The network of time: understanding the molecular circadian system

The circadian clock provides a temporal structure that modulates biological functions from the level of gene expression to performance and behaviour. Pioneering work on the fruitfly Drosophila has provided a basis for understanding how the temporal sequence of daily events is controlled in mammals. New insights have come from work on mammals, specifically from studying the daily activity profiles of clock mutant mice; from more detailed recordings of clock gene expression under different experimental conditions and in different tissues; and from the discovery and analysis of a growing number of additional clock genes. These new results are moving the model paradigm away from a simple negative feedback loop to a molecular network. Understanding the coupling and interactions of this network will help us to understand the evolution of the circadian system, advance medical diagnosis and treatment, improve the health of shift workers and frequent travellers, and will generally enable the treatment of clock-related pathologies.     

果蝇昼夜节律的分子机制研究进展

果蝇由于遗传易操作性而成为一个研究昼夜节律分子机制的理想模式生物 . 到目前为止,通过遗传学和生物化学方法已经鉴定到 10 多个时钟基因 (clock genes) 和许多时钟相关基因,包括时钟输入基因和钟控基因 . 这些时钟基因以及它们的相应产物组成两个互相依赖的转录 / 翻译反馈环路,从而调节行为和生理的昼夜节律 . 果蝇这种核心钟的工作原理同样见于哺乳动物 .     

Effects of Different PER Translational Kinetics on the Dynamics of a Core Circadian Clock Model

Living beings display self-sustained daily rhythms in multiple biological processes, which persist in the absence of external cues since they are generated by endogenous circadian clocks. The period (per) gene is a central player within the core molecular mechanism for keeping circadian time in most animals. Recently, the modulation PER translation has been reported, both in mammals and flies, suggesting that translational regulation of clock components is important for the proper clock gene expression and molecular clock performance. Because translational regulation ultimately implies changes in the kinetics of translation and, therefore, in the circadian clock dynamics, we sought to study how and to what extent the molecular clock dynamics is affected by the kinetics of PER translation. With this objective, we used a minimal mathematical model of the molecular circadian clock to qualitatively characterize the dynamical changes derived from kinetically different PER translational mechanisms. We found that the emergence of self-sustained oscillations with characteristic period, amplitude, and phase lag (time delays) between per mRNA and protein expression depends on the kinetic parameters related to PER translation. Interestingly, under certain conditions, a PER translation mechanism with saturable kinetics introduces longer time delays than a mechanism ruled by a first-order kinetics. In addition, the kinetic laws of PER translation significantly changed the sensitivity of our model to parameters related to the synthesis and degradation of per mRNA and PER degradation. Lastly, we found a set of parameters, with realistic values, for which our model reproduces some experimental results reported recently for Drosophila melanogaster and we present some predictions derived from our analysis.     

Timeless与生物钟基因

综述了timeless基因的发现、多态性和重要功能。timeless是最先被发现的两个生物钟基因之一。生物钟的昼夜节律由PER、TIM、CLOCK和CYCLE4个生物钟齿轮组成的正负反馈回路进行调节。其中TIM可以受光因子调控,它还可以与PER形成异二聚体,通过正负调控方式调节果蝇的昼夜节律行为。     

The 2006 Pittendrigh/Aschoff lecture: new roles for old proteins in the Drosophila circadian clock

Circadian behaviors in the animal kingdom are regulated by a small set of conserved genes. Starting with a historical perspective focused on Drosophila, the authors describe how the recurrent discovery of circadian clock genes uncovered a molecular mechanism associated with cycling gene expression. These molecular cycles appear to emerge from delayed negative and positive feedback. The authors will then introduce a novel timing mechanism uncovered by a single cell-based assay, with the new ideas and prospects for future research that it has raised.     

Setting the clock--by nature: circadian rhythm in the fruitfly Drosophila melanogaster

Nowadays humans mainly rely on external, unnatural clocks such as of cell phones and alarm clocks--driven by circuit boards and electricity. Nevertheless, our body is under the control of another timer firmly anchored in our genes. This evolutionary very old biological clock drives most of our physiology and behavior. The genes that control our internal clock are conserved among most living beings. One organism that shares this ancient clock mechanism with us humans is the fruitfly Drosophila melanogaster. Since it turned out that Drosophila is an excellent model, it is no surprise that its clock is very well and intensely investigated. In the following review we want to display an overview of the current understanding of Drosophila's circadian clock.     

Flies, clocks and evolution.

The negative feedback model for gene regulation of the circadian mechanism is described for the fruitfly, Drosophila melanogaster. The conservation of function of clock molecules is illustrated by comparison with the mammalian circadian system, and the apparent swapping of roles between various canonical clock gene components is highlighted. The role of clock gene duplications and divergence of function is introduced via the timeless gene. The impressive similarities in clock gene regulation between flies and mammals could suggest that variation between more closely related species within insects might be minimal. However, this is not borne out because the expression of clock molecules in the brain of the giant silk moth, Antheraea pernyi, is not easy to reconcile with the negative feedback roles of the period and timeless genes. Variation in clock gene sequences between and within fly species is examined and the role of co-evolution between and within clock molecules is described, particularly with reference to adaptive functions of the circadian phenotype.     

The Drosophila circadian clock: what we know and what we don't know.

Circadian rhythms are regulated by endogenous body clocks, which are formed by rhythmic cycles of clock gene expression. Almost all reviews of the Drosophila circadian clock state that the intracellular oscillator is based on a simple negative feedback loop. However, not many 'simple' feedback loops in biology last for 24 h. Instead, the Drosophila clock is a series of precisely timed steps that are deliberately slow. In this paper, I will discuss the current model for how the Drosophila clock is regulated, and ask what questions remain to be answered.     

Circadian clocks: A cry in the dark?

Cryptochrome proteins are key components of the circadian systems of both Drosophila and mammals. In Drosophila, they appear to be responsible for the entrainment of the circadian clock by the light-dark cycle, while in mammals they perform an important role in rhythm generation itself.     

How does the circadian clock send timing information to the brain?

This paper discusses circadian output in terms of the signaling mechanisms used by circadian pacemaker neurons. In mammals, the suprachiasmatic nucleus houses a clock controlling several rhythmic events. This nucleus contains one or more pacemaker circuits, and exhibits diversity in transmitter content and in axonal projections. In Drosophila, a comparable circadian clock is located among period -expressing neurons, a sub-set of which (called LN-vs) express the neuropeptide PDF. Genetic experiments indicate LN-vs are the primary pacemakers neurons controlling daily locomotion and that PDF is the principal circadian transmitter. Further definition of pacemaker properties in several model systems will provide a useful basis with which to describe circadian output mechanisms.     

生物钟机制研究进展

由生物体内源性生物钟所产生的昼夜节律是近年来生命科学的研究热点之一。几种模型生物（蓝细菌、脉孢菌、拟南芥、果蝇、小鼠）的生物钟相关基因相继被克隆和鉴定,为理解昼夜节律的分子机制奠定了基础。振荡器蛋白对其编码基因的负反馈调控可能是不同生物的生物运作普遍机制,在此基础上,不同生物有不尽相同的调控方式;隐色素可能是高等生物的共同生物钟光受体。     

Circadian clocks in the mammalian brain

Daily cycles in physiology and behaviour are probably a universal feature of multicellular organisms. These rhythms are predominantly driven by endogenous clocks with a periodicity approximating to one day, i.e. circadian. In mammals, the circadian clock governing activity/ rest, neuroendocrine and autonomic rhythms lies in the hypothalamus, in the suprachiasmatic nuclei (SCN). Intrinsic circadian oscillators are also present in the retina. The SCN "clockwork" is based on a cell autonomous, genetically determined mechanism. Mammalian homologues of a number of Drosophila genes which encode elements of the fly circadian mechanism have recently been identified. In Drosophila, the protein products of these genes interact in a negative feedback loop, establishing a circadian cycle in gene expression. Characterisation of the roles played by putative mammalian clock genes in the SCN, and how the emergent cellular signal imposes order over the entire neuraxis, will provide a fundamental contribution to our understanding of the molecular basis of behaviour. BioEssays 22:23-31, 2000.     

Altered behavioral rhythms and clock gene expression in mice with a targeted mutation in the Period1 gene

A group of specialized genes has been defined to govern the molecular mechanisms controlling the circadian clock in mammals. Their expression and the interactions among their products dictate circadian rhythmicity. Three genes homologous to Drosophila period exist in the mouse and are thought to be major players in the biological clock. Here we present the generation of mice in which the founding member of the family, Per1, has been inactivated by homologous recombination. These mice present rhythmicity in locomotor activity, but with a period almost 1 h shorter than wild-type littermates. Moreover, the expression of clock genes in peripheral tissues appears to be delayed in Per1 mutant animals. Importantly, light-induced phase shifting appears conserved. The oscillatory expression of clock genes and the induction of immediate-early genes in response to light in the master clock structure, the suprachiasmatic nucleus, are unaffected. Altogether, these data demonstrate that Per1 plays a distinct role within the Per family, as it may be involved predominantly in peripheral clocks and/or in the output pathways of the circadian clock.     

生物钟基因period的分子生物学

生物过程的昼夜节律是所有真核生物和部分原核生物的基本特征。自从ｐｅｒｉｏｄ基因被克隆以后,生物钟基因的研究已经取得长足的促进。本文回顾了ｐｅｒ基因及生物钟分子机制的研究历史,旨在为人类复杂行为的研究提供一条思路。     

Neural circuits underlying circadian behavior in Drosophila melanogaster

Circadian clocks include control systems for organizing daily behavior. Such a system consists of a time-keeping mechanism (the clock or pacemaker), input pathways for entraining the clock, and output pathways for producing overt rhythms in behavior and physiology. In Drosophila melanogaster, as in mammals, neural circuits play vital roles in all three functional subdivisions of the circadian system. Regarding the pacemaker, multiple clock neurons, each with cell-autonomous pacemaker capability, are coupled to each other in a network. The outputs of different sets of clock neurons in this network combine to produce the normal bimodal pattern of locomotor activity observed in Drosophila. Regarding input, multiple sensory modalities (including light, temperature, and pheromones) use their own circuitry to entrain the clock. Regarding output, distinct circuits are likely involved for controlling the timing of eclosion and for generating the locomotor activity rhythms. This review summarizes work on all of these circadian circuits, and discusses the broader utility of studying the fly's circadian system.