Vincent van der Vinne

Cold and hunger induce diurnality in a nocturnal mammal

Significance The circadian system drives daily rhythms in physiology and behavior. Mammals in nature can change their daily activity rhythms, but causes and consequences of this behavioral plasticity are unknown. Here we show that nocturnal mice become diurnal when challenged by cold or hunger. Negative energy balance changes hormonal, physiological and behavioral rhythms without modifying the rhythm of the circadian pacemaker in the suprachiasmatic nucleus. This response is adaptive because activity during daytime warmth and resting in a buffered environment during the cold nighttime generally reduces energy expenditure. This mechanism may explain why nighttime activity in humans generally evokes higher energy uptake and subsequent obesity and metabolic syndrome, as seen in late chronotypes and night shift workers. The mammalian circadian system synchronizes daily timing of activity and rest with the environmental light–dark cycle. Although the underlying molecular oscillatory mechanism is well studied, factors that influence phenotypic plasticity in daily activity patterns (temporal niche switching, chronotype) are presently unknown. Molecular evidence suggests that metabolism may influence the circadian molecular clock, but evidence at the level of the organism is lacking. Here we show that a metabolic challenge by cold and hunger induces diurnality in otherwise nocturnal mice. Lowering ambient temperature changes the phase of circadian light–dark entrainment in mice by increasing daytime and decreasing nighttime activity. This effect is further enhanced by simulated food shortage, which identifies metabolic balance as the underlying common factor influencing circadian organization. Clock gene expression analysis shows that the underlying neuronal mechanism is downstream from or parallel to the main circadian pacemaker (the hypothalamic suprachiasmatic nucleus) and that the behavioral phenotype is accompanied by phase adjustment of peripheral tissues. These findings indicate that nocturnal mammals can display considerable plasticity in circadian organization and may adopt a diurnal phenotype when energetically challenged. Our previously defined circadian thermoenergetics hypothesis proposes that such circadian plasticity, which naturally occurs in nocturnal mammals, reflects adaptive maintenance of energy balance. Quantification of energy expenditure shows that diurnality under natural conditions reduces thermoregulatory costs in small burrowing mammals like mice. Metabolic feedback on circadian organization thus provides functional benefits by reducing energy expenditure. Our findings may help to clarify relationships between sleep–wake patterns and metabolic phenotypes in humans.

Ambient temperature shapes reproductive output during pregnancy and lactation in the common vole (Microtus arvalis): a test of the heat dissipation limit theory

SUMMARY The heat dissipation limit theory suggests that heat generated during metabolism limits energy intake and, thus, reproductive output. Experiments in laboratory strains of mice and rats, and also domestic livestock generally support this theory. Selection for many generations in the laboratory and in livestock has increased litter size or productivity in these animals. To test the wider validity of the heat dissipation limit theory, we studied common voles (Microtus arvalis), which have small litter sizes by comparison with mice and rats, and regular addition of wild-caught individuals of this species to our laboratory colony ensures a natural genetic background. A crossover design of ambient temperatures (21 and 30°C) during pregnancy and lactation was used. High ambient temperature during lactation decreased milk production, slowing pup growth. The effect on pup growth was amplified when ambient temperature was also high during pregnancy. Shaving fur off dams at 30°C resulted in faster growth of pups; however, no significant increase in food intake and or milk production was detected. With increasing litter size (natural and enlarged), asymptotic food intake during lactation levelled off in the largest litters at both 21 and 30°C. Interestingly, the effects of lactation temperature on pup growth where also observed at smaller litter sizes. This suggests that vole dams trade-off costs associated with hyperthermia during lactation with the yield from investment in pup growth. Moreover, pup survival was higher at 30°C, despite lower growth, probably owing to thermoregulatory benefits. It remains to be seen how the balance is established between the negative effect of high ambient temperature on maternal milk production and pup growth (and/or future reproduction of the dam) and the positive effect of high temperatures on pup survival. This balance ultimately determines the effect of different ambient temperatures on reproductive success.

Timing of Examinations Affects School Performance Differently in Early and Late Chronotypes

Circadian clocks of adolescents typically run late—including sleep times—yet adolescents generally are expected at school early in the morning. Due to this mismatch between internal (circadian) and external (social) times, adolescents suffer from chronic sleep deficiency, which, in turn, affects academic performance negatively. This constellation affects students’ future career prospects. Our study correlates chronotype and examination performance. In total, 4734 grades were collected from 741 Dutch high school students (ages 11-18 years) who had completed the Munich ChronoType Questionnaire to estimate their internal time. Overall, the lowest grades were obtained by students who were very late chronotypes (MSFsc > 5.31 h) or slept very short on schooldays (SDw < 7.03 h). The effect of chronotype on examination performance depended on the time of day that examinations were taken. Opposed to late types, early chronotypes obtained significantly higher grades during the early (0815-0945 h) and late (1000-1215 h) morning. This group difference in grades disappeared in the early afternoon (1245-1500 h). Late types also obtained lower grades than early types when tested at the same internal time (hours after MSFsc), which may reflect general attention and learning disadvantages of late chronotypes during the early morning. Our results support delaying high school starting times as well as scheduling examinations in the early afternoon to avoid discrimination of late chronotypes and to give all high school students equal academic opportunities.

Strong pituitary and hypothalamic responses to photoperiod but not to 6-methoxy-2-benzoxazolinone in female common voles (Microtus arvalis)

The annual cycle of changing day length (photoperiod) is widely used by animals to synchronise their biology to environmental seasonality. In mammals, melatonin is the key hormonal relay for the photoperiodic message, governing thyroid-stimulating hormone (TSH) production in the pars tuberalis (PT) of the pituitary stalk. TSH acts on neighbouring hypothalamic cells known as tanycytes, which in turn control hypothalamic function through effects on thyroid hormone (TH) signalling, mediated by changes in expression of the type II and III deiodinases (Dio2 and Dio3, respectively). Among seasonally breeding rodents, voles of the genus Microtus are notable for a high degree of sensitivity to nutritional and social cues, which act in concert with photoperiod to control reproductive status. In the present study, we investigated whether the TSH/Dio2/Dio3 signalling pathway of female common voles (Microtus arvalis) shows a similar degree of photoperiodic sensitivity to that described in other seasonal mammal species. Additionally, we sought to determine whether the plant metabolite 6-methoxy-2-benzoxazolinone (6-MBOA), described previously as promoting reproductive activation in voles, had any influence on the TSH/Dio2/Dio3 system. Our data demonstrate a high degree of photoperiodic sensitivity in this species, with no observable effects of 6-MBOA on upstream pituitary/hypothalamic gene expression. Further studies are required to characterise how photoperiodic and nutritional signals interact to modulate hypothalamic TH signalling pathways in mammals.

Diurnality as an energy-saving strategy: energetic consequences of temporal niche switching in small mammals

ABSTRACT Endogenous daily (circadian) rhythms allow organisms to anticipate daily changes in the environment. Most mammals are specialized to be active during the night (nocturnal) or day (diurnal). However, typically nocturnal mammals become diurnal when energetically challenged by cold or hunger. The circadian thermo-energetics (CTE) hypothesis predicts that diurnal activity patterns reduce daily energy expenditure (DEE) compared with nocturnal activity patterns. Here, we tested the CTE hypothesis by quantifying the energetic consequences of relevant environmental factors in mice. Under natural conditions, diurnality reduces DEE by 6–10% in energetically challenged mice. Combined with night-time torpor, as observed in mice under prolonged food scarcity, DEE can be reduced by ∼20%. The dominant factor determining the energetic benefit of diurnality is thermal buffering provided by a sheltered resting location. Compared with nocturnal animals, diurnal animals encounter higher ambient temperatures during both day and night, leading to reduced thermogenesis costs in temperate climates. Analysis of weather station data shows that diurnality is energetically beneficial on almost all days of the year in a temperate climate region. Furthermore, diurnality provides energetic benefits at all investigated geographical locations on European longitudinal and latitudinal transects. The reduction of DEE by diurnality provides an ultimate explanation for temporal niche switching observed in typically nocturnal small mammals under energetically challenging conditions. Diurnality allows mammals to compensate for reductions in food availability and temperature as it reduces energetic needs. The optimal circadian organization of an animal ultimately depends on the balance between energetic consequences and other fitness consequences of the selected temporal niche. Highlighted Article: Computational modeling reveals energetic benefits for diurnal small mammals as predicted by the circadian thermo-energetics hypothesis.

Flexible clock systems : adjusting the temporal programme

Under natural conditions, many aspects of the abiotic and biotic environment vary with time of day, season or even era, while these conditions are typically kept constant in laboratory settings. The timing information contained within the environment serves as critical timing cues for the internal biological timing system, but how this system drives daily rhythms in behaviour and physiology may also depend on the internal state of the animal. The disparity between timing of these cues in natural and laboratory conditions can result in substantial differences in the scheduling of behaviour and physiology under these conditions. In nature, temporal coordination of biological processes is critical to maximize fitness because they optimize the balance between reproduction, foraging and predation risk. Here we focus on the role of peripheral circadian clocks, and the rhythms that they drive, in enabling adaptive phenotypes. We discuss how reproduction, endocrine activity and metabolism interact with peripheral clocks, and outline the complex phenotypes arising from changes in this system. We conclude that peripheral timing is critical to adaptive plasticity of circadian organization in the field, and that we must abandon standard laboratory conditions to understand the mechanisms that underlie this plasticity which maximizes fitness under natural conditions. This article is part of the themed issue ‘Wild clocks: integrating chronobiology and ecology to understand timekeeping in free-living animals’.

The flexible clock: Predictive and reactive homeostasis, energy balance and the circadian regulation of sleep-wake timing

ABSTRACT The Darwinian fitness of mammals living in a rhythmic environment depends on endogenous daily (circadian) rhythms in behavior and physiology. Here, we discuss the mechanisms underlying the circadian regulation of physiology and behavior in mammals. We also review recent efforts to understand circadian flexibility, such as how the phase of activity and rest is altered depending on the encountered environment. We explain why shifting activity to the day is an adaptive strategy to cope with energetic challenges and show how this can reduce thermoregulatory costs. A framework is provided to make predictions about the optimal timing of activity and rest of non-model species for a wide range of habitats. This Review illustrates how the timing of daily rhythms is reciprocally linked to energy homeostasis, and it highlights the importance of this link in understanding daily rhythms in physiology and behavior. Summary: The daily temporal niche of animals is adaptive and depends on environmental and physiological context.

Temporal niche switching and reduced nest attendance in response to heat dissipation limits in lactating common voles (Microtus arvalis)

According to the heat dissipation limit theory, maximum metabolic turnover is limited by the capacity of the body to dissipate excess heat. Small mammals, including common voles (Microtus arvalis), face a heat dissipation limitation during lactation. Pup growth and milk production are reduced under higher ambient temperatures. Heat dissipation problems might in part be alleviated by modifying behavior, such as reducing nest attendance and being active at energetically optimal times of day. According to the circadian thermo-energetics hypothesis, animals can make use of daily ambient temperature fluctuations to alter their energetic expenditure. In this study we test whether heat challenged (housing at 30 °C compared to 21 °C) lactating common voles allocate their time differently among behaviors and whether their ultradian and circadian behavioral rhythmicity are altered. Behavior was scored every 13 min from automated picture recordings, while general locomotor activity was measured by passive infrared detectors to assess ultradian and circadian organization. The effects of ambient temperature on the ultradian organization of behavior were assessed by determining the ultradian period length and the distribution of activity within the ultradian bout. Changes in circadian organization were assessed by the distribution of activity over the light and dark phase. As a complementary measure nest temperature recordings were used to quantify nest attendance distribution between day and night. Lactating dams at 30 °C reduced the fraction of time spent on the nest while increasing the fraction of time resting without pups away from the nest. The ultradian period of locomotor activity was longer in voles housed at 30 °C during pregnancy and lactation, but not after weaning when the pups were removed. No differences in the distribution of activity within the ultradian bout could be detected. The circadian organization was also modulated by ambient temperature. Lactating voles housed at 30 °C became more day active and a loss of day-night differences in nest temperature suggests a shift of nest attendance towards the night. Reducing the time attending the nest can reduce the risk of hyperthermia, and may be the behavioral component resulting in lower milk production and hence reproductive output. Becoming more day active allows feeding and nursing of the pups during the rest phase to occur during the night at which lower ambient temperatures are expected in the field. In natural situations this strategy will increase heat dissipation and lactation capacity. Whether there are similar benefits associated with a longer ultradian period is currently unknown, but these are likely to result from decreased energy turnover at 30 °C. In conclusion, our study shows that lactating common voles facing heat dissipation problems re-organize their behavior in a way that can maximize heat dissipation capabilities and thereby optimize lactation capacity.

Functionally Complete Excision of Conditional Alleles in the Mouse Suprachiasmatic Nucleus by Vgat-ires-Cre

Mice with targeted gene disruption have provided important information about the molecular mechanisms of circadian clock function. A full understanding of the roles of circadian-relevant genes requires manipulation of their expression in a tissue-specific manner, ideally including manipulation with high efficiency within the suprachiasmatic nuclei (SCN). To date, conditional manipulation of genes within the SCN has been difficult. In a previously developed mouse line, Cre recombinase was inserted into the vesicular GABA transporter (Vgat) locus. Since virtually all SCN neurons are GABAergic, this Vgat-Cre line seemed likely to have high efficiency at disrupting conditional alleles in SCN. To test this premise, the efficacy of Vgat-Cre in excising conditional (fl, for flanked by LoxP) alleles in the SCN was examined. Vgat-Cre-mediated excision of conditional alleles of Clock or Bmal1 led to loss of immunostaining for products of the targeted genes in the SCN. Vgat-Cre+; Clockfl/fl; Npas2m/m mice and Vgat-Cre+; Bmal1fl/fl mice became arrhythmic immediately upon exposure to constant darkness, as expected based on the phenotype of mice in which these genes are disrupted throughout the body. The phenotype of mice with other combinations of Vgat-Cre+, conditional Clock, and mutant Npas2 alleles also resembled the corresponding whole-body knockout mice. These data indicate that the Vgat-Cre line is useful for Cre-mediated recombination within the SCN, making it useful for Cre-enabled technologies including gene disruption, gene replacement, and opto- and chemogenetic manipulation of the SCN circadian clock.

Lower school performance in late chronotypes: underlying factors and mechanisms

Success at school determines future career opportunities. We described a time-of-day specific disparity in school performance between early and late chronotypes. Several studies showed that students with a late chronotype and short sleep duration obtain lower grades, suggesting that early school starting times handicap their performance. How chronotype, sleep duration, and time of day impact school performance is not clear. At a Dutch high school, we collected 40,890 grades obtained in a variety of school subjects over an entire school year. We found that the strength of the effect of chronotype on grades was similar to that of absenteeism, and that late chronotypes were more often absent. The difference in grades between the earliest 20% and the latest 20% of chronotypes corresponds to a drop from the 55th to 43rd percentile of grades. In academic subjects using mainly fluid cognition (scientific subjects), the correlation with grades and chronotype was significant while subjects relying on crystallised intelligence (humanistic/linguistic) showed no correlation with chronotype. Based on these and previous results, we can expand our earlier findings concerning exam times: students with a late chronotype are at a disadvantage in exams on scientific subjects, and when they are examined early in the day.