Feeding Rhythms Research Articles

The Synergistic Role of Light-Feeding Phase Relations on Entraining Robust Circadian Rhythms in the Periphery.

The feeding and fasting cycles are strong behavioral signals that entrain biological rhythms of the periphery. The feeding rhythms synchronize the activities of the metabolic organs, such as liver, synergistically with the light/dark cycle primarily entraining the suprachiasmatic nucleus. The likely phase misalignment between the feeding rhythms and the light/dark cycles appears to induce circadian disruptions leading to multiple physiological abnormalities motivating the need to investigate the mechanisms behind joint light-feeding circadian entrainment of peripheral tissues. To address this question, we propose a semimechanistic mathematical model describing the circadian dynamics of peripheral clock genes in human hepatocyte under the control of metabolic and light rhythmic signals. The model takes the synergistically acting light/dark cycles and feeding rhythms as inputs and incorporates the activity of sirtuin 1, a cellular energy sensor and a metabolic enzyme activated by nicotinamide adenine dinucleotide. The clock gene dynamics was simulated under various light-feeding phase relations and intensities, to explore the feeding entrainment mechanism as well as the convolution of light and feeding signals in the periphery. Our model predicts that the peripheral clock genes in hepatocyte can be completely entrained to the feeding rhythms, independent of the light/dark cycle. Furthermore, it predicts that light-feeding phase relationship is a critical factor in robust circadian oscillations.

健康と生活習慣病予防における時間栄養学の役割

Almost all metabolic activities of nutrients are under control of the endocrine system, and among them the adrenocortical activity is the most typical example that shows circadian rhythmic changes. We examined relationships between the plasma corticosterone rhythm and feeding schedules in rats. The plasma corticosterone rhythm corresponding to feeding time appears regardless of whether the rats were kept on a light-dark cycle, or in constant light or were blinded. Therefore, it can be concluded that food, more than illumination, is a potent trigger of the circadian adrenocortical rhythm in rats. In addition to the experiments in rats, we also investigated roles of feeding schedulesinformation of the adrenocortical rhythm in human subjects. The corticosterone rhythm disappeared when rats were given a liquid diet intravenously, but not orally, suggesting a necessary role of the oro-gastro-intestinal tract. In addition to the studies in rats, influence to feeding schedules on adrenocortical rhythms was also examined in hospitalized human subjects under a normal light-dark cycle. A clear plasma cortisol rhythm was observed in patients who had been fed on a liquid diet into-duodenum during a restricted time of day, suggest that the circadian cortisol rhythm in man is more closely related to feeding rhythms than cycles of light-dark and sleep-wakefulness. As far as we know, this is the first report demonstrating the important of food in entraining human circadian cortisol rhythm. Next, we measured daily rhythm ofbody temperature in hospitalized patients under continuous and cyclic TEN（totalenteral nutrition）. In the diurnal TEN group, body temperature was low at night and early morning and high in the afternoon early evening. In the continuous TEN group, however, body temperature did not show any significant change with clock-time. In conclusion, the timing of diet intake remarkably modifies the circadian rhythm of body temperature and adrenocortical activity in man. We examined relationship between the feeding time of high salt diet intake and urinary excretion of sodium and chloride. The urinary excretion of sodium and chloride were higher in evening than morning and daytime in woman, is more closely related to the circadian plasma aldosterone rhythm. Furthermore, the clock time difference in endocrine and metabolic responses after physical exercise was assessed according to biorhythm. Physical capacity changed to the time of evaluation, and was greater in the evening than in the morning. Among physiological functions that determine was development of physical strength, plasma growth hormone was higher evening exercise than morning exercise. Further studies are needed on this intriguing idea.

Chronomedicine and type 2 diabetes: shining some light on melatonin.

In mammals, the circadian timing system drives rhythms of physiology and behaviour, including the daily rhythms of feeding and activity. The timing system coordinates temporal variation in the biochemical landscape with changes in nutrient intake in order to optimise energy balance and maintain metabolic homeostasis. Circadian disruption (e.g. as a result of shift work or jet lag) can disturb this continuity and increase the risk of cardiometabolic disease. Obesity and metabolic disease can also disturb the timing and amplitude of the clock in multiple organ systems, further exacerbating disease progression. As our understanding of the synergy between the timing system and metabolism has grown, an interest has emerged in the development of novel clock-targeting pharmaceuticals or nutraceuticals for the treatment of metabolic dysfunction. Recently, the pineal hormone melatonin has received some attention as a potential chronotherapeutic drug for metabolic disease. Melatonin is well known for its sleep-promoting effects and putative activity as a chronobiotic drug, stimulating coordination of biochemical oscillations through targeting the internal timing system. Melatonin affects the insulin secretory activity of the pancreatic beta cell, hepatic glucose metabolism and insulin sensitivity. Individuals with type 2 diabetes mellitus have lower night-time serum melatonin levels and increased risk of comorbid sleep disturbances compared with healthy individuals. Further, reduced melatonin levels, and mutations and/or genetic polymorphisms of the melatonin receptors are associated with an increased risk of developing type 2 diabetes. Herein we review our understanding of molecular clock control of glucose homeostasis, detail the influence of circadian disruption on glucose metabolism in critical peripheral tissues, explore the contribution of melatonin signalling to the aetiology of type 2 diabetes, and discuss the pros and cons of melatonin chronopharmacotherapy in disease management.

Nuclear Proteomics Uncovers Diurnal Regulatory Landscapes in Mouse Liver

SummaryDiurnal oscillations of gene expression controlled by the circadian clock and its connected feeding rhythm enable organisms to coordinate their physiologies with daily environmental cycles. While available techniques yielded crucial insights into regulation at the transcriptional level, much less is known about temporally controlled functions within the nucleus and their regulation at the protein level. Here, we quantified the temporal nuclear accumulation of proteins and phosphoproteins from mouse liver by SILAC proteomics. We identified around 5,000 nuclear proteins, over 500 of which showed a diurnal accumulation. Parallel analysis of the nuclear phosphoproteome enabled the inference of the temporal activity of kinases accounting for rhythmic phosphorylation. Many identified rhythmic proteins were parts of nuclear complexes involved in transcriptional regulation, ribosome biogenesis, DNA repair, and the cell cycle and its potentially associated diurnal rhythm of hepatocyte polyploidy. Taken together, these findings provide unprecedented insights into the diurnal regulatory landscape of the mouse liver nucleus.

Hybrid fish model (Pseudoplatystoma reticulatum♀ × Leiarius marmoratus♂) to study feeding behaviour: protein source self-selection and demand-feeding rhythms

We report feeding studies on hybrid fish (carnivorous × omnivorous) to determine the interactions between protein sources during diet selection trials. We also examined behavioural rhythms (feeding and locomotor activity) using a self-feeding system mounted with a feeder connected to an infrared photocell. Twenty fish (28.20 ± 0.05 g) were distributed in four aquariums (50 L each). First, the system was tested to evaluate the adaptation of catfish (Pseudoplatystoma reticulatum × Leiarius marmoratus) to self-feeding (step I). During this 28-day period, feeding and locomotor activity rhythms were recorded. In the second step, to analyse protein self-selection, a group of 20 fish (19.10 ± 0.05 g) was released in the aquaria. The hybrid catfish were given free access to diets (soya bean meal protein (SP) vs. bovine liver protein (BP)) through two self-feeders. The same procedure was used for (SP) vs. fish meal protein (FP). The catfish learned to activate the feeders to demand a daily feed amount equivalent to 1.8% of their body weight. The fish performed most of their actions (89.0% feeding activity and 94.1 0% locomotor activity) in the night. The diet choice by the catfish became clearer with a significantly lower intake of BP (0.37 g per 100 g body weight, 24.6%) vs. SP (1.06 g per 100 g body weight, 75.4%) and SP (0.61 g per 100 g body weight, 38.9%) vs. FP (0.91 g per 100 g body weight, 61.1%). These findings should be considered when discussing feeding behaviour, nutritional targets and protein sources for future aquaculture feeds.

Daily self-feeding activity rhythms and dietary self-selection of pirarucu (Arapaima gigas)

Daily feeding rhythms and the ability of pirarucu (Arapaima gigas) to compose a balanced diet through macronutrient self-selection were evaluated. Twelve fish (1573.3±74.4g) were distributed in six tanks of 250l, two fish per tank. First, three experimental diets were prepared using an incomplete mixture of macronutrients (75%protein/25%carbohydrate-PC, 75%protein/25%fat-PF and 10% protein/45%fat/45%carbohydrate-PFC). These diets were provided to fish through a self-feeding system connected to a computer to record feeding activity. After this procedure, fish were challenged with a 50% protein dilution (diets: PC50 and PF50). The results showed that pirarucu exhibited a strict diurnal feeding pattern with 95.4% of daily feeding activity observed in the daytime. Fish selected 56.3% P, 24.2% C, 19.5% F, and consumed 150–151kJ/kg BW/day of energy. After protein dilution, fish sustained energy intake by increasing the consumption of PC50 and PF50 to maintain the target protein intake of the previous stage. When fish were protein-restricted, they failed to sustain previous energy intake. These findings can be used to design feeding regimes and for formulating aquafeed for pirarucu. Statement of relevanceThe use of an automatic feeder system activated by the fish could allow the animal to feed at their preferred time and hence reduces waste and improve the intake. Studies that consider the fish preference as a guide in designing diets can provide data on mechanisms of nutrient intake regulation for the development of aquafeeds for new species with potential for aquaculture.

A scalable method for automatically measuring pharyngeal pumping in C. elegans

BackgroundThe nematode Caenorhabditis elegans is widely used for studying small neural circuits underlying behavior. In particular, the rhythmic feeding motions collectively termed pharyngeal pumping are regulated by a nearly autonomous network of 20 neurons of 14 types. Despite much progress achieved through laser ablation, genetics, electrophysiology, and optogenetics, key questions regarding the regulation of pumping remain open. New methodWe describe the implementation and application of a scalable automated method for measuring pumping in controlled environments. Our implementation is affordable and flexible: key hardware and software elements can be modified to accommodate different requirements. ResultsWe demonstrate prolonged measurements under controlled conditions and the resulting high quality data. We show the scalability of our method, enabling high throughput, and its suitability for maintaining static and dynamic conditions. When food availability was oscillated, pumping rates were low as compared to steady conditions and pumping activity was not reliably modulated in response to changes in food concentration. Comparison with existing methodThe prevailing method for measuring rates of pumping relies on scoring by visual inspection of short recordings. Our automated method compares well with manual scoring. It enables detailed statistical characterization under experimental conditions not previously accessible and minimizes unintentional bias. ConclusionsOur approach adds a powerful tool for studying pharyngeal pumping. It enhances the experimental versatility of assaying genetic and pharmacological manipulations and the ability to characterize the resulting behavior. Both the experimental setup and the analysis can be readily adapted to additional challenging motion detection problems.

Late feeding in the active period decreases slow-wave activity

AimsSleep and feeding behaviors closely interact to maintain energy homeostasis. While it is known that sleep disorders can lead to various metabolic issues such as insulin resistance, the mechanism for this effect is poorly understood. We thus investigated whether different feeding rhythms during the active period affect sleep-wake regulation. Main methodsFor 2weeks, mice were randomly assigned to 1 of 3 feeding schedules as follows: free access to lab chow during the active period (ZT12-24, Ad-lib group), free access to lab chow during the first half of the active period (ZT12-18; Morning group), or free access to lab chow during the second half of the active period (ZT18-24, Evening group). Food intake, body weight, body temperature, locomotor activity, and sleep were evaluated. The hypothalamus and cerebral cortex were examined post-mortem. Key findingsNo alterations in food intake or body weight were observed among the 3 groups. The Evening group showed lower slow-wave activity (SWA) than the other 2 groups, in addition to higher expression of orexin mRNA in the hypothalamus and higher concentrations of dopamine and its metabolites in the cerebral cortex. AMPK phosphorylation was increased in the hypothalamus of mice in the Evening group; however, AMPK inhibition had no effect on SWA. SignificanceWe concluded that late feeding reduces SWA in NREM sleep via a mechanism that involves orexin-mediated arousal in the hypothalamus and elevated monoamines in the cerebral cortex. These data have important implications for the relationship between sleep-wake disturbances and metabolic disorders.

Melatonin is a redundant entraining signal in the rat circadian system

The role of melatonin in maintaining proper function of the circadian system has been proposed but very little evidence for such an effect has been provided. To ascertain the role, the aim of the study was to investigate impact of long-term melatonin absence on regulation of circadian system. The parameters of behavior and circadian clocks of rats which were devoid of the melatonin signal due to pinealectomy (PINX) for more than one year were compared with those of intact age-matched controls. PINX led to a decrease in spontaneous locomotor activity and a shortening of the free-running period of the activity rhythm driven by the central clock in the suprachiasmatic nuclei (SCN) in constant darkness. However, the SCN-driven rhythms in activity and feeding were not affected and remained well entrained in the light/dark cycle. In contrast, in these conditions PINX had a significant effect on amplitudes of the clock gene expression rhythms in the duodenum and also partially in the liver. These results demonstrate the significant impact of long-term melatonin absence on period of the central clock in the SCN and the amplitudes of the peripheral clocks in duodenum and liver and suggest that melatonin might be a redundant but effective endocrine signal for these clocks.

Synchronization to light and mealtime of the circadian rhythms of self-feeding behavior and locomotor activity of white shrimps (Litopenaeus vannamei)

The role of light and feeding cycles in synchronizing self-feeding and locomotor activity rhythms was studied in white shrimps using a new self-feeding system activated by photocell trigger. In experiment 1, shrimps maintained under a 12:12h light/dark (LD) photoperiod were allowed to self-feed using feeders connected to a photoelectric cell, while locomotor activity was recorded with a second photocell. On day 30, animals were subjected to constant darkness (DD) for 12days to check the existence of endogenous circadian rhythms. In the experiment 2, shrimps were exposed to both a 12:12h LD photoperiod and a fixed meal schedule in the middle of the dark period (MD, 01:00h). On day 20, shrimps were exposed to DD conditions and the same fixed feeding. On day 30, they were maintained under DD and fasted for 7days. The results revealed that under LD, shrimps showed a clear nocturnal feeding pattern and locomotor activity (81.9% and 67.7% of total daily food-demands and locomotor activity, respectively, at nighttime). Both feeding and locomotor rhythms were endogenously driven and persisted under DD with an average period length (τ) close to 24h (circadian) (τ=24.18±0.13 and 23.87±0.14h for locomotor and feeding, respectively). Moreover, Shrimp showed a daily food intake under LD condition (1.1±0.2gday−1 in the night phase vs. 0.2±0.1gday−1 in the light phase). Our findings might be relevant for some important shrimp aquaculture aspects, such as developing suitable feeding management on shrimp farms.

Effects of chronic forced circadian desynchronization on body weight and metabolism in male mice.

Metabolic functions are synchronized by the circadian clock setting daily patterns of food intake, nutrient delivery, and behavioral activity. Here, we study the impact of chronic jet‐lag (CJL) on metabolism, and test manipulations aimed to overcome potential alterations. We recorded weight gain in C57Bl/6 mice under chronic 6 h advances or delays of the light–dark cycle every 2 days (ChrA and ChrD, respectively). We have previously reported ChrA, but not ChrD, to induce forced desynchronization of locomotor activity rhythms in mice (Casiraghi et al. 2012). Body weight was rapidly increased under ChrA, with animals tripling the mean weight gain observed in controls by day 10, and doubling it by day 30 (6% vs. 2%, and 15% vs. 7%, respectively). Significant increases in retroperitoneal and epidydimal adipose tissue masses (172% and 61%, respectively), adipocytes size (28%), and circulating triglycerides (39%) were also detected. Daily patterns of food and water intake were abolished under ChrA. In contrast, ChrD had no effect on body weight. Wheel‐running, housing of animals in groups, and restriction of food availability to hours of darkness prevented abnormal increase in body weight under ChrA. Our findings suggest that the observed alterations under ChrA may arise either from a direct effect of circadian disruption on metabolism, from desynchronization between feeding and metabolic rhythms, or both. Direction of shifts, timing of feeding episodes, and other reinforcing signals deeply affect the outcome of metabolic function under CJL. Such features should be taken into account in further studies of shift working schedules in humans.

Daily rhythms of digestive enzyme activity and gene expression in gilthead seabream (Sparus aurata) during ontogeny

In order to identify daily changes in digestive physiology in developing gilthead seabream larvae, the enzyme activity (trypsin, lipases and α-amylase) and gene expression (trypsinogen-try, chymotrypsinogen-ctrb, bile salt-activated lipase-cel1b, phospholipase A2-pla2 and α-amylase-amy2a) were measured during a 24h cycle in larvae reared under a 12h light/12h dark photoperiod. Larvae were sampled at 10, 18, 30 and 60days post-hatch. In each sampling day, larvae were sampled every 3h during a complete 24h cycle. The enzyme activity and gene expression exhibited a marked dependent behavior to the light/darkness cycle in all tested ages. The patterns of activity and expression of all tested enzymes were compared to the feeding pattern found in the same larvae, which showed a rhythmic feeding pattern with a strong light synchronization. In the four tested ages, the activities of trypsin, and to a lesser extent lipases and amylase, were related to feeding activity. Molecular expression of the pancreatic enzymes tended to increase during the night, probably as an anticipation of the forthcoming ingestion of food that will take place during the next light period. It follows that the enzymatic activities are being regulated at translational and/or post-translational level. The potential variability of enzyme secretion along the whole day is an important factor to take into account in future studies. A particularly striking consequence of the present results is the reliability of studies based in only one daily sample taken at the same hour of the day, as those focused to assess ontogeny of digestive enzymes.

Self-feeder systems and infrared sensors to evaluate the daily feeding and locomotor rhythms of Pirarucu (Arapaima gigas) cultivated in outdoor tanks

This study evaluated the ability of Pirarucu (Arapaima gigas) to feed through self-demand feeders, and also determined daily feeding rhythm and locomotor activity. Twenty-four fish (312.58±9.55g, mean±SEM) were distributed in six 250-liter outdoor tanks. Each tank was equipped with a feeder adapted to allow fish to self-feed. Self-feeders and occupancy sensors were connected to a computer to allow measurement of feeding and locomotor activity. First, the ability of Pirarucu to use the self-feeding system was assessed. The daily rhythm of feeding activity and locomotion of Pirarucu was then evaluated. The fish learned to trigger the feeders and feeding demand stabilized at 158.79 activations per day. Predominant daytime feeding activity was observed, with 110.86±0.27 counts/day (70% of the total actions) seen during the day, versus 47.93±0.11 counts/day (30%) at night. However, no rhythm in this daily locomotor activity was observed. These study results were based on food being freely available and should be taken into account to improve the supply schedule of the species. Statement of relevanceThe use of an automatic feeder system activated by the fish could allow the animal to feed at their preferred time and hence reduces waste and improve the food intake of Pirarucu raised for aquaculture. Moreover, the optimum feeding strategies, including times, schedules and regimes, should be determined from detailed investigations of feeding rhythms.

Daily feeding and protein metabolism rhythms in Senegalese sole post-larvae.

ABSTRACTFish hatcheries must adapt larval feeding protocols to feeding behavior and metabolism patterns to obtain more efficient feed utilization. Fish larvae exhibit daily ingesting rhythms rather than ingesting food continuously throughout the day. The aim of this study was to determine the daily patterns of feed intake, protein digestibility, protein retention and catabolism in Senegalese sole post-larvae (Solea senegalensis; 33 days post-hatching) using 14C-labeled Artemia protein and incubation in metabolic chambers. Sole post-larvae were fed at 09:00, 15:00, 21:00, 03:00 and 09:00+1 day; and those fed at 09:00, 21:00, 03:00 and 09:00+1 day showed significantly higher feed intake than post-larvae fed at 15:00 h (P=0.000). Digestibility and evacuation rate of ingested protein did not change during the whole cycle (P=0.114); however, post-larvae fed at 21:00 and 03:00 h showed the significantly highest protein retention efficiency and lowest catabolism (P=0.002). Therefore, results confirm the existence of daily rhythmicity in feeding activity and in the utilization of the ingested nutrients in Senegalese sole post-larvae.

Introduction to trophology of walleye pollock

Various aspects of walleye pollock feeding at different stages of its ontogenesis are reviewed and discussed. Structure and functional features of its mouth organs are considered with meristic and quantitative description of the filter one. Lists of pollock prey are presented with attention to changes of food composition and dominant species in the process of its growth from larvae to adult fish. Daily rhythm of feeding is not usual for pollock, but it appears locally and temporary under regional or seasonal conditions of its prey (i.e. plankton). An important feature of the pollock trophology is cannibalism on its own larvae and underyearlings that allows to the population to extend the food spectrum toward small-sized zooplankton. Activity of the pollock feeding can change in several times during the year in dependence on food resources and physiological state of the fish. Year-to-year variability of mass groups of prey (copepods, euphausiids, hyperiids) abundance sometimes coincides with the pollock stock changes in the northern Okhotsk Sea, but variations of arroworms abundance (not important for pollock) never coincide with the stock changes. Daily food ration of pollock (relative to body weight) decreases with its age, but its absolute individual consumption increases that could be a reason of food conversion underestimation. However, large fish are few and feed mostly on fish and benthic invertebrates (similar to cod), so their consumption of zooplankton could be ignored.

Natural selection against a circadian clock gene mutation in mice

Circadian rhythms with an endogenous period close to or equal to the natural light-dark cycle are considered evolutionarily adaptive ("circadian resonance hypothesis"). Despite remarkable insight into the molecular mechanisms driving circadian cycles, this hypothesis has not been tested under natural conditions for any eukaryotic organism. We tested this hypothesis in mice bearing a short-period mutation in the enzyme casein kinase 1ε (tau mutation), which accelerates free-running circadian cycles. We compared daily activity (feeding) rhythms, survivorship, and reproduction in six replicate populations in outdoor experimental enclosures, established with wild-type, heterozygous, and homozygous mice in a Mendelian ratio. In the release cohort, survival was reduced in the homozygote mutant mice, revealing strong selection against short-period genotypes. Over the course of 14 mo, the relative frequency of the tau allele dropped from initial parity to 20%. Adult survival and recruitment of juveniles into the population contributed approximately equally to the selection for wild-type alleles. The expression of activity during daytime varied throughout the experiment and was significantly increased by the tau mutation. The strong selection against the short-period tau allele observed here contrasts with earlier studies showing absence of selection against a Period 2 (Per2) mutation, which disrupts internal clock function, but does not change period length. These findings are consistent with, and predicted by the theory that resonance of the circadian system plays an important role in individual fitness.

The role of feeding rhythm, adrenal hormones and neuronal inputs in synchronizing daily clock gene rhythms in the liver

The master clock in the hypothalamic suprachiasmatic nucleus (SCN) is assumed to distribute rhythmic information to the periphery via neural, humoral and/or behavioral connections. Until now, feeding, corticosterone and neural inputs are considered important signals for synchronizing daily rhythms in the liver. In this study, we investigated the necessity of neural inputs as well as of the feeding and adrenal hormone rhythms for maintaining daily hepatic clock gene rhythms. Clock genes kept their daily rhythm when only one of these three signals was disrupted, or when we disrupted hepatic neuronal inputs together with the adrenal hormone rhythm or with the daily feeding rhythm. However, all clock genes studied lost their daily expression rhythm after simultaneous disruption of the feeding and adrenal hormone rhythm. These data indicate that either a daily rhythm of feeding or adrenal hormones should be present to synchronize clock gene rhythms in the liver with the SCN.

Controlling the Cyanobacterial Clock by Synthetically Rewiring Metabolism.

Circadian clocks are oscillatory systems that allow organisms to anticipate rhythmic changes in the environment. Several studies have shown that circadian clocks are connected to metabolism, but it is not generally clear whether metabolic signaling is one voice among many that influence the clock or whether metabolic cycling is the major clock synchronizer. To address this question in cyanobacteria, we used a synthetic biology approach to make normally autotrophic cells capable of growth on exogenous sugar. This allowed us to manipulate metabolism independently from light and dark. We found that feeding sugar to cultures blocked the clock-resetting effect of a dark pulse. Furthermore, in the absence of light, the clock efficiently synchronizes to metabolic cycles driven by rhythmic feeding. We conclude that metabolic activity, independent of its source, is the primary clock driver in cyanobacteria.

Effects of 6-meals-a-day feeding and 6-meals-a-day feeding combined with adrenalectomy on daily gene expression rhythms in rat epididymal white adipose tissue.

The master clock in the hypothalamic suprachiasmatic nucleus (SCN) is assumed to synchronize the tissue-specific rhythms of the peripheral clocks with the environmental day/night changes via neural, humoral and/or behavioral connections. The feeding rhythm is considered an important Zeitgeber for peripheral clocks, as daytime feeding reverses (clock) gene rhythms in the periphery, but not in the SCN. In this study, we investigated the necessity of a daily feeding rhythm for maintaining gene expression rhythms in epididymal white adipose tissue (eWAT). We showed that 7 of 9 rhythmic metabolic/adipokine genes, but not clock genes, lost their rhythmicity upon exposure to 6-meals-a-day feeding. Previously, we showed comparable effects of adrenalectomy on eWAT; therefore, subsequently we investigated the effect of simultaneous disruption of these humoral and behavioral signaling pathways, by exposing adrenalectomized animals to 6-meals-a-day feeding. Interestingly, under these conditions, all the clock genes and 10 of 11 rhythmic metabolic/adipokine genes lost their rhythmicity. These data indicate that adrenal hormones and feeding rhythm are indispensable for maintaining daily rhythms in metabolic/adipokine gene, but not clock gene, expression in eWAT. In contrast, at least one of these two signals should be present in order for eWAT clock gene rhythms to be maintained.

Circadian and feeding rhythms differentially affect rhythmic mRNA transcription and translation in mouse liver

Diurnal oscillations of gene expression are a hallmark of rhythmic physiology across most living organisms. Such oscillations are controlled by the interplay between the circadian clock and feeding rhythms. Although rhythmic mRNA accumulation has been extensively studied, comparatively less is known about their transcription and translation. Here, we quantified simultaneously temporal transcription, accumulation, and translation of mouse liver mRNAs under physiological light-dark conditions and ad libitum or night-restricted feeding in WT and brain and muscle Arnt-like 1 (Bmal1)-deficient animals. We found that rhythmic transcription predominantly drives rhythmic mRNA accumulation and translation for a majority of genes. Comparison of wild-type and Bmal1 KO mice shows that circadian clock and feeding rhythms have broad impact on rhythmic gene expression, Bmal1 deletion affecting surprisingly both transcriptional and posttranscriptional levels. Translation efficiency is differentially regulated during the diurnal cycle for genes with 5'-Terminal Oligo Pyrimidine tract (5'-TOP) sequences and for genes involved in mitochondrial activity, many harboring a Translation Initiator of Short 5'-UTR (TISU) motif. The increased translation efficiency of 5'-TOP and TISU genes is mainly driven by feeding rhythms but Bmal1 deletion also affects amplitude and phase of translation, including TISU genes. Together this study emphasizes the complex interconnections between circadian and feeding rhythms at several steps ultimately determining rhythmic gene expression and translation.