Ramsey Kathryn Moynihan, Affinati Alison H, Peek Clara B, Marcheva Biliana, Hong Hee-Kyung, Bass Joseph
Division of Endocrinology, Metabolism and Molecular Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Methods Mol Biol. 2013;1077:285-302. doi: 10.1007/978-1-62703-637-5_19.
Many of our behavioral and physiological processes display daily oscillations that are under the control of the circadian clock. The core molecular clock network is present in both the brain and peripheral tissues and is composed of a complex series of interlocking transcriptional/translational feedback loops that oscillate with a periodicity of ~24 h. Recent evidence has implicated NAD(+) biosynthesis and the sirtuin family of NAD(+)-dependent protein deacetylases as part of a novel feedback loop within the core clock network, findings which underscore the importance of taking circadian timing into consideration when designing and interpreting metabolic studies, particularly in regard to sirtuin biology. Thus, this chapter introduces both in vivo and in vitro circadian methods to analyze various sirtuin-related endpoints across the light-dark cycle and discusses the transcriptional, biochemical, and physiological outputs of the clock.
我们的许多行为和生理过程都表现出受昼夜节律时钟控制的每日振荡。核心分子时钟网络存在于大脑和外周组织中,由一系列复杂的相互连锁的转录/翻译反馈环组成,这些反馈环以约24小时的周期振荡。最近的证据表明,NAD(+)生物合成和依赖NAD(+)的蛋白质脱乙酰酶的沉默调节蛋白家族是核心时钟网络内一个新的反馈环的一部分,这些发现强调了在设计和解释代谢研究时考虑昼夜节律时间的重要性,特别是在沉默调节蛋白生物学方面。因此,本章介绍了体内和体外昼夜节律方法,以分析在明暗周期中各种与沉默调节蛋白相关的终点,并讨论了时钟的转录、生化和生理输出。