DNA microarray analysis and functional profile of pituitary transcriptome under core-clock protein BMAL1 control

To find BMAL1-regulated genes in mice pituitary gland we performed a differential microarray from wild-type vs Bmal1-/- knock-out mice Wild type and clock-deficient Bmal1-/- mice in the C57BL/6J were used at 8 to 12 weeks of age. All animals were housed in a 12-h light/12-h dark cycle (light/dark, 12:12). Mice were sacrificed by decapitation at ZT12 (light off), and three pools of three pituitaries per genotype were used to isolate total RNA. Pituitary glands taken from three mice from the same genotype were removed and frozen in liquid nitrogen until total RNA isolation.为筛选小鼠垂体中受BMAL1调控的基因，本研究针对野生型与节律缺陷型Bmal1基因敲除（Bmal1-/-）小鼠开展了差异微阵列（differential microarray）实验。实验所用小鼠为C57BL/6J背景个体，鼠龄为8至12周。所有实验动物均饲养于12小时光照/12小时黑暗的循环环境（光照时长:黑暗时长=12:12）中。于节律时间点ZT12（灯光熄灭时刻）通过断头法处死小鼠，每个基因型设置3份垂体混合样本，每份样本由3只同基因型小鼠的垂体混合制备，用于总RNA提取。将同基因型3只小鼠的垂体取出后，即刻置于液氮中冻存，直至开展总RNA提取操作。