌性小鼠（n=3）混合后进行测序。在每个时间点，两种实验条件（正常昼夜节律组、节律干扰组）各处死6只小鼠。