Food-entrainable Circadian Rhythms Research Articles

Enhanced food anticipatory activity associated with enhanced activation of extrahypothalamic neural pathways in serotonin2C receptor null mutant mice.

The ability to entrain circadian rhythms to food availability is important for survival. Food-entrained circadian rhythms are characterized by increased locomotor activity in anticipation of food availability (food anticipatory activity). However, the molecular components and neural circuitry underlying the regulation of food anticipatory activity remain unclear. Here we show that serotonin2C receptor (5-HT2CR) null mutant mice subjected to a daytime restricted feeding schedule exhibit enhanced food anticipatory activity compared to wild-type littermates, without phenotypic differences in the impact of restricted feeding on food consumption, body weight loss, or blood glucose levels. Moreover, we show that the enhanced food anticipatory activity in 5-HT2CR null mutant mice develops independent of external light cues and persists during two days of total food deprivation, indicating that food anticipatory activity in 5-HT2CR null mutant mice reflects the locomotor output of a food-entrainable oscillator. Whereas restricted feeding induces c-fos expression to a similar extent in hypothalamic nuclei of wild-type and null mutant animals, it produces enhanced expression in the nucleus accumbens and other extrahypothalamic regions of null mutant mice relative to wild-type subjects. These data suggest that 5-HT2CRs gate food anticipatory activity through mechanisms involving extrahypothalamic neural pathways.

The hunting of the snark. Interpretive issues in neurobiological analysis of food-entrained circadian rhythms

The hunting of the snark. Interpretive issues in neurobiological analysis of food-entrained circadian rhythms

Behavioral analysis of circadian rhythms: Entraining the circadian clock and determining the food-entrainable oscillator mechanism

Circadian rhythms are oscillations in behavior and physiological functions that are regulated by internal biological clocks. In mammals, the circadian rhythms can be best studied in the hypothalamic suprachiasmatic nucleus (SCN). The SCN has been considered as the master clock of circadian rhythms. SCN lesions abolish the circadian rhythms and transplantation of the SCN restores the behavioral rhythm; these findings confirm the role of the SCN as a circadian pacemaker. Recent discovery of molecular machineries of circadian oscillation enables us to examine genetically manipulated, molecular-clock-deficient mice, and use of these animal models has enhanced our understanding of biological clocks. It has been reported that prefeeding locomotor activity under a temporal restricted feeding condition exhibits the defining characteristic of a circadian rhythm. The prefeeding activity is driven by a distinct mechanism from the SCN. Despite extensive studies performed with the lesion strategy, the anatomic location of the food-entrainable oscillator has not been determined. Furthermore, recent studies using molecular-clock deficient mice reveal that the known circadian clock is dispensable for appearance of the prefeeding activity. Thus, examining the prefeeding activity alone is not sufficient and further examination for circadian oscillatory properties of the prefeeding activity rhythm is essential for understanding the food-entrainable circadian rhythms.

Standards of evidence in chronobiology: critical review of a report that restoration of Bmal1 expression in the dorsomedial hypothalamus is sufficient to restore circadian food anticipatory rhythms in Bmal1-/- mice

Daily feeding schedules generate food anticipatory rhythms of behavior and physiology that exhibit canonical properties of circadian clock control. The molecular mechanisms and location of food-entrainable circadian oscillators hypothesized to control food anticipatory rhythms are unknown. In 2008, Fuller et al reported that food-entrainable circadian rhythms are absent in mice bearing a null mutation of the circadian clock gene Bmal1 and that these rhythms can be rescued by virally-mediated restoration of Bmal1 expression in the dorsomedial nucleus of the hypothalamus (DMH) but not in the suprachiasmatic nucleus (site of the master light-entrainable circadian pacemaker). These results, taken together with controversial DMH lesion results published by the same laboratory, appear to establish the DMH as the site of a Bmal1-dependent circadian mechanism necessary and sufficient for food anticipatory rhythms. However, careful examination of the manuscript reveals numerous weaknesses in the evidence as presented. These problems are grouped as follows and elaborated in detail: 1. data management issues (apparent misalignments of plotted data), 2. failure of evidence to support the major conclusions, and 3. missing data and methodological details. The Fuller et al results are therefore considered inconclusive, and fail to clarify the role of either the DMH or Bmal1 in the expression of food-entrainable circadian rhythms in rodents.

Comment on "Differential Rescue of Light- and Food-Entrainable Circadian Rhythms"

Fuller et al. (Reports, 23 May 2008, p. 1074) reported that the dorsomedial hypothalamus contains a Bmal1-based oscillator that can drive food-entrained circadian rhythms. We report that mice bearing a null mutation of Bmal1 exhibit normal food-anticipatory circadian rhythms. Lack of food anticipation in Bmal1-/- mice reported by Fuller et al. may reflect morbidity due to weight loss, thus raising questions about their conclusions.

Neurobiology of food-entrainable circadian rhythms in mammals

Neurobiology of food-entrainable circadian rhythms in mammals

Two Clocks Are Better Than One

The circadian clock of mammals resides within the hypothalamus, in the suprachiasmatic nucleus, and it is entrained by light. There is evidence that another circadian clock--entrained by food--also exists in the brain, possibly elsewhere in the hypothalamus. By replacing a missing clock component (Bmal) in selected hypothalamic nuclei of mice lacking Bmal1, Fuller et al. identified the dorsomedial nucleus of the hypothalamus as the site of the food-entrained clock. When Bmal1 was reintroduced selectively into the suprachiasmatic nucleus, the previously unresponsive mice regained the ability to entrain both locomotor activity and body temperature rhythms to a 12:12 light:dark cycle. Reintroduction of Bmal into the dorsomedial nucleus restored the ability of the animals to entrain to a restricted period of food availability, but not to a light cycle. Thus, the dorsomedial nucleus contains a second circadian clock that seems to be induced when food is restricted to take over control of functions such as activity levels that increase the chances of the animal successfully locating additional food resources. P. M. Fuller, J. Lu, C. B. Saper, Differential rescue of light- and food-entrainable circadian rhythms. Science 320 , 1074-1077 (2008). [Abstract] [Full Text]

Differential Rescue of Light- and Food-Entrainable Circadian Rhythms

When food is plentiful, circadian rhythms of animals are powerfully entrained by the light-dark cycle. However, if animals have access to food only during their normal sleep cycle, they will shift most of their circadian rhythms to match the food availability. We studied the basis for entrainment of circadian rhythms by food and light in mice with targeted disruption of the clock gene Bmal1, which lack circadian rhythmicity. Injection of a viral vector containing the Bmal1 gene into the suprachiasmatic nuclei of the hypothalamus restored light-entrainable, but not food-entrainable, circadian rhythms. In contrast, restoration of the Bmal1 gene only in the dorsomedial hypothalamic nucleus restored the ability of animals to entrain to food but not to light. These results demonstrate that the dorsomedial hypothalamus contains a Bmal1-based oscillator that can drive food entrainment of circadian rhythms.

Lack of Food Anticipation in Per2 Mutant Mice

Predicting time of food availability is key for survival in most animals. Under restricted feeding conditions, this prediction is manifested in anticipatory bouts of locomotor activity and body temperature. This process seems to be driven by a food-entrainable oscillator independent of the main, light-entrainable clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus . Although the SCN clockwork involves self-sustaining transcriptional and translational feedback loops based on rhythmic expression of mRNA and proteins of clock genes , the molecular mechanisms responsible for food anticipation are not well understood. Period genes Per1 and Per2 are crucial for the SCN's resetting to light . Here, we investigated the role of these genes in circadian anticipatory behavior by studying rest-activity and body-temperature rhythms of Per1 and Per2 mutant mice under restricted feeding conditions. We also monitored expression of clock genes in the SCN and peripheral tissues. Whereas wild-type and Per1 mutant mice expressed regular food-anticipatory activity, Per2 mutant mice did not show food anticipation. In peripheral tissues, however, phase shifts of clock-gene expression in response to timed food restriction were comparable in all genotypes. In conclusion, a mutation in Per2 abolishes anticipation of mealtime, without interfering with peripheral synchronization by feeding cycles.

The dorsomedial hypothalamic nucleus as a putative food-entrainable circadian pacemaker

Temporal restriction of feeding can phase-shift behavioral and physiological circadian rhythms in mammals. These changes in biological rhythms are postulated to be brought about by a food-entrainable oscillator (FEO) that is independent of the suprachiasmatic nucleus. However, the neural substrates of FEO have remained elusive. Here, we carried out an unbiased search for mouse brain region(s) that exhibit a rhythmic expression of the Period genes in a feeding-entrainable manner. We found that the compact part of the dorsomedial hypothalamic nucleus (DMH) demonstrates a robust oscillation of mPer expression only under restricted feeding. The oscillation persisted for at least 2 days even when mice were given no food during the expected feeding period after the establishment of food-entrained behavioral rhythms. Moreover, refeeding after fasting rapidly induced a transient mPer expression in the same area of DMH. Taken in conjunction with recent findings (i) that behavioral expression of food-entrainable circadian rhythms is blocked by cell-specific lesions of DMH in rats and (ii) that DMH neurons directly project to orexin neurons in the lateral hypothalamus, which are essential for proper expression of food-entrained behavioral rhythms, the present study suggests that DMH plays a key role as a central FEO in the feeding-mediated regulation of circadian behaviors.

The dorsomedial hypothalamic nucleus is critical for the expression of food-entrainable circadian rhythms

Circadian rhythms of behavior and physiology can be entrained by daily cycles of restricted food availability, but the pathways that mediate food entrainment are unknown. The dorsomedial hypothalamic nucleus (DMH) is critical for the expression of circadian rhythms and receives input from systems that monitor food availability. Here we report that restricted feeding synchronized the daily rhythm of DMH activity in rats such that c-Fos expression in the DMH was highest at scheduled mealtime. During food restriction, unlesioned rats showed a marked preprandial rise in locomotor activity, body temperature and wakefulness, and these responses were blocked by cell-specific lesions in the DMH. Furthermore, the degree of food entrainment correlated with the number of remaining DMH neurons, and lesions in cell groups surrounding the DMH did not block entrainment by food. These results establish that the neurons of the DMH have a critical role in the expression of food-entrainable circadian rhythms.

Food-entrained circadian rhythms are sustained in arrhythmic Clk/Clk mutant mice.

Daily scheduled feeding is a potent time cue that elicits anticipatory activity in rodents. This food-anticipatory activity (FAA) is controlled by a food-entrainable oscillator (FEO) that is distinct from light-entrained oscillators of the suprachiasmatic nucleus (SCN). Circadian rhythms within the SCN depend on transcription-translation feedback loops in which CLOCK protein is a key positive regulator. The Clock gene is expressed in rhythmic tissues throughout the brain and periphery, implicating its widespread involvement in the functioning of circadian oscillators. To examine whether CLOCK protein is also necessary for the FEO, the effect of daily food restriction was studied in homozygous Clock mutant (Clk/Clk) mice. The results show that Clk/Clk mutant mice exhibit FAA, even when their circadian wheel-running behavior is arrhythmic. As in wild-type controls, FAA in Clk/Clk mutants persists after temporal feeding cues are removed for several cycles, indicating that the FEO is a circadian timer. This is the first demonstration that the Clock gene is not necessary for the expression of a circadian, food-entrained behavior and suggests that the FEO is mediated by a molecular mechanism distinct from that of the SCN.

Food-entrained circadian rhythms in rats are insensitive to deuterium oxide

Rats anticipate a scheduled daily meal by entrainment of a circadian pacemaker separate from the light-entrainable circadian pacemaker located in the suprachiasmatic nuclei (SCN). The site and molecular mechanisms of the food-entrainable pacemaker are unknown. The intrinsic period ( τ) of the SCN pacemaker is significantly lengthened by deuteriation. Sensitivity of food-entrained circadian rhythms to D 2O (25% in drinking water) was evaluated in intact and SCN-ablated rats entrained to daily feeding schedules. In intact rats fed ad-libitum, D 2O lengthened τ sufficiently to drive activity rhythms out of entrainment to the light–dark cycle. By contrast, food-entrained rhythms were surprisingly resistant to modulation by D 2O. The mean daily onset time of food anticipatory activity in rats with complete SCN-ablations was not affected by up to 28 days of D 2O intake. Transient delays and disruption of anticipatory activity were evident in intact and one partial SCN-ablated rat during D 2O treatment, but these are interpretable as effects of coupling and/or masking interactions between a D 2O-sensitive light-entrainable pacemaker, and a D 2O-resistant food-entrained pacemaker. Differential sensitivity to D 2O suggests diversity in the molecular mechanisms of food- and light-entrainable circadian pacemakers in mammals. D 2O may have utility as a screening test to identify putative food-entrainable pacemakers from among those central and peripheral tissues that can express circadian oscillations of clock genes independent of the SCN.

Food-Entrained Feeding and Locomotor Circadian Rhythms in Rats Under Different Lighting Conditions

It has been suggested that two endogenous timekeeping systems, a light-entrainable pacemaker (LEP) and a food-entrainable pacemaker (FEP), control circadian rhythms. To understand the function and interaction between these two mechanisms better, we studied two behavioral circadian rhythmicities, feeding and locomotor activity, in rats exposed to two conflicting zeitgebers, food restriction and light-dark cycles. For this, the food approaches and wheel-running activity of rats kept under light-dark (LD) 12:12, constant darkness (DD), or constant light (LL) conditions and subjected to different scheduled feeding patterns were continuously recorded. To facilitate comparison of the results obtained under the different lighting conditions, the period of the feeding cycles was set in all three cases about Ih less than the light-entrained or free-running circadian rhythms. The results showed that, depending on the lighting conditions, some components of the feeding and wheel-running circadian rhythms could be entrained by food pulses, while others retained their free-running or light-entrained state. Under LD, food pulses had little influence on the light-entrained feeding and loco-motor rhythms. Under DD, relative coordination between free-running and food-associated rhythms may appear. In both cases, the feeding activity associated with the food pulses could be divided into a prominent phase-dependent peak of activity within the period of food availability and another afterward. Wheel-running activity mainly followed the food pulses. Under LL conditions, the food-entrained activity consisted mainly of feeding and wheel-running anticipatory activity. The results provide new evidence that lighting conditions influence the establishment and persistence of food-entrained circadian rhythms in rats. The existence of two coupled pacemakers, LEP and FEP, or a multioscillatory LEP may both explain our experimental results.

Effects of aging on food-entrained circadian rhythms in the rat

Twenty-four hour schedules of restricted food availability entrain a component of the circadian activity rhythm in rats via a food-entrainable pacemaker separate from the light-entrainable pacemaker. The effect of aging on food-entrained circadian rhythms was examined in 6 rats maintained on a restricted diurnal feeding schedule from age 3–21 months and again from 24–25 months. Food-entrainment, measured as behavioral anticipation of a 1-hr daily mealtime during the middle of the light period and persistence of this anticipation rhythm during food deprivation, was apparent in the aged rats when recorded in wheel-running cages from 20–21 months of age. Despite the long duration of restricted diurnal food access, the aged rats, like young rats, rapidly reverted to nocturnal activity when transferred to ad lib feeding. When restricted diurnal feeding was reinstated at 24 months age, these rats, now recorded in food-bin monitoring cages, required more time for a food anticipation pattern to emerge and showed a lower amplitude food anticipation rhythm compared to a group of young adult rats. These age-related changes are similar to those that characterize photically entrained circadian rhythms and suggest that both components of the rat's multioscillatory circadian timekeeping system deteriorate in parallel over the life span.

Characteristics of Food-Entrained Orcadian Rhythms in Rats During Long-Term Exposure to Constant Light

Rats possess a system of circadian oscillators that permit entrainment of circadian activity rhythms independently to 24 hr cycles of light-dark and food access. The nature of interactions between food- and light-entrainable oscillators was examined by observing the generation and persistence of food-entrained circadian rhythms in rats whose light-entrainable rhythms were eliminated by long-term exposure to constant light. Most of these rats showed a delayed generation of food-entrained rhythms and only one of eight animals showed persistence of food associated rhythms during a 4-day food deprivation test. Rats whose light-entrainable rhythms are eliminated by suprachiasmatic nuclei ablation show, in contrast, normal generation and persistence of food-entrained rhythms. The results suggested a disruptive influence of constant light on non-photic entrainment, possibly due to coupling forces between damped light-entrainable oscillators and the food-entrainable oscillators.