Growth, body composition and hepatic tyrosine aminotransferase activity in cortisol-fed channel catfish, Ictalurus punctatus Rafinesque

Metabolic consequences of chronic elevation of cortisol in the diet of yearling channel catfish, Ictalurus punctatus , were studied. Cortisol was incorporated into the diet in concentrations of 1, 10, 50 and 100 μg/g of food. This diet was offered at 3% of the body weight per day for 10 weeks. Fish were individually weighed and measured at 2-week intervals and feeding rates were adjusted. Body weight, liposomatic index and condition factor were significantly lower and food conversion was significantly higher in fish fed 50 or 100 μng cortisol/g of food when compared with controls. The hepatosomatic index of fish fed cortisol at the rate of 100 μg/g of food decreased significantly. Specific activity of hepatic tyrosine aminotransferase was significantly higher at the two highest cortisol doses. Long-term cortisol administration can reduce growth and condition factor by activating gluconeogenic mechanisms in which lipids and amino acids, rather than carbohydrates, are used for energy production. The metabolic effects of exogenous cortisol in this study offer an explanation for the decreased growth of fish under conditions that activate the secretion of endogenous cortisol.     

Effects of cortisol on aggression and locomotor activity in rainbow trout

Noninvasive administration of cortisol through the diet resulted in relatively rapid (<1.5 h) and highly reproducible increases in plasma cortisol in rainbow trout, comparable to changes seen in fish subjected to substantial stress. Juvenile rainbow trout were reared in isolation for 1 week, before their daily food ration was replaced by a meal of cortisol-treated food corresponding to 6 mg cortisol kg(-1). All fish were observed for 30 min, beginning at 1 or 48 h following the introduction of cortisol-treated food. Additional cortisol (75% of the original dose on Day 2, and 50% on Day 3) was administered to the long-term cortisol-treated group. The resulting blood plasma concentrations of cortisol were similar in short- and long-term treated fish, and corresponded to those previously seen in stressed rainbow trout. Controls were fed similar food without cortisol. Half of the fish from each treatment group (controls and short- and long-term cortisol) were subjected to an intruder test (a smaller conspecific introduced into the aquarium), while half of the fish were observed in isolation. In fish challenged by a conspecific intruder, short-term cortisol treatment stimulated locomotor activity, while long-term treatment inhibited locomotion. Aggressive behavior was also inhibited by long-term cortisol treatment, but not by short-term exposure to cortisol. Cortisol treatment had no effect on locomotor activity in undisturbed fish, indicating that the behavioral effects of cortisol were mediated through interaction with other signal systems activated during the simulated territorial intrusion test. This study demonstrates for the first time that cortisol has time- and context-dependent effects on behavior in teleost fish.     

Regulation of protogynous sex change by competition between corticosteroids and androgens: an experimental test using sandperch, Parapercis cylindrica

Cortisol, the dominant corticosteroid in fish, and 11-ketotestosterone (11KT), the most potent androgen in fish, are both synthesized and (or) deactivated by the same two enzymes, 11beta-hydroxylase and 11beta-hydroxysteroid dehydrogenase. Cortisol is synthesized in response to stress (such as that caused by interaction with a dominant conspecific), whereas 11KT is synthesized during protogynous sex change. It has been hypothesized that corticosteroids (such as cortisol) inhibit 11KT synthesis via substrate competition, thereby providing a mechanism for the regulation of socially mediated, protogynous sex change. We tested this hypothesis by administering cortisol (50 microg g(-1) body weight) to female sandperch (Parapercis cylindrica) under social conditions that were permissive to sex change (i.e. in the absence of suppressive male dominance). Twenty-one days later, mean physiological cortisol concentration in cortisol-treated fish was 4.2-fold greater than that in 'socially stressed' female fish maintained in a semi-natural system. Although the dosage of cortisol was therefore considered to be favorable for engendering competitive inhibition of 11KT synthesis, all cortisol-treated fish changed sex, as did all sham-treated and control fish (n=7 fish per treatment). In addition, there was no effect of cortisol treatment on the rate of sex change or on the pattern of steroidogenesis. Thus, our results refute the hypothesis that protogynous sex change is regulated by substrate competition between corticosteroids and androgens.     

The effects of feeding regime on proximate body composition and patterns of energy deposition in juvenile Arctic charr, Salvelinus alpinus

Juvenile Arctic charr, Salvelinus alpinus , subjected to either food deprivation or restricted feeding showed reductions in relative sizes of both liver and viscera. With food deprivation, percentage lipid decreased and percentage water increased in the eviscerated carcass, which is in agreement with findings for several other fish species. Following transfer from a restricted to a satiation feeding regime, the relative sizes of both liver and viscera increased to levels exceeding those of fish fed continuously to satiation. At the end of the experiment the percentage liver lipid content of restricted-satiation fed fish was higher than in fish fed to satiation. These differences in energy deposition patterns were, however, not sufficiently large to produce marked differences at the whole animal level, since whole body lipid energy: protein energy ratios and energy utilization efficiencies were similar for restricted-satiation and satiation fed fish.     

Cortisol effects on aerobic and anaerobic metabolism, nitrogen excretion, and whole-body composition in juvenile rainbow trout.

The influence of chronic cortisol elevation on metabolism, body composition, and fuel use patterns was examined in juvenile rainbow trout (Oncorhynchus mykiss). Measurements were performed in a control group (day 0) and in two experimental groups at days 3, 10, and 30 after treatment with a cortisol implant or a sham implant. All fish were fed 1% daily ration. Measured plasma cortisol levels were highest at day 3 and returned close to normal values by day 30 in cortisol-implanted fish. No plasma cortisol elevation was observed in the sham group. Growth was depressed in the cortisol-treated fish. Cortisol elevation resulted in increased plasma glucose concentrations during the entire experimental period, elevated CO2 production at day 3 and 30, and an elevated respiratory quotient (RQ) exceeding 1.0 on these days. Nitrogen excretion, estimated as the sum of ammonia-N plus urea-N excretion, and the nitrogen quotient exhibited small decreases at day 30. Total-N excretion, measured with a nitrogen oxidizer, was approximately twice the sum of ammonia-N plus urea-N excretion but exhibited a similar trend. Aerobic metabolism (routine O2 consumption) was higher on day 10 compared to sham-implanted fish, although not relative to day 0 control levels. Anaerobic metabolism increased substantially, as evidenced by pronounced plasma lactate elevations at days 3 and 10, a small increase in whole-body lactate on day 10, and the elevated RQ on days 3 and 30. Body composition exhibited an increase in total carbohydrate at days 3 and 10, mainly reflecting increased glycogen levels. Protein concentration was stable, indicating, in accord with the respirometry data, that protein usage did not fuel the increased metabolism or carbohydrate elevation. Redirection of nutrient uptake from food and/or mobilization of lipid stores (which decreased relative to the control group but not relative to shams) are suggested as possible energy sources for these actions of cortisol.     

Consequences of acute stress and cortisol manipulation on the physiology, behavior, and reproductive outcome of female Pacific salmon on spawning grounds

Life-history theory predicts that stress responses should be muted to maximize reproductive fitness. Yet, the relationship between stress and reproduction for semelparous salmon is unusual because successfully spawning individuals have elevated plasma cortisol levels. To tease apart the effects of high baseline cortisol levels and stress-induced elevation of cortisol titers, we determined how varying degrees of cortisol elevation (i.e., acute and chronic) affected behavior, reproductive physiology, and reproductive success of adult female pink salmon (Oncorhynchus gorbuscha) relative to different states of ovulation (i.e., ripe and unripe). Exhaustive exercise and air exposure were applied as acute stressors to manipulate plasma cortisol in salmon either confined to a behavioral arena or free-swimming in a spawning channel. Cortisol (eliciting a cortisol elevation to levels similar to those in post-spawn female salmon) and metyrapone (a corticosteroid synthesis inhibitor) implants were also used to chemically manipulate plasma cortisol. Cortisol implants elevated plasma cortisol, and impaired reproductive success; cortisol-treated fish released fewer eggs and died sooner than fish in other treatment groups. In contrast, acute stressors elevated plasma cortisol and the metyrapone implant suppressed plasma cortisol, but neither treatment significantly altered reproductive success, behavior, or physiology. Our results suggest that acute stressors do not influence behavior or reproductive outcome when experienced upon arrival at spawning grounds. Thus, certain critical aspects of salmonid reproduction can become refractory to various stressful conditions on spawning grounds. However, there is a limit to the ability of these fish to tolerate elevated cortisol levels as revealed by experimental elevation of cortisol.     

Effect of feeding frequency on food intake and growth of Arctic charr, Salvelinus alpinus L.

Social interactions and dominance hierarchy effects are important factors governing rates of growth of Arctic charr, Salvelinus alpinus L. The effects of hierarchy were increased as access to food became more restricted, i.e. feeding frequency was reduced, but these effects could not be attributed to direct competition for food since fish were fed to satiation at each feeding period. The results suggest that, whilst some fish on the restricted feeding regime were able to maintain good rates of growth, feeding by the majority of the fish was inhibited by the presence of larger individuals. Due to the importance of these hierarchy effects it was not possible to demonstrate physiological adaptations in fish allowed infrequent access to food.     

Effects of feeding regime on food consumption, growth rates and tissue nucleic acids in juvenile Arctic charr, Salvelinm alpinus, with particular respect to compensatory growth

Juvenile Arctic charr responded to a change from restricted to satiation feeding by showing a growth spurt (compensatory growth). During this period of rapid growth the fish became hyper-phagic and in the days immediately following transfer from restricted to satiation feeding showed improved food conversion efficiency compared to their counterparts raised on a liberal feeding regime. Tissue (liver and muscle) nucleic acid concentrations were influenced by feeding regime, and RNA : DNA ratios were low in both starved fish and those fed restricted rations. Following transfer from restricted to satiation feeding, tissue RNA : DNA ratios were rapidly restored to initial levels. The uses of tissue RNA: DNA ratios both in evaluating nutritional status and as growth indices are discussed.     

Laboratory studies on food intake, growth and food conversion of young herring, Clupea harengus (L.)

Food intake, growth and food conversion of young, O-group herring were studied at two temperatures and feeding regimes over a period of 19 weeks. The food intake of fish fed to satiation twice daily showed considerable variation. Food intake per fish at 14.5° C was about three times that at 6.5° C, and was generally much higher than in most other species of fish studied. The mean increase in wet weight over the 19-week period was 0.581 g/week at 14.5° C and 0.236 g/week at 6.5° C in fish fed to satiation and 0.094 g/week at 6.2° C and a ration of 1.3% of the body weight. Growth depensation was found to occur even in fish fed to satiation. The changes in specific growth rate, that is the percentage increase in weight/day, showed similar trends at different temperatures and food regimes. The mean conversion efficiency of fish on a ration of 1.3% at 6.2° C was higher than that of fish fed until satiation, at 14.5 and 6.5° C. The conversion efficiency of fish fed to satiation at 14.5° C showed a distinct decrease with increasing weight while at 6.5° C such a clear trend was not observed. In general, the conversion efficiency of young herring were found to be much lower than that of most other species studied. The weight exponent of the quantitative relationship between food intake and body weight at 14.5° C was 0.744. The total metabolic expenditure at 14.5° C, calculated using Winberg's (1956) 'utilization coefficient, gave a weight exponent of 0.773.     

Modifying the physiological responses to handling stress in beluga sturgeon Huso huso (Linnaeus, 1758); Interactive effects of feeding time and dietary fat

The present study investigated the interaction of feeding times with two dietary fat levels on physiological responses to handling stress in juvenile Beluga sturgeon. Fish were fed with two different diets (high energy; HE: 24% fat and low energy; LE: 12% fat) for 8 weeks at two feeding times; 09:00 and 16:00 (during the day) or 21:00 and 04:00 (during the night). At the end of the trial, blood samples were taken and the resting fish were held in a net out of water for 90 s as the handling stressor. Three hours after application of stress, post‐stress blood was taken. Cortisol, glucose, and lactate concentrations were considered as stress indicators. The mean values of cortisol and lactate levels did not reveal a significant difference between pre‐ and 3‐hr post‐stress samples, but the average concentration of glucose showed a significant difference. Cortisol, lactate and glucose concentrations were not influenced by the difference in the diets. Moreover, the cortisol and lactate concentrations were not affected by the different feeding times, while glucose levels were significantly affected by the feeding times with the lowest level in fish fed during the day. With respect to stress indicators, the results revealed that feeding times affected pre‐ and post‐stress secondary response to handling stress, but the rate and magnitude of metabolites (cortisol, glucose and lactate) were not affected by dietary fat levels. Therefore, it is necessary to examine the best feeding times with the interaction of feed ingredients in sturgeon farming.     

Interindividual variations in feeding and growth in rainbow trout during restricted feeding and in a subsequent period of compensatory growth

Compensatory growth responses of rainbow trout Oncorhynchus mykiss were studied by examining food intake and growth of individual fish held within groups that were switched between regimes that involved full and restricted feeding. Restricted feeding led to marked interindividual variability in food intake, probably as a result of the establishment of feeding hierarchies. This disparity in food acquisition was reflected in highly heterogeneous growth amongst the fish fed low rations. When fish were transferred from restricted to full rations, they became hyperphagic and displayed high rates of growth. Growth compensation was most marked amongst those fish which had shown the poorest growth during the period of feed restriction. These results suggest that the feeding hierarchies established under feed restriction did not persist, but were rapidly broken down when food became increasingly available, enabling the previously suppressed fish to gain access to food and to display rapid growth.     

Effect of winter feeding and starvation on the growth performance of young‐of‐year (YOY) great sturgeon,Huso huso

The objective of this study was to evaluate the feeding rate of the great sturgeon (Huso huso) young of the year (YOY) and to investigate the effects of different feeding rates in maintaining the weight of fish during short periods of winter starvation. Six feeding rates of 0.2, 0.4, 0.6, 0.8, 1.0% body weight (BW) day^?1 and feeding to satiation were considered for the first experiment. Each feeding rate was randomly assigned to three replicate tanks, with continuous feeding throughout a 5‐week winter period of water temperatures below 10°C. Fifteen fish were held in each of 18 tanks with an average initial body weight of 219.6 ± 6.9 g. After 5 weeks of feeding, the best performance was observed in fish fed 1% BW day^?1, but negative growth was observed in fish fed 0.2% BW day^?1. In the second experiment, fish were deprived of feed for 3 weeks at winter temperatures. Weights and condition factors of all fish decreased during starvation, while the differences in mean weight before and after the starvation period were not significant in fish fed a level of 0.2% BW day^?1 and those fish fed to satiation. No mortality was recorded in either experiment. Results of this study indicate that a feeding rate of 1% BW day^?1 would be sufficient for commercial fish farming of YOY of this species to maintain them over winter. Also, to maintain fish weights and prevent weight loss in overwintering ponds, a feeding rate of around 0.3% BW day^?1 seems appropriate for hatcheries.     

Energetic costs at different levels of feeding in juvenile cod, Gadus morhua L.

The energetic costs associated with feeding by juvenile cod were determined by means of an open-circuit respirometer. Fish acclimated to several temperatures (7, 10, 15 and 18°C) were kept at natural lighting levels, and fed inside their individual respirometers. They consumed a diet compounded from natural foods, at five different ration levels, their oxygen consumption being monitored continually over an 11–16 day period.
After each meal the rate of oxygen consumption increased to above the pre-feeding level, reaching a peak 8–10 h later. With each successive meal the oxygen consumption showed a cumulative increase, reaching a maximum usually after the last meal.
The elevation in metabolic rate associated with feeding was dependent upon ration size, increasing linearly as the food intake increased. The effect was also dependent upon temperature; for fish fed to satiation the total energy cost was equivalent to 11.9, 10.9, 16.4 and 17.1% of the ingested energy at 7, 10, 15 and 18°C respectively. For resting satiated fish the rate of oxygen consumption was close to the maximum rate for active fish.     

Influences of photoperiod and alternate days of feeding on plasma growth hormone and thyroid hormone levels in juvenile rainbow trout

The diurnal patterns of plasma growth hormone (GH), thyroid hormone and cortisol concentrations in rainbow trout Oncorhynchus mykiss held under three photoperiod (L : D) regimes (6 : 18, 12 : 12 and 18 : 6), and fed either daily (DF) or on alternate days (ADF) with 2·0% body mass per day of a commercial trout diet were determined. The ADF groups had reduced total mass gain and specific growth rates compared with DF fish, but photoperiod had no affect on growth for either of the feeding regime groups. In the ADF groups, the mean 24 h plasma thyroxine (T₄) and triiodothyronine (T₃) concentrations were significantly lower, both on days of feeding and days of fasting, than in DF fish held under all three photoperiod regimes, but for GH, only the 18L : 6D DF group was higher than the comparable ADF groups. There were no significant differences in mean 24 h plasma cortisol concentrations of DF and ADF groups. Diurnal patterns of plasma GH, cortisol, T₄ and T₃ were found in DF fish held under all three photoperiod regimes. Increases in plasma cortisol changes were associated with the onset of the light phase; elevations in plasma GH and T₄ concentrations were more closely associated with clock time, regardless of photoperiod; increases in plasma T₃ concentrations were strongly associated with time of feeding. In ADF groups, these diurnal changes in plasma GH, T₄ and T₃ concentrations were suppressed for both the fed and fast days, and plasma cortisol concentrations were suppressed on the fasting day. The observations are discussed in terms of the proposed anabolic, catabolic and growth regulating roles of these hormones in different growth and metabolic modifying situations in teleosts.     

Individual variations and interrelationships in feeding rate,growth rate,and spontaneous activity in hybrid tilapia (Oreochromis niloticus × O. aureus) at different feeding frequencies

A 30‐day growth trial was conducted to investigate the individual variations in feeding rate, growth rate, spontaneous activity, and their possible interrelationships in hybrid tilapia (Oreochromis niloticus × O. aureus) reared individually at feeding frequencies of one and two meals daily. Results show that the feeding rate in energy (FRe), specific growth rate in energy (SGRe), and food conversion efficiency (FCE) of fish fed twice a day were significantly higher than that of fish fed only once. However, no significant differences in distance or in time spent swimming were observed between the groups. SGRe was positively correlated with FRe in fish fed only once daily, and SGRe was positively correlated with food conversion efficiency in energy (FCEe) between the two groups. SGRe was only inversely correlated with the distance that fish swam when fed once daily. The results suggest that the individual growth differences of hybrid tilapia could be attributed mainly to variations in FRe, and that the energy costs of spontaneous activity play an important role in the differences at feeding frequency of one meal a day. However, at feeding frequencies of two meals a day, individual growth differences in fish may be caused by variations in FCEe, and energy costs of spontaneous activity have only marginal effects on the differences.     

Evidence of differences in growth and food intake regulation in different genetic strains of channel catfish

Norris and USDA-103 strains of channel catfish Ictalurus punctatus were compared for growth rate and food conversion ratio under satiation feeding and restricted feeding (1% body weight day⁻¹) regimes. At the start of the experiment Norris fish weighed 2·8 g, USDA-103 fish weighed 14·0 g. Therefore, a regression of the loge of specific growth rate against the log_e of mean body size with an empirically derived fixed slope of -0·37 was used to compare growth rates. Under both feeding regimes the USDA-103 strain had faster specific growth rates and more efficient food conversion. In subsequent studies, voluntary food intake of size matched fish (60 g average) from these two strains was compared using a radiographic method. Fish were acclimatized to tank conditions for 3 weeks prior to voluntary food intake measurement. Half of the groups were deprived of food for 2 days prior to food intake measurement, while the remaining groups were fed 1% body weight day⁻¹. The USDA-103 strain fish ate significantly more food and grew faster than the Norris strain fish. Previously fasted Norris fish subsequently ate more than their fed counterparts, whereas the fed USDA-103 fish consumed more food than the fasted USDA-103 group. When the USDA-103 strain fish were deprived of food for 4 , 2 or 0 days, all groups subsequently consumed between 4·5 and 5·0% of body weight in one meal. The USDA-103 fish, unlike the Norris fish were not stimulated to consume more after short-duration fasting. Taken together, these results suggest that there are genetic differences in growth, food conversion ratio and regulation of food intake between Norris and USDA-103 strains.     

Fasting modulates metabolic responses to cortisol, GH and IGF‐I in Arctic charr hepatocytes

Hepatocytes in primary culture from fed and 2 month fasted Arctic charr Salvelinus alpinus were exposed to physiological doses of either cortisol, salmon growth hormone (GH), salmon insulin‐like growth factor‐I (IGF‐I) or a combination of salmon GH and salmon IGF‐I. Fasting significantly lowered medium glucose levels compared to the fed fish, but had no significant effects on hepatocyte glycogen content or on the activities of enzymes involved in the intermediary metabolism. Cortisol treatment had no effect on hepatocyte glycogen content or on the enzyme activities investigated, but resulted in a significant increase in medium glucose concentration in hepatocytes isolated from fasted, but not fed fish. GH and IGF‐I treatments, both singly and in combination, significantly increased the glycogen content of hepatocytes isolated from fed fish, with less pronounced effects on hepatocytes isolated from fasted fish. The combination of GH and IGF‐I significantly increased lactate dehydrogenase activity regardless of the feeding state and significantly reduced the phosphenolpyruvate carboxykinase activity and medium glucose concentration in hepatocytes isolated from fed fish. Further, GH and IGF‐I significantly increased the activities of alanine aminotransferase and aspartate aminotransferase in hepatocytes isolated from fasted fish, but not fed fish. There were no effects of GH, IGF‐I, or their combination, on glucose 6‐phosphate dehydrogenase or 3‐hydroxyacyl‐CoA dehydrogenase activities. The results demonstrated that nutritional status of the animal modulates hepatocyte responsiveness to metabolic hormones, and suggested a role for GH and IGF‐I in hepatic glycogen conservation.     

Fasting modifies Aroclor 1254 impact on plasma cortisol,glucose and lactate responses to a handling disturbance in Arctic charr

Integrated effects of polychlorinated biphenyl (PCB) and nutritional status on responses to handling disturbance were investigated in the Arctic charr (Salvelinus alpinus). The fish were orally contaminated with Aroclor 1254 and held either with or without food for 5 months before they were subjected to a 10-min handling disturbance. Food-deprived fish were given 0, 1, 10 or 100 mg PCB kg(-1) and the fed fish 0 or 100 mg PCB kg(-1). Plasma cortisol, glucose and lactate levels were measured at 0 (pre-handling), 1, 3, 6 and 23 h after the handling disturbance. Food-deprived control fish had elevated plasma cortisol levels compared with fed fish before handling. These basal cortisol levels were suppressed by PCB in food-deprived fish, and elevated by PCB in fed fish. The immediate cortisol and glucose responses to handling disturbance were suppressed by PCB in a dose-dependent way in food-deprived fish. Although these responses were also lowered by PCB in the fed fish, the effect was much less pronounced than in food-deprived fish. There were only minor effects on plasma lactate responses. Our findings suggest that the stress responses of the Arctic charr are compromised by PCB and that the long-term fasting, typical of high-latitude fish, makes these species particularly sensitive to organochlorines such as PCB.     

An experimental investigation of the non-reproductive, post-mature state in winter flounder

Winter flounder from known locations were sorted according to their reproductive status about 4 weeks before spawning. Five groups were then fed to satiation and three groups were not fed. The experiment was terminated at the end of the summer feeding season. At termination, only one of the recently spawned fish with restricted feeding showed gonad recrudescence. Conversely, all previously reproductive fish which had been fed to satiation showed gonad recrudescence. Fish fed to satiation included some which had been initially designated non-reproductive post-mature. These experiments indicate that a non-reproductive post-mature state is inducible and may be reversible, with the switch related to nutritional status.     

Daily patterns of mRNA expression of two core circadian regulatory proteins, Clock2 and Per1, and two appetite-regulating peptides, OX and NPY, in goldfish (Carassius auratus)

The aim of this study was to examine the daily hypothalamic mRNA expression profiles for two core circadian regulatory proteins, CLOCK2 and PER1, and for two neuropeptides that regulate wakefulness and food intake, OX and NPY, in goldfish. The profiles were determined for fish at different nutritional states (i.e. fed or unfed on sampling day) and held at different photoperiods (i.e. 16L:8D photoperiod vs. constant light LL). Our results show that under a 16L:8D photoperiod, both fed and unfed goldfish exhibit clear antiphasic daily rhythms of hypothalamic Clock2 and Per1 mRNA expression levels, whereas under LL, daily Clock2 rhythms are seen in both fed and unfed fish while significant rhythms of Per1 mRNA expression only persist in unfed fish. In fish held under 16L:8D, but not under LL, there was significantly higher Per1 expression in fed fish at feeding time than in unfed fish. Daily variations in hypothalamic OX mRNA expression levels with peaks observed prior to both feeding time and the onset of darkness, were displayed under a 16L:8D photoperiod, whereas exposure to LL resulted in lower expression levels with no significant daily variations. Fish held under LL, but not under 16L:8D, showed significant daily variations in NPY mRNA expression with a peak prior to feeding time. Taken together, our results suggest that the mRNA expression of both appetite-regulating and circadian proteins display daily variations and that these patterns can be affected by external cues such as feeding and photoperiod.