鳞翅目昆虫生物钟的研究进展与展望

经过长期的自然演化,昆虫生命活动和行为中有着明显的昼夜和季节的节律性周期变化——生物钟(circadian clock)。鳞翅目昆虫是当前物种最丰富的昆虫群体,包括蝴蝶和飞蛾约16万种,其中近70%的重要农林害虫属于鳞翅目。而鳞翅目昆虫生物钟的研究进展对于深入剖析鳞翅目昆虫丰富多变的生理行为调控机制,有效开展鳞翅目经济昆虫的生产及农林害虫防治具有重要意义。总结了昼夜节律生物钟对鳞翅目昆虫孵化与取食、生长与变态、生殖与滞育、求偶与迁飞等生理行为的影响,重点概述了鳞翅目昆虫生物钟分子调控机制、生物钟与内分泌激素协同调控机制等重要研究进展,并基于生物钟原理探讨了鳞翅目昆虫与植物协同进化关系及重要生态意义,展望了将生物钟理论应用于农业害虫防治和经济昆虫饲养改良的应用前景。     

Molecular mechanisms of the biological clock in cultured fibroblasts

In mammals, the central circadian pacemaker resides in the hypothalamic suprachiasmatic nucleus (SCN), but circadian oscillators also exist in peripheral tissues. Here, using wild-type and cryptochrome (mCry)-deficient cell lines derived from mCry mutant mice, we show that the peripheral oscillator in cultured fibroblasts is identical to the oscillator in the SCN in (i) temporal expression profiles of all known clock genes, (ii) the phase of the various mRNA rhythms (i.e., antiphase oscillation of Bmal1 and mPer genes), (iii) the delay between maximum mRNA levels and appearance of nuclear mPER1 and mPER2 protein, (iv) the inability to produce oscillations in the absence of functional mCry genes, and (v) the control of period length by mCRY proteins.     

哺乳动物生物钟昼夜节律同步机制的研究进展

哺乳动物昼夜节律生物钟是以24 h为周期的自主维持的振荡器。在分子水平上,生物钟的振荡由自身调控反馈环路的转录和翻译组成,并接受外界环境因素的影响,通过下丘脑视叉上核(Supra Ch iasm atic N u-cleus,SCN)中枢震荡器的同步整和而产生作用。文章对生物钟上下游的分布、反馈环路的转录、翻译后事件、视觉通路、钟的整和等方面进行了综述,并对其今后的研究方向作了展望。     

JETLAG resets the Drosophila circadian clock by promoting light-induced degradation of TIMELESS

Organisms ranging from bacteria to humans synchronize their internal clocks to daily cycles of light and dark. Photic entrainment of the Drosophila clock is mediated by proteasomal degradation of the clock protein TIMELESS (TIM). We have identified mutations in jetlag-a gene coding for an F-box protein with leucine-rich repeats-that result in reduced light sensitivity of the circadian clock. Mutant flies show rhythmic behavior in constant light, reduced phase shifts in response to light pulses, and reduced light-dependent degradation of TIM. Expression of JET along with the circadian photoreceptor cryptochrome (CRY) in cultured S2R cells confers light-dependent degradation onto TIM, thereby reconstituting the acute response + of the circadian clock to light in a cell culture system. Our results suggest that JET is essential for resetting the clock by transmitting light signals from CRY to TIM.     

The Neurospora checkpoint kinase 2: a regulatory link between the circadian and cell cycles

The clock gene period-4 (prd-4) in Neurospora was identified by a single allele displaying shortened circadian period and altered temperature compensation. Positional cloning followed by functional tests show that PRD-4 is an ortholog of mammalian checkpoint kinase 2 (Chk2). Expression of prd-4 is regulated by the circadian clock and, reciprocally, PRD-4 physically interacts with the clock component FRQ, promoting its phosphorylation. DNA-damaging agents can reset the clock in a manner that depends on time of day, and this resetting is dependent on PRD-4. Thus, prd-4, the Neurospora Chk2, identifies a molecular link that feeds back conditionally from circadian output to input and the cell cycle.     

The pseudo-response regulator Ppd-H1 provides adaptation to photoperiod in barley

Plants commonly use photoperiod (day length) to control the timing of flowering during the year, and variation in photoperiod response has been selected in many crops to provide adaptation to different environments and farming practices. Positional cloning identified Ppd-H1, the major determinant of barley photoperiod response, as a pseudo-response regulator, a class of genes involved in circadian clock function. Reduced photoperiod responsiveness of the ppd-H1 mutant, which is highly advantageous in spring-sown varieties, is explained by altered circadian expression of the photoperiod pathway gene CONSTANS and reduced expression of its downstream target, FT, a key regulator of flowering.     

基于Diurnal在线软件分析拟南芥生物节律基因对GABA的响应

Diurnal是一个基于网络的在线分析工具,利用该工具可以得到几种模式植物基因组范围内的昼夜和生物钟节律基因表达的变化规律。这一在线分析软件是一个可供搜索的数据库,它提供了几个基于网络的界面友好的数据挖掘工具,其结果以一种易于理解的方式呈现。在该研究中首先介绍了这个在线软件的使用方法,然后分析了几个拟南芥生物钟节律基因和开花基因在长日照条件下的表达模型,这些结果可以以图形和数字的方式下载用于后续的特定研究。结合在线数据,该研究表明,γ-氨基丁酸（GABA在1.0mmol/L）可以改变3个生物钟核心节律基因（TOC1,CCA1和LHY）的表达模式,这种模式的改变主要是增加了这些基因表达的幅度。     

Circadian clocks in daily and seasonal control of development

The rotation of the earth results in regular changes in the light environment, and organisms have evolved a molecular oscillator that allows them to anticipate these changes. This daily molecular oscillator, known as the circadian clock, regulates a diverse array of physiologies across a wide variety of organisms. This review highlights a few of the insights we have into circadian clock regulation of development, in both plants and animals. A common thread linking plants and animals is the use of the circadian clock to sense changes in day length and to mediate a diverse number of photoperiodic responses.     

Plant circadian clocks increase photosynthesis, growth, survival, and competitive advantage

Circadian clocks are believed to confer an advantage to plants, but the nature of that advantage has been unknown. We show that a substantial photosynthetic advantage is conferred by correct matching of the circadian clock period with that of the external light-dark cycle. In wild type and in long- and short-circadian period mutants of Arabidopsis thaliana, plants with a clock period matched to the environment contain more chlorophyll, fix more carbon, grow faster, and survive better than plants with circadian periods differing from their environment. This explains why plants gain advantage from circadian control.     

Identification of small molecule activators of cryptochrome

Impairment of the circadian clock has been associated with numerous disorders, including metabolic disease. Although small molecules that modulate clock function might offer therapeutic approaches to such diseases, only a few compounds have been identified that selectively target core clock proteins. From an unbiased cell-based circadian phenotypic screen, we identified KL001, a small molecule that specifically interacts with cryptochrome (CRY). KL001 prevented ubiquitin-dependent degradation of CRY, resulting in lengthening of the circadian period. In combination with mathematical modeling, our studies using KL001 revealed that CRY1 and CRY2 share a similar functional role in the period regulation. Furthermore, KL001-mediated CRY stabilization inhibited glucagon-induced gluconeogenesis in primary hepatocytes. KL001 thus provides a tool to study the regulation of CRY-dependent physiology and aid development of clock-based therapeutics of diabetes.     

Natural free-running period in vertebrate animal populations

Regression analysis (analysis of covariance) is contrasted with the conventional "mean period length" for estimating the length of period of the spontaneous activity frequency (free-running period) in population samples of Gila monsters (Heloderma suspectum) and kangaroo rats (Dipodomys merriami) in the Sonoran Desert. The mean period length in each population does not differ significantly from 24:00 hours (P > .05) and it does not differ significantly (P > .05) between the species studied; the probability that the free-running period a(in constant dark) in natural populations of Gila monsters and kangaroo rats is different from 24:00.0 is less than 1 in 1000 (P < .001). The so-called "mean period length" is of little or no use for precise determination of the period and phase relationships in circadian rhythms; moreover, it is entirely without value for statistical testing of differences either within or between populations.     

Antiphase oscillation of the left and right suprachiasmatic nuclei

An unusual property of the circadian timekeeping systems of animals is rhythm "splitting," in which a single daily period of physical activity (usually measured as wheel running) dissociates into two stably coupled components about 12 hours apart; this behavior has been ascribed to a clock composed of two circadian oscillators cycling in antiphase. We analyzed gene expression in the hypothalamic circadian clock, the suprachiasmatic nucleus (SCN), of behaviorally "split" hamsters housed in constant light. The results show that the two oscillators underlying the split condition correspond to the left and right sides of the bilaterally paired SCN.     

An hPer2 phosphorylation site mutation in familial advanced sleep phase syndrome

Familial advanced sleep phase syndrome (FASPS) is an autosomal dominant circadian rhythm variant; affected individuals are "morning larks" with a 4-hour advance of the sleep, temperature, and melatonin rhythms. Here we report localization of the FASPS gene near the telomere of chromosome 2q. A strong candidate gene (hPer2), a human homolog of the period gene in Drosophila, maps to the same locus. Affected individuals have a serine to glycine mutation within the casein kinase Iepsilon (CKIepsilon) binding region of hPER2, which causes hypophosphorylation by CKIepsilon in vitro. Thus, a variant in human sleep behavior can be attributed to a missense mutation in a clock component, hPER2, which alters the circadian period.     

Resetting of circadian time in peripheral tissues by glucocorticoid signaling

In mammals, circadian oscillators reside not only in the suprachiasmatic nucleus of the brain, which harbors the central pacemaker, but also in most peripheral tissues. Here, we show that the glucocorticoid hormone analog dexamethasone induces circadian gene expression in cultured rat-1 fibroblasts and transiently changes the phase of circadian gene expression in liver, kidney, and heart. However, dexamethasone does not affect cyclic gene expression in neurons of the suprachiasmatic nucleus. This enabled us to establish an apparent phase-shift response curve specifically for peripheral clocks in intact animals. In contrast to the central clock, circadian oscillators in peripheral tissues appear to remain responsive to phase resetting throughout the day.     

A circadian output in Drosophila mediated by neurofibromatosis-1 and Ras/MAPK

Output from the circadian clock controls rhythmic behavior through poorly understood mechanisms. In Drosophila, null mutations of the neurofibromatosis-1 (Nf1) gene produce abnormalities of circadian rhythms in locomotor activity. Mutant flies show normal oscillations of the clock genes period (per) and timeless (tim) and of their corresponding proteins, but altered oscillations and levels of a clock-controlled reporter. Mitogen-activated protein kinase (MAPK) activity is increased in Nf1 mutants, and the circadian phenotype is rescued by loss-of-function mutations in the Ras/MAPK pathway. Thus, Nf1 signals through Ras/MAPK in Drosophila. Immunohistochemical staining revealed a circadian oscillation of phospho-MAPK in the vicinity of nerve terminals containing pigment-dispersing factor (PDF), a secreted output from clock cells, suggesting a coupling of PDF to Ras/MAPK signaling.