Martha Merrow

Life between Clocks: Daily Temporal Patterns of Human Chronotypes:

Human behavior shows large interindividual variation in temporal organization. Extreme “larks” wake up when extreme “owls” fall asleep. These chronotypes are attributed to differences in the circadian clock, and in animals, the genetic basis of similar phenotypic differences is well established. To better understand the genetic basis of temporal organization in humans, the authors developed a questionnaire to document individual sleep times, self-reported light exposure, and self-assessed chronotype, considering work and free days separately. This report summarizes the results of 500 questionnaires completed in a pilot study. Individual sleep times show large differences between work and free days, except for extreme early types. During the workweek, late chronotypes accumulate considerable sleep debt, for which they compensate on free days by lengthening their sleep by several hours. For all chronotypes, the amount of time spent outdoors in broad daylight significantly affects the timing of sleep: Increased self-reported light exposure advances sleep. The timing of self-selected sleep is multifactorial, including genetic disposition, sleep debt accumulated on workdays, and light exposure. Thus, accurate assessment of genetic chronotypes has to incorporate all of these parameters. The dependence of human chronotype on light, that is, on the amplitude of the light:dark signal, follows the known characteristics of circadian systems in all other experimental organisms. Our results predict that the timing of sleep has changed during industrialization and that a majority of humans are sleep deprived during the workweek. The implications are far ranging concerning learning, memory, vigilance, performance, and quality of life.

Social jetlag: Misalignment of biological and social time.

Humans show large differences in the preferred timing of their sleep and activity. This so‐called “chronotype” is largely regulated by the circadian clock. Both genetic variations in clock genes and environmental influences contribute to the distribution of chronotypes in a given population, ranging from extreme early types to extreme late types with the majority falling between these extremes. Social (e.g., school and work) schedules interfere considerably with individual sleep preferences in the majority of the population. Late chronotypes show the largest differences in sleep timing between work and free days leading to a considerable sleep debt on work days, for which they compensate on free days. The discrepancy between work and free days, between social and biological time, can be described as ‘social jetlag.’ Here, we explore how sleep quality and psychological wellbeing are associated with individual chronotype and/or social jetlag. A total of 501 volunteers filled out the Munich ChronoType Questionnaire (MCTQ) as well as additional questionnaires on: (i) sleep quality (SF‐A), (ii) current psychological wellbeing (Basler Befindlichkeitsbogen), (iii) retrospective psychological wellbeing over the past week (POMS), and (iv) consumption of stimulants (e.g., caffeine, nicotine, and alcohol). Associations of chronotype, wellbeing, and stimulant consumption are strongest in teenagers and young adults up to age 25 yrs. The most striking correlation exists between chronotype and smoking, which is significantly higher in late chronotypes of all ages (except for those in retirement). We show these correlations are most probably a consequence of social jetlag, i.e., the discrepancies between social and biological timing rather than a simple association to different chronotypes. Our results strongly suggest that work (and school) schedules should be adapted to chronotype whenever possible.

Peroxiredoxins are conserved markers of circadian rhythms

Cellular life emerged ∼3.7 billion years ago. With scant exception, terrestrial organisms have evolved under predictable daily cycles owing to the Earth’s rotation. The advantage conferred on organisms that anticipate such environmental cycles has driven the evolution of endogenous circadian rhythms that tune internal physiology to external conditions. The molecular phylogeny of mechanisms driving these rhythms has been difficult to dissect because identified clock genes and proteins are not conserved across the domains of life: Bacteria, Archaea and Eukaryota. Here we show that oxidation–reduction cycles of peroxiredoxin proteins constitute a universal marker for circadian rhythms in all domains of life, by characterizing their oscillations in a variety of model organisms. Furthermore, we explore the interconnectivity between these metabolic cycles and transcription–translation feedback loops of the clockwork in each system. Our results suggest an intimate co-evolution of cellular timekeeping with redox homeostatic mechanisms after the Great Oxidation Event ∼2.5 billion years ago.

The human circadian clock entrains to sun time

Document S1. Supplemental Experimental Procedure and One FigurexDownload (.05 MB ) Document S1. Supplemental Experimental Procedure and One Figure

The Art of Entrainment

The circadian system actively synchronizes the temporal sequence of biological functions with the environment. The oscillatory behavior of the system ensures that entrainment is not passive or driven and therefore allows for great plasticity and adaptive potential. With the tools at hand, we now can concentrate on the most important circadian question: How is the complex task of entrainment achieved by anatomical, cellular, and molecular components? Understanding entrainment is equal to understanding the circadian system. The results of this basic research will help us to understand temporal ecology and will allow us to improve conditions for humans in industrialized societies.

Assignment of circadian function for the Neurospora clock gene frequency

Circadian clocks consist of three elements: entrainment pathways (inputs), the mechanism generating the rhythmicity (oscillator), and the output pathways that control the circadian rhythms. It is difficult to assign molecular clock components to any one of these elements. Experiments show that inputs can be circadianly regulated and outputs can feed back on the oscillator,. Mathematical simulations indicate that under- or overexpression of a gene product can result in arrhythmicity, whether the protein is part of the oscillator or substantially part of a rhythmically expressed input pathway. To distinguish between these two possibilities, we used traditional circadian entrainment protocols, on a genetic model system, Neurospora crassa.

The human circadian clock's seasonal adjustment is disrupted by daylight saving time

A quarter of the worlds population is subjected to a 1 hr time change twice a year (daylight saving time, DST). This reflects a change in social clocks, not environmental ones (e.g., dawn). The impact of DST is poorly understood. Circadian clocks use daylight to synchronize (entrain) to the organisms environment. Entrainment is so exact that humans adjust to the east-west progression of dawn within a given time zone. In a large survey (n = 55,000), we show that the timing of sleep on free days follows the seasonal progression of dawn under standard time, but not under DST. In a second study, we analyzed the timing of sleep and activity for 8 weeks around each DST transition in 50 subjects who were chronotyped (analyzed for their individual phase of entrainment). Both parameters readily adjust to the release from DST in autumn but the timing of activity does not adjust to the DST imposition in spring, especially in late chronotypes. Our data indicate that the human circadian system does not adjust to DST and that its seasonal adaptation to the changing photoperiods is disrupted by the introduction of summer time. This disruption may extend to other aspects of seasonal biology in humans.

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.

Entrainment of the Human Circadian Clock

Humans are an excellent model system for studying entrainment of the circadian clock in the real world. Unlike the situation in laboratory experiments, entrainment under natural conditions is achieved by different external signals as well as by internal signals generated by multiple feedbacks within the system (e.g., behavior-dependent light and temperature changes, melatonin levels, or regular nutrient intake). Signals that by themselves would not be sufficient zeitgebers may contribute to entrainment in conjunction with other self-sufficient zeitgeber signals (e.g., light). The investigation of these complex zeitgeber interactions seems to be problematic in most model systems and strengthens the human system for circadian research. Here, we review our endeavors measuring human entrainment in real life, predominantly with the help of the Munich ChronoType Questionnaire (MCTQ). The large number of participants in our current MCTQ database allows accurate quantification of the human phase of entrainment (chronotype) and how it depends on age or sex. We also present new data showing how chronotype depends on natural light exposure. The results indicate the importance of zeitgeber strength on human entrainment and help in understanding the differences in chronotype, e.g., between urban and rural regions.

A PEST-like element in FREQUENCY determines the length of the circadian period in Neurospora crassa

FREQUENCY (FRQ) is a crucial element of the circadian clock in Neurospora crassa. In the course of a circadian day FRQ is successively phosphorylated and degraded. Here we report that two PEST‐like elements in FRQ, PEST‐1 and PEST‐2, are phosphorylated in vitro by recombinant CK‐1a and CK‐1b, two newly identified Neurospora homologs of casein kinase 1ϵ. CK‐1a is localized in the cytosol and the nuclei of Neurospora and it is in a complex with FRQ in vivo. Deletion of PEST‐1 results in hypophosphorylation of FRQ and causes significantly increased protein stability. A strain harboring the mutant frqΔPEST‐1 gene shows no rhythmic conidiation. Despite the lack of overt rhythmicity, frqΔPEST‐1 RNA and FRQΔPEST‐1 protein are rhythmically expressed and oscillate in constant darkness with a circadian period of 28 h. Thus, by deletion of PEST‐1 the circadian period is lengthened and overt rhythmicity is dissociated from molecular oscillations of clock components.