Food-entrainable Oscillator Research Articles

Isolating Neural Correlates of the Pacemaker for Food Anticipation

Mice fed a single daily meal at intervals within the circadian range exhibit food anticipatory activity. Previous investigations strongly suggest that this behaviour is regulated by a circadian pacemaker entrained to the timing of fasting/refeeding. The neural correlate(s) of this pacemaker, the food entrainable oscillator (FEO), whether found in a neural network or a single locus, remain unknown. This study used a canonical property of circadian pacemakers, the ability to continue oscillating after removal of the entraining stimulus, to isolate activation within the neural correlates of food entrainable oscillator from all other mechanisms driving food anticipatory activity. It was hypothesized that continued anticipatory activation of central nuclei, after restricted feeding and a return to ad libitum feeding, would elucidate a neural representation of the signaling circuits responsible for the timekeeping component of the food entrainable oscillator. Animals were entrained to a temporally constrained meal then placed back on ad libitum feeding for several days until food anticipatory activity was abolished. Activation of nuclei throughout the brain was quantified using stereological analysis of c-FOS expressing cells and compared against both ad libitum fed and food entrained controls. Several hypothalamic and brainstem nuclei remained activated at the previous time of food anticipation, implicating them in the timekeeping mechanism necessary to track previous meal presentation. This study also provides a proof of concept for an experimental paradigm useful to further investigate the anatomical and molecular substrates of the FEO.

Synchronization of PER1 protein in parabrachial nucleus in a natural model of food anticipatory activity

Rabbit pups represent a natural model of food anticipatory activity (FAA). FAA is the behavioral output of a putative food entrainable oscillator (FEO). It had been suggested that the FEO is comprised of a distributed system of clocks that work in concert in response to gastrointestinal input by food. Scheduled food intake synchronizes several nuclei in the brain, and the hypothalamus has received particular attention. On the contrary, brainstem nuclei, despite being among the brain structures to first receive food cues, have been scarcely studied. Here we analysed by immunohistochemistry possible oscillation of FOS and PER1 proteins through a complete 24-h cycle in the dorsal vagal complex (DVC) and parabrachial nucleus (PBN) of 7-8-day-old rabbit pups scheduled to nurse during the night (02:00 h) or day (10:00 h), and also in fasted subjects to explore the possible persistence of oscillations. We found a clear induction of FOS that peaks 1.5 h after nursing in all nuclei studied. PER1 was only synchronized in the PBN, reaching highest values 12 h after nursing. Only PER1 oscillations persisted, with a shift, in fasted subjects. We conclude that the DVC nuclei are probably more related to the transmission of food cues to other brain regions, but that the PBN participates in the integration of information essential for FAA. Our results support previous findings suggesting that the DVC nuclei, but not PBN, are not essential for FAA. We suggest that PBN is a key component of the proposed distributed system of clocks involved in FAA.

Daytime restricted feeding modifies 24 h rhythmicity and subcellular distribution of liver glucocorticoid receptor and the urea cycle in rat liver

The timing system in mammals is formed by a set of peripheral biological clocks coordinated by a light-entrainable pacemaker located in the suprachiasmatic nucleus. Daytime restricted feeding (DRF) modifies the circadian control and uncouples the light-dependent physiological rhythmicity, food access becoming the principal external time cue. In these conditions, an alternative biological clock is expressed, the food-entrainable oscillator (FEO). Glucocorticoid hormones are an important part of the humoral mechanisms in the daily synchronisation of the metabolic response of peripheral oscillators by the timing system. A peak of circulating corticosterone has been reported before food access in DRF protocols. In the present study we explored in the liver the 24h variations of: (1) the subcellular distribution of glucocorticoid receptor (GCR), (2) the activities of the corticosterone-forming and NADPH-generating enzymes (11β-hydroxysteroid dehydrogenase type 1 (11β-HSD-1) and hexose-6-phosphate dehydrogenase (H6PDH)), and, (3) parameters related with the urea cycle (circulating urea and activities of carbamoyl phosphate synthetase and ornithine transcarbamylase) elicited by DRF. The results showed that DRF promoted an increase of more than two times of the hepatic GCR, but exclusively in the cytosolic compartment, since the GCR in the nuclear fraction showed a reduction. No changes were observed in the activities of 11β-HSD-1 and H6PDH, but the rhythmicity of all of the urea cycle-related parameters was modified. It is concluded that liver glucocorticoid signalling and the urea cycle are responsive to feeding-restricted schedules and could be part of the FEO.

Restricted food access during the daytime modifies the 24-h rhythmicity of apoptosis and cellular duplication in rat liver

Twenty-four hour rhythmicity of liver metabolic and cellular activities is greatly modified by daytime restricted feeding (DRF). The use of this protocol has been associated with the study and characterization of the food-entrained oscillator (FEO). The aim of this project was to explore the adaptations in hepatic apoptotic activity and cellular duplication in a 2-h restricted-feeding schedule. Our findings indicate that DRF promoted a rise in the intrinsic pathway of apoptosis which is dependent of mitochondrial cytochrome C release. The group synchronized by food (FS) also showed an increased activity of caspase-3, but at the same time an augmented number of liver cells positive to cellular duplication marker (PCNA), proliferative cell nuclear antigen. The data strongly suggest that DRF/FEO expression causes an enhancement of cellular exchange in the liver.

Food entrainment: major and recent findings

Mammals exhibit daily anticipatory activity to cycles of food availability. Studies on such food anticipatory activity (FAA) have been conducted mainly in nocturnal rodents. They have identified FAA as the behavioral output of a food entrained oscillator (FEO), separate of the known light entrained oscillator (LEO) located in the suprachiasmatic nucleus (SCN) of hypothalamus. Here we briefly review the main characteristics of FAA. Also, we present results on four topics of food anticipation: (1) possible input signals to FEO, (2) FEO substrate, (3) the importance of canonical clock genes for FAA, and (4) potential practical applications of scheduled feeding. This mini review is intended to introduce the subject of food entrainment to those unfamiliar with it but also present them with relevant new findings on the issue.

Bmal1in the Nervous System Is Essential for Normal Adaptation of Circadian Locomotor Activity and Food Intake to Periodic Feeding

Temporal restriction of feeding can entrain circadian behavioral and physiological rhythms in mammals. These changes in biological rhythms are postulated to be brought about by a putative food-entrainable oscillator (FEO) that is independent of the suprachiasmatic nucleus (SCN). However, the anatomical substrates and molecular machinery of FEO remain elusive. We report here that mice with a nervous system-specific deletion of Bmal1, an essential clock component, had a marked deficit in entrainment of locomotor activity by periodic feeding, accompanied by reduced food intake and subsequent loss of body weight. These mice exhibited a nearly normal light-entrainable activity rhythm driven by the SCN, because deletion of the Bmal1 gene in the SCN was only partial. These findings suggest that an SCN-independent FEO in the nervous system requires Bmal1 and plays a critical role in adaptation of circadian locomotor activity and food intake to periodic feeding.

The firing rate of hippocampal CA1 place cells is modulated with a circadian period

The accurate recall of an event is usually dependent on a memory trace that encodes three pieces of information; what happened, when the event happened, and where. The established phenomenology of hippocampal CA1 pyramidal neurons could reflect mechanisms via which some of this information (where and what) is encoded; but so far there has been little evidence for a mechanism by which these cells might represent "when." It was therefore of interest to examine the activity of CA1 neurons over a substantial temporal duration. Forty-eight CA1 neurons were recorded once an hour during long (24-48 h) exposures to a single, stable environment where minimal time-of-day cues were available. Only data from the first 25 h of recording was analyzed quantitatively. We found that the mean ensemble firing rate of these cells changed predictably such that it was closely correlated (r = 0.707) to a reference sine wave with a 25-h period and a positive peak at recording start. This relationship was not explained by changes in the animal's running speed or amount of the recording environment covered in each recording session. When data were referenced to the onset or offset of the normal light-on period, the correlation with the sinusoid was abolished. At an individual cell level, the majority of neurons (n = 31) had significant correlations (P < 0.05) with the reference sine. We conclude that the firing rate of a large proportion of cells in area CA1 of the hippocampus are modulated over a circadian period but that this modulation is not entrained to light. Rather, entry into the environment and the associated food availability appear to be the entraining factors. We hypothesize that these neurons may be part of the putative food-entrainable oscillator. Such a system could enable an animal to discriminate between spatial representations on a temporal dimension with reference to the time of food availability.

Time-imposed daily restricted feeding induces rhythmic expression of Fgf21 in white adipose tissue of mice

Fibroblast growth factor 21 (FGF21) is a key metabolic regulator that is induced by fasting and starvation, and its expression is thought to be regulated by the circadian clock in the liver. To evaluate the functional role of FGF21 in the circadian regulation of physiology and behavior, we examined the temporal expression profiles of Fgf21 and circadian clock genes in addition to behavioral activity rhythms under adlibitum feeding (ALF) and time-imposed restricted feeding (RF) in mice. Four hours of daily restricted feeding during the daytime induced over an 80-fold increase in feeding-dependent rhythmic Fgf21 mRNA expression in epididymal white adipose tissue (eWAT), although the expression levels were continuously increased 10-fold in the liver of wild-type (WT) mice. Refeeding subsequent to transient fasting revealed that refeeding but not fasting remarkably induces Fgf21 expression in eWAT, although fasting-induced hepatic Fgf21 expression is completely reversed by refeeding. The free-running period of locomotor activity rhythm under ALF and the food anticipatory activity (FAA) under RF remained intact in Fgf21 knockout (KO) mice, suggesting that FGF21 is dispensable for both the central clock in the suprachiasmatic nucleus (SCN) and the food-entrainable oscillator that governs the FAA. Temporal expression profiles of circadian genes such as mPer2 and BMAL1 were essentially identical in both tissues between WT and Fgf21 KO mice under RF. The physiological role of the refeeding-induced adipose Fgf21 expression remains to be elucidated.

Endocannabinoids Gate State-Dependent Plasticity of Synaptic Inhibition in Feeding Circuits

Changes in food availability alter the output of hypothalamic nuclei that underlie energy homeostasis. Here, we asked whether food deprivation impacts the ability of GABA synapses in the dorsomedial hypothalamus (DMH), an important integrator of satiety signals, to undergo activity-dependent changes. GABA synapses in DMH slices from satiated rats exhibit endocannabinoid-mediated long-term depression (LTD(GABA)) in response to high-frequency stimulation of afferents. When CB1Rs are blocked, however, the same stimulation elicits long-term potentiation (LTP(GABA)), which manifests presynaptically and requires heterosynaptic recruitment of NMDARs and nitric oxide (NO). Interestingly, NO signaling is required for eCB-mediated LTD(GABA). Twenty-four hour food deprivation results in a CORT-mediated loss of CB1R signaling and, consequently, GABA synapses only exhibit LTP(GABA). These observations indicate that CB1R signaling promotes LTD(GABA) and gates LTP(GABA). Furthermore, the satiety state of an animal, through regulation of eCB signaling, determines the polarity of activity-dependent plasticity at GABA synapses in the DMH.

Sucrose modifies c-fos mRNA expression in the brain of rats maintained on feeding schedules

Food intake is regulated according to circadian activity, metabolic needs and the hedonic value of food. Rodents placed on a fixed feeding schedule show behavioral and physiological anticipation of mealtime referred to as food-anticipatory activity (FAA). FAA is driven by the food-entrainable oscillator (FEO), whose anatomical substrate is not yet known. Recent data have shown that restricted feeding schedules for regular chow and daily limited access to palatable food in free-feeding rats activate distinct brain regions during FAA. The combination of a deprivation regimen and scheduled access to palatable food may give rise to a more global anticipatory mechanism because the temporal cycles of energy balance would be strongly modulated by the incentive properties of palatable food; however, the neuronal response to this combined treatment is not yet known. The present study investigated how adding palatable sucrose to feeding schedules affects the pattern of brain c-fos mRNA expression during FAA (0-3 h) and 1 h following feeding. The rats maintained on scheduled chow access increased their daily chow intake, while the rats maintained on scheduled sucrose and chow mainly increased their daily sucrose intake. Adding sucrose to scheduled feeding displaced c-fos mRNA expression from the dorsomedial and paraventricular hypothalamic nuclei and posterior lateral hypothalamus (LH) to the prefrontal cortex, lateral septum, nucleus accumbens and anterior LH. During refeeding, the rats on scheduled sucrose demonstrated higher activation of the nucleus of the solitary tract. The present results suggest that palatable sucrose combined with restricted feeding schedules activate a distinct neuronal network compared to neuronal activation produced by scheduled access to regular chow. These data provide evidence that the brain may contain different food-oscillatory systems and that food palatability may shift the neuronal activity from the medial hypothalamus to the limbic and reward-related areas even at the negative metabolic state.

Food anticipation depends on oscillators and memories in both body and brain

Despite the importance of learning and circadian rhythms to feeding, there has been relatively little effort to integrate these separate lines of research. In this review, we focus on how light and food entrainable oscillators contribute to the anticipation of food. In particular, we examine the evidence for temporal conditioning of food entrainable oscillators throughout the body. The evidence suggests a shift away from previous notions of a single locus or neural network of food entrainable oscillators to a distributed system involving dynamic feedback among cells of the body and brain. Several recent advances, including documentation of peroxiredoxin metabolic circadian oscillation and anticipatory behavior in the absence of a central nervous system, support the possibility of conditioned signals from the periphery in determining anticipatory behavior. Individuals learn to detect changes in internal and external signals that occur as a consequence of the brain and body preparing for an impending meal. Cues temporally near and far from actual energy content can then be used to optimize responses to temporally predictable and unpredictable cues in the environment.

Interaction between hypothalamic dorsomedial nucleus and the suprachiasmatic nucleus determines intensity of food anticipatory behavior

Food anticipatory behavior (FAA) is induced by limiting access to food for a few hours daily. Animals anticipate this scheduled meal event even without the suprachiasmatic nucleus (SCN), the biological clock. Consequently, a food-entrained oscillator has been proposed to be responsible for meal time estimation. Recent studies suggested the dorsomedial hypothalamus (DMH) as the site for this food-entrained oscillator, which has led to considerable controversy in the literature. Herein we demonstrate by means of c-Fos immunohistochemistry that the neuronal activity of the suprachiasmatic nucleus (SCN), which signals the rest phase in nocturnal animals, is reduced when animals anticipate the scheduled food and, simultaneously, neuronal activity within the DMH increases. Using retrograde tracing and confocal analysis, we show that inhibition of SCN neuronal activity is the consequence of activation of GABA-containing neurons in the DMH that project to the SCN. Next, we show that DMH lesions result in a loss or diminution of FAA, simultaneous with increased activity in the SCN. A subsequent lesion of the SCN restored FAA. We conclude that in intact animals, FAA may only occur when the DMH inhibits the activity of the SCN, thus permitting locomotor activity. As a result, FAA originates from a neuronal network comprising an interaction between the DMH and SCN. Moreover, this study shows that the DMH-SCN interaction may serve as an intrahypothalamic system to gate activity instead of rest overriding circadian predetermined temporal patterns.

Attenuated Food Anticipatory Activity and Abnormal Circadian Locomotor Rhythms in Rgs16 Knockdown Mice

Regulators of G protein signaling (RGS) are a multi-functional protein family, which functions in part as GTPase-activating proteins (GAPs) of G protein α-subunits to terminate G protein signaling. Previous studies have demonstrated that the Rgs16 transcripts exhibit robust circadian rhythms both in the suprachiasmatic nucleus (SCN), the master circadian light-entrainable oscillator (LEO) of the hypothalamus, and in the liver. To investigate the role of RGS16 in the circadian clock in vivo, we generated two independent transgenic mouse lines using lentiviral vectors expressing short hairpin RNA (shRNA) targeting the Rgs16 mRNA. The knockdown mice demonstrated significantly shorter free-running period of locomotor activity rhythms and reduced total activity as compared to the wild-type siblings. In addition, when feeding was restricted during the daytime, food-entrainable oscillator (FEO)-driven elevated food-anticipatory activity (FAA) observed prior to the scheduled feeding time was significantly attenuated in the knockdown mice. Whereas the restricted feeding phase-advanced the rhythmic expression of the Per2 clock gene in liver and thalamus in the wild-type animals, the above phase shift was not observed in the knockdown mice. This is the first in vivo demonstration that a common regulator of G protein signaling is involved in the two separate, but interactive circadian timing systems, LEO and FEO. The present study also suggests that liver and/or thalamus regulate the food-entrained circadian behavior through G protein-mediated signal transduction pathway(s).

Feeding Time Synchronizes Clock Gene Rhythmic Expression in Brain and Liver of Goldfish (Carassius auratus)

Little is known about the feeding time dependence of clock gene expression in fish. The aim of the present study was to investigate whether a scheduled feeding time can entrain the rhythmic expression of several clock genes (period and cryptocrome) in the brain and liver of a teleost, the goldfish. Fish maintained under continuous light (LL) conditions were divided into 3 groups. Two groups were fed daily at 1000 h and 2200 h, respectively, and the third group was subjected to a random schedule regime. After 30 days, the fishes under 24-h food deprivation were sacrificed through a 24-h cycle, and clock gene expression in the optic tectum, hypothalamus, and liver was quantified by real-time PCR. The findings pointed to differences between the central and peripheral tissues studied. In the absence of a light-dark cycle (constant light), a scheduled feeding regime was necessary and sufficient to maintain both the rhythmic expression of several clock genes in the optic tectum and hypothalamus, as well as daily rhythms in locomotor activity. In contrast, neither locomotor activity nor clock gene expression in brain tissues was synchronized in randomly fed fish. However, in the liver, most of the clock genes studied presented significant daily rhythms in phase (related to the time of the last meal) in all 3 experimental groups, suggesting that the daily rhythm of clock genes in this organ only depends on the last meal time. The data suggest that, as in mammals, the smooth running of the food entrainable oscillator (FEO) in fish involves the rhythmic expression of several clock genes (Per1 and Cry3) in the central and peripheral structures. The results also indicate that the food anticipatory activity (FAA) in goldfish is not only the result of rhythmic clock gene expression in the liver because rhythmic clock gene expression was observed in randomly fed fishes, while FAA was not observed.

Changes in the 24 h Rhythmicity of Liver PPARs and Peroxisomal Markers When Feeding Is Restricted to Two Daytime Hours

Restricted feeding (RF) during daytime is associated with anticipatory activity before feeding, marked hyperphagia after mealtime, adjustments in hepatic metabolism, and the expression of a food-entrained oscillator (FEO). 24 h rhythmicity of liver PPARα, β, and γ, peroxisomal markers (PMP70, AOX, and catalase), and free fatty acids (FFAs) during RF was evaluated. The effect of fasting-refeeding was also studied. Results showed (1) higher levels of FFA before feeding, (2) a shift of PPARα and PPARγ before and of PPARβ peaks after feeding, (3) an increase in peroxisomal markers, (4) a shift of PMP70 and AOX peaks before feeding, and of maximal catalase activity in the dark period, (5) changes in the fasting-refeeding response, and (6) high correlations (>0.9) of serum corticosterone with PPARα and PPARγ and of PMP70 with PPARβ. In conclusion, 24 h rhythmicity of FFA, liver PPARs, and peroxisomal markers are biochemical adaptations associated with daytime RF and FEO expression.

Circadian Organization Is Governed by Extra-SCN Pacemakers

In mammals, a pacemaker in the suprachiasmatic nucleus (SCN) is thought to be required for behavioral, physiological, and molecular circadian rhythms. However, there is considerable evidence that temporal food restriction (restricted feedisng [RF]) and chronic methamphetamine (MA) can drive circadian rhythms of locomotor activity, body temperature, and endocrine function in the absence of SCN. This indicates the existence of extra-SCN pacemakers: the Food Entrainable Oscillator (FEO) and Methamphetamine Sensitive Circadian Oscillator (MASCO). Here, we show that these extra-SCN pacemakers control the phases of peripheral oscillators in intact as well as in SCN-ablated PER2::LUC mice. MA administration shifted the phases of SCN, cornea, pineal, pituitary, kidney, and salivary glands in intact animals. When the SCN was ablated, disrupted phase relationships among peripheral oscillators were reinstated by MA treatment. When intact animals were subjected to restricted feeding, the phases of cornea, pineal, kidney, salivary gland, lung, and liver were shifted. In SCN-lesioned restricted-fed mice, phases of all of the tissues shifted such that they aligned with the time of the meal. Taken together, these data show that FEO and MASCO are strong circadian pacemakers able to regulate the phases of peripheral oscillators.

Food- and light-entrainable oscillators control feeding and locomotor activity rhythms, respectively, in the Japanese catfish, Plotosus japonicus

Feeding and locomotor activities of the Japanese catfish Plotosus japonicus under solitary condition were recorded to identify mechanisms controlling these behaviours. In the absence of food, the catfish showed nocturnal locomotor activity, but no feeding activity. Under ad libitum food conditions, both feeding and locomotor activities occurred during the dark period and were synchronized with light/dark (LD) cycles. Feeding activity lasted for 11-24 days when food was stopped after ad libitum food availability. Restricted food during the light phase produced both food-anticipatory and light-entrainable feeding activity. Furthermore, this condition produced weak food-anticipatory and light-entrainable locomotor activity. Under the light/light (LL) condition, restricted food produced food-anticipatory feeding and locomotor activities, suggesting that a food-entrainable oscillator controls both feeding and locomotor activities. However, under the LL condition, light-entrainable feeding and locomotor activities were not observed, suggesting that a light-entrainable oscillator controls both feeding and locomotor activities. During a restricted food schedule, LD cycle shifts resulted in disrupted synchronization of feeding activity onset in three of the four fish, but one fish showed synchronized feeding activity. These results suggest that the food- and the light-entrainable oscillator may control feeding and locomotor activities, respectively.

Disruption of Peripheral Circadian Timekeeping in a Mouse Model of Huntington's Disease and Its Restoration by Temporally Scheduled Feeding

Behavioral circadian rhythms disintegrate progressively in the R6/2 mouse model of Huntington's disease (HD), recapitulating the sleep-wake disturbance seen in HD patients. Here we show that disturbances in circadian pacemaking are not restricted to the brain, but also encompass peripheral metabolic pathways in R6/2 mice. Notably, circadian rhythms of clock-driven genes that are key metabolic outputs in the liver are abolished in vivo. This deficiency is accompanied by arrhythmic expression of the clock genes Cry1 and Dbp, and a phase-advanced Per2 cycle. Compromised circadian metabolic cycles are not, however, a consequence of deficient pacemaking intrinsic to the liver, because when cultured in vitro, R6/2 liver slices exhibit self-sustained circadian bioluminescence rhythms. We therefore propose that compromised metabolic cycles arise from an internal desynchronization secondary to altered feeding patterns and impaired circadian signaling from the central pacemaker of the suprachiasmatic nucleus (SCN). Importantly, the SCN-independent food-entrainable oscillator remains intact in R6/2 mice and, when invoked, can restore daily behavioral cycles and reverse some of the metabolic abnormalities seen in the liver. Disturbances of metabolism have long been thought to be an important feature of HD. Uncoupling liver metabolism from circadian drives will reduce metabolic efficiency and cause imbalances in metabolites known to be deleterious to brain function. Thus, even subtle imbalances in liver function may exacerbate symptoms of HD, where neurological function is already compromised.

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.

Behavioural food anticipation in clock genes deficient mice: confirming old phenotypes, describing new phenotypes

Animals fed daily at the same time exhibit circadian food-anticipatory activity (FAA), which has been suggested to be driven by one or several food-entrainable oscillators (FEOs). FAA is altered in mice lacking some circadian genes essential for timekeeping in the main suprachiasmatic clock (SCN). Here, we confirmed that single mutations of clock genes Per1(-/-) and Per2(Brdm1) alter FAA expression in constant darkness (DD) or under a light-dark cycle (LD). Furthermore, we found that Per1(-/-);Per2(Brdm1) and Per2(Brdm1);Cry1(-/-) double mutant animals did not display a stable and significant FAA either in DD or LD. Interestingly, rescued behavioural rhythms in Per2(Brdm1);Cry2(-/-) mice in DD were totally entrained to feeding time and re-synchronized after phase-shifts of mealtime, indicating a higher SCN sensitivity to feeding cues. However, under an LD cycle and restricted feeding at midday, FAA in double Per2(Brdm1);Cry2(-/-) mutant mice was absent. These results indicate that shutting down one or two clock genes results in altered circadian meal anticipation. Moreover, we show that in a genetically rescued SCN clock (Per2(Brdm1);Cry2(-/-)), food is a powerful zeitgeber to entrain behavioural rhythms, leading the SCN to be more sensitive to feeding cues than in wild-type littermates.