Hughes M, Deharo L, Pulivarthy S R, Gu J, Hayes K, Panda S, Hogenesch J B
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
Cold Spring Harb Symp Quant Biol. 2007;72:381-6. doi: 10.1101/sqb.2007.72.011.
In both the suprachiasmatic nucleus (SCN) and peripheral tissues, the circadian oscillator drives rhythmic transcription of downstream target genes. Recently, a number of studies have used DNA microarrays to systematically identify oscillating transcripts in plants, fruit flies, rats, and mice. These studies have identified several dozen to many hundred rhythmically expressed genes by sampling tissues every 4 hours for 1, 2, or more days. To extend this work, we have performed DNA microarray analysis on RNA derived from the mouse pituitary sampled every hour for 2 days. COSOPT and Fisher's G-test were used at a false-discovery rate of less than 5% to identify more than 250 genes in the pituitary that oscillate with a 24-hour period length. We found that increasing the frequency of sampling across the circadian day dramatically increased the statistical power of both COSOPT and Fisher's G-test, resulting in considerably more high-confidence identifications of rhythmic transcripts than previously described. Finally, to extend the utility of these data sets, a Web-based resource has been constructed (at http://wasabi.itmat.upenn.edu/circa/mouse ) that is freely available to the research community.
在视交叉上核(SCN)和外周组织中,昼夜节律振荡器驱动下游靶基因的节律性转录。最近,许多研究使用DNA微阵列系统地鉴定植物、果蝇、大鼠和小鼠中的振荡转录本。这些研究通过在1、2天或更长时间内每4小时对组织进行一次采样,鉴定出了几十到数百个有节律表达的基因。为了扩展这项工作,我们对从小鼠垂体中提取的RNA进行了DNA微阵列分析,每小时采样一次,共持续2天。使用COSOPT和Fisher G检验,错误发现率低于5%,以鉴定垂体中超过250个以24小时周期振荡的基因。我们发现,在昼夜节律日内增加采样频率显著提高了COSOPT和Fisher G检验的统计功效,从而比之前描述的方法能鉴定出更多高可信度的节律性转录本。最后,为了扩展这些数据集的实用性,构建了一个基于网络的资源库(网址为http://wasabi.itmat.upenn.edu/circa/mouse ),供研究界免费使用。