Macronutrient self-selection through demand-feeders in rainbow trout

The objective of this research was to test the ability of rainbow trout to feed on, and select from, three "pure" macronutrient diets formulated to contain only one macronutrient (protein, fat, and carbohydrate). The three diets were offered to fish in three different self-feeders, the fish having to choose from them in order to compose a nutritionally balanced diet. A total of 12 trout (115 g initial body weight) were held individually in 57 liter tanks at a constant 14 degrees C. The results demonstrated that the trout were capable of composing their own complete diet, with a higher preference for protein than fat and carbohydrate. Food intake was regulated to balance energy intake and maintain steady growth. Trout showed a strict diurnal feeding rhythm, which free run under constant light with an endogenous period ranging from 18.2 to 27.7 h. These results provide the first insight into macronutrient self-selection in trout and valuable information on their feeding preferences, which may be taken into consideration when investigating fish nutrition, feeding regulation, and the design of adequate diets.     

Feeding in response to insulin but not to 2-deoxy-D-glucose in the hamster

Adult hamsters did not increase their food intake in response to subcutaneous injection of 2-deoxy-D-glucose (2DG) at doses of 50, 200, 350, 500, 750, 1,000, or 1,500 mg/kg. The failure to feed in response to 2DG was apparent regardless of time of testing and length of test. Hamsters did display marked hyperglycemia after 2DG treatment. Therefore, hamsters do detect and form neural responses to 2DG. In contrast to 2DG, injection of regular insulin, 2, 4, or 8 U/animal, caused hamsters to significantly increase their food intake over a 5-h test period. Furthermore, injection of 4 U of protamine zinc insulin twice daily also caused pronounced increases in food intake and weight gain by hamsters. Because hamsters do not seem to posses a feeding response to 2DG-induced glucoprivation, it seems unlikely that insulin elicits feeding via the glucoprivic control in this species. The results suggest that 1) hamsters lack the glucoprivic control of feeding behavior as exercised by 2DG, 2) 2DG and insulin treatments are not equivalent ingestive challenges, and 3) insulin-induced feeding may result in part from activation of a nonglucoprivic control of food intake.     

Feeding and macronutrient selection patterns in rats: adrenalectomy and chronic corticosterone replacement

To analyze further the role of corticosterone (CORT) in the control of feeding behavior, we examined the impact of adrenalectomy (ADX) and chronic CORT implants on the food intake and macronutrient self-selection patterns of adult male rats at different periods of the diurnal cycle. Consistent with a separate study of acute CORT injection in ADX rats (Kumar and Leibowitz, 1988), the present findings indicate that ADX significantly attenuated the rats' daily (24 hr) ingestion of all three macronutrients, namely, protein, carbohydrate and fat. However, food intake in the dark cycle, specifically during the first few hours after dark onset, was significantly more affected (-70%) than feeding in the later dark and light periods (-25%). Moreover, during this early dark time when circulating CORT level normally peaks, ADX appeared to have its strongest suppressive effect on carbohydrate ingestion. Chronic subcutaneous CORT implants in the ADX animals reversed these effects of surgery and generally restored the rats' eating patterns to that of the cholesterol-implanted SHAM animals. These findings suggest that CORT exerts a decisive influence on caloric intake, on the diurnal pattern of feeding, and on appetite for specific macronutrients. The impact of CORT on carbohydrate intake is apparent specifically during the active eating period, particularly at dark onset when endogenous CORT levels normally peak and carbohydrate is exhibited as the preferred macronutrient.     

Dietary self-selection and intake of protein and energy is altered by the form of the diets

The technique of dietary self-selection has been widely used to investigate proposed mechanisms of protein and carbohydrate regulation and to substantiate theories relating nutrients, brain chemistry and behaviour. These experiments have produced conflicting results which might be accounted for by differences in experimental variables or in the different techniques used to measure self-selection. The possibility that the sensory qualities of diets might alter dietary self-selection was examined in three groups of rats self-selecting protein and carbohydrate from pairs of isocaloric diets in which the physical form (finely-milled casein/corn starch powder, granular casein/corn starch powder and gelled finely-milled casein/corn starch), but not the nutrient composition of each pair was varied. In addition, since the serotonergic drug fenfluramine has been frequently used to influence dietary selection, the effects of this drug on the selection of protein and carbohydrate from the three diets were also examined. The selection of particular diets and consequently protein and carbohydrate intake was markedly altered by the form of the diet. Furthermore, the effects of fenfluramine on diet selection were also modified significantly by the form of the diet. These results suggest that intake of certain nutrients is not strictly regulated at a particular fixed value and that drug-induced changes of self-selection behaviour must be interpreted cautiously.     

Effect of melatonin on total food intake and macronutrient choice in rats

Melatonin, a hormone secreted in a rhythmic manner over 24 h mainly by the pineal gland, is used to alleviate the symptoms of jetlag and treat sleeping problems. The objective of the present study was to examine the effects of a 7-h phase-shift from the natural peak of melatonin secretion on total food intake and macronutrient selection. Forty-eight adult Wistar rats of both sexes were divided in three dietary groups, each group offered a simultaneous and different choice of a carbohydrate- and a protein-rich diet. Macronutrient intakes following intraperitoneal administration of four doses of melatonin (3000, 6000, 10 000 and 15 000 pg/ml blood) at dark onset were examined. Melatonin increased short- (4 h postinjection) and long-term (12 h postinjection) nocturnal total food intake in both male and female rats, mainly with the two highest doses. This effect of melatonin was mainly due to a short-term increase of intake across all carbohydrate-rich diet preparations (dextrin/cornstarch, cornstarch, and sucrose/cornstarch) and across genders. This consistent effect of melatonin on the intake of carbohydrate-rich diets with contrasting sensory attributes rules out the possibility that melatonin acts on sensorymotor pathways, thus suggesting that melatonin's effect on food intake is controlled by the carbohydrate content of the diet. In contrast, melatonin could be affecting some sensory or motor processes peculiar to the ingestion of protein since it increased protein-rich diet intake inconsistently across the various preparations (casein, soy isolate, and egg protein) as well as genders. This evidence supports the view that melatonin acts as a time indicator, reinforcing the animals with a “night cue”, and favors predominant carbohydrate intake normally occurring at the beginning of the activity period.     

Galanin in the PVN increases nutrient intake and changes peripheral hormone levels in the rat

In self-selection feeding paradigms, rats display differential patterns of nutrient (protein, carbohydrate or fat) intake. Factors known to influence this selection include brain peptides as well as circadian parameters. In this series of experiments we investigated the role of PVN galanin in nutrient intake during the early and late dark periods in the rat. Rats were allowed to select between three isocaloric diets enriched in protein, carbohydrate or fat. Following a 2-week adaptation period, the animals' 24-h intake was monitored for 4 weeks. Galanin was injected into the PVN and food intake was measured 1, 2 and 24 h post-injection. Galanin significantly increased the 1 h total food intake but it failed to increase the intake of any particular nutrient. Galanin had no effect 2 or 24 h post-injection. Analysis of the data grouped by preference based on the rats 24 h baseline selection patterns over the 4-week period revealed that galanin seem to increase the preferred nutrient. That is, galanin preferentially increased the intake of the carbohydrate- or fat-rich diet in animals with high (over 40% of the total food intake) 24-h baselines in this particular nutrient. Finally, analysis of the plasma hormone levels after paraventricular galanin administration revealed a significant increase in noradrenaline levels, a small reduction in plasma insulin with no effects on adrenaline, glucose or corticosterone. The data revealed that galanin in the PVN influences both food intake and metabolic functioning. PVN galanin significantly increases sympathetic outflow and seems to stimulate the intake of the individual rat's preferred macronutrient.     

Deficits in the control of food intake after hypothalamic paraventricular nucleus lesions

Noradrenergic mechanisms of the hypothalamic paraventricular nucleus (PVN) have been shown to play an important role in the stimulation of feeding To determine the influence of this nucleus in monitoring and controlling responses to physiological and pharmacological challenges, PVN electrolytic lesion rats were tested for their behavioral responsiveness to agents known to affect the alpha-2 noradrenergic system as well as release of corticosterone, and to short- and long-term periods of food deprivation. Discrete lesions of the PVN produced enhanced feeding, particularly of carbohydrate, in freely-feeding rats maintained on a macronutrient self-selection paradigm. Lesion rats demonstrated a behavioral deficit in food intake regulation (a decrease in carbohydrate ingestion) in response to 5-hr and 24-hr fasts, showed a disturbance in circadian feeding, and exhibited a dramatic decrease in circulating corticosterone. However, feeding in response to 2-deoxy-D-glucose and insulin remained intact, suggesting that noradrenergic receptors within the PVN are not involved in the mediation of glucoprivic-induced feeding.     

The role of a noxious taste in determining food intake in the rat

Adult female rats were given access to cornstarch, fat, and 2 cups of casein ad lib. Sucrose octaacetate was added to one or both casein cups in concentrations of 0, 0.1, or 2.5%. The lower concentrations of SOA had no effect on total caloric intake or food choices, although a taste aversion experiment indicated that the rats could detect SOA at this level. The 2.5% concentration had no effect on total caloric intake. If only one casein cup was treated with SOA, the animals consumed 70% of their protein from the unadulterated casein supply, but total protein intake was unchanged. If both cups were treated with 2.5% SOA, protein intake decreased, but carbohydrate consumption showed a compensatory increase. Fat intake was unchanged.     

Satiation due to CCK-8 derivative infusion into VMH is related to a specific macronutrient selection

Diet self-selection from carbohydrate, protein and lipid offered simultaneously was studied in adult male rats infused for 8 days with pyroglutamyl-CCK-8 (pGlu-CCK-8) (0.8 pmol/hr), a derivative of the COOH-terminal octapeptide of cholecystokinin (CCK), or vehicle bilaterally into the ventromedial hypothalamus (VMH). Infusion with pGlu-CCK-8 markedly decreased total daily energy intake mainly due to a suppressed carbohydrate intake. Both pGlu-CCK-8 and vehicle infusions caused a nonspecific decrease in protein intake, but the vehicle infusion caused a compensatory increase in lipid intake as well as carbohydrate intake, so their total energy intake was well maintained. These results suggest that the process of infusion into the VMH elicited itself a suppressive effect on protein intake, but that pGlu-CCK-8 infusion elicited a more specific suppressive effect on carbohydrate intake and, to a lesser extent, on lipid intake. These results also support the prediction that centrally administered CCK may suppress food intake and show in addition that such a suppression occurs selectively. The similarities and the differences in the patterns of macronutrient selection produced by vehicle of pGlu-CCK-8 infusions into the VMH provide further evidence of the unique functions of VMH and CCK in the feeding process.     

Macronutrient selection through postingestive signals in sea bass fed on gelatine capsules

The role of the food orosensory properties on protein (P), fat (F) and carbohydrate (CH) self-selection was investigated in fish fed gelatine capsules containing pure macronutrients. A total of 40 sea bass (39.6+/-6.2 g initial body weight) distributed in eight 75-l tanks were used. In a sequence of experimental phases, sea bass were fed a pelleted complete diet, an encapsulated complete diet or a combination of separately encapsulated pure macronutrients. In order to induce associative learning, capsules containing a given macronutrient were paired with a particular colour. Our results demonstrate that fish are able to regulate food intake, so as to balance their energy intake, when they are fed a complete encapsulated diet and, therefore, without using the orosensory properties of the diet. Moreover, sea bass learn to discriminate and select among colour-coded, pure macronutrient capsules to compose a complete and balanced diet using colour as the only external cue. The composition of selected diet was 55% P, 23% CH and 22% F in terms of macronutrient percentage. The diet orosensory properties do not seem to be necessary to regulate macronutrient intake either, suggesting that an associative learning between capsule colour and content can be established through monitoring of macronutrient intake by postingestive and/or postabsorptive mechanisms. These results provide the first insight into energy and macronutrient self-selection by fish fed on gelatine capsules containing separate macronutrients.     

The satiety effects of intragastric macronutrient infusions in fatty and lean Zucker rats

To evaluate satiety in the hyperphagic, genetically obese Zucker “fatty” (fafa) rat, food-deprived fatty and lean (FaFa) control rats were given equicaloric intragastric infusions consisting largely of fat, carbohydrate, or protein. Relative to distilled water infusion, these infusions resulted in immediate reductions of food intake in both fatty and lean rats allowed to feed 20 min post-infusion. Cumulative food intakes remained reduced throughout the 2 hr period of observation. Thus, despite its hyperphagia, the fatty rat is responsive to the satiating effect of infused nutrients. However, the relative satiating effectiveness of the macronutrient infusions differed for the two genotypes. In lean rats, the different macronutrient infusions resulted in equivalent reductions of feeding. In contrast, in fatty rats, fat was the least satiating and protein was the most satiating macronutrient. Moreover, compared to lean rats, fatty rats displayed less initial suppression of feeding after fat infusion and greater overall suppression after protein infusion. These effects are consistent with the long-term feeding behavior of the fatty rat for the different macronutrients and may be related to pre- and postabsorptive metabolic alterations that have been documented in this animal.     

Macronutrient selection through post-ingestive signals in sharpsnout seabream fed gelatine capsules and challenged with protein dilution

Sharpsnout seabream ability for macronutrient self-selection was studied using gelatine capsules containing pure macronutrients. In particular, the existence of non-oropharyngeal chemosensory pathways involved on protein (P), fat (F) and carbohydrate (CH) selection, as well as sharpsnout seabream response to dietary protein dilution were investigated. In a sequence of experimental phases, sharpsnout seabream were fed a pelleted complete diet, an encapsulated complete diet or a combination of separately encapsulated pure macronutrients. In order to induce associative learning, capsules containing a given macronutrient were paired with a particular colour. The animals composed a diet containing 62.7% P, 21.3% CH and 16.0% F, in terms of macronutrient percentage intake, and this selection pattern was maintained throughout all experimental phases. In a second experiment, individually kept sharpsnout seabream were challenged with protein dilution. After protein capsules were diluted (from 91.9% P to 56.3% P) with cellulose, the animals increased their protein intake to compensate for dilution in such a way that their energy intake was not significantly modified (17.4 kJ/100 g BW vs. 17.6 kJ/100 g BW after dilution). These results show that sharpsnout seabream feeding on encapsulated diets are able to select and maintain a particular diet composition, as well as sustain their energy intake, without using the diet's oropharyngeal chemosensory properties. Moreover, they were also able to maintain their protein intake after dilution, which highlights the importance of this macronutrient in this omnivorous species.     

Central insulin potentiates eating elicited by 2-deoxy-D-glucose

2-Deoxy-D-glucose (2DG) elicits glucoprivic food intake whether administered centrally or systemically. Insulin, on the other hand, elicits glucoprivic food intake when administered systemically but reduces food intake when administered centrally. The purpose of these experiments was to determine the interaction of centrally administered insulin with systemically administered 2DG on feeding. In the experimental condition, male Sprague-Dawley rats were administered 5 mU insulin into the third cerebral ventricle (i3vt) followed 2 h later by a subcutaneous injection of 250 mg/kg of 2DG. Contrary to expectations, third ventricular insulin significantly increased 2DG-induced hyperphagia. A replication using doses of insulin ranging from 1 to 10 mU revealed a dose-dependent response. Whereas the lowest dose of insulin (1 mU) did not reliably change food intake, doses of 2.5, 5, and 10 mU significantly enhanced 2DG-induced feeding. Consistent with previous reports, centrally administered insulin, when given alone, caused a significant reduction of 24-h body weight and chow intake. To assess if the insulin-induced hyperphagia was a result of leakage from the ventricles, we peripherally administered 5 mU of insulin and observed, if anything, a slight decrease of food intake. These studies suggest that in the presence of central glucoprivation, a distinct anabolic action of centrally administered insulin overrides the normally observed catabolic response and increases the hyperphagic feeding response induced by 2DG.     

Effect of starvation or restriction on self-selection of macronutrients in rats.

Two models of food deprivation were used to study feeding behavior: starvation and dietary restriction. Rats starved for 3 days had decreased protein intake during the first 2 days of refeeding followed by increased carbohydrate consumption compared to controls. During refeeding, total intake was initially low compared to controls. In a second starvation study of similar design, brain tissues [raphe, ventromedial hypothalamus (VMH)] and sera were collected for analysis before refeeding and on day 2 of refeeding. Starved and starved-refed rats had increased serum beta-hydroxybutyrate versus controls. In rats restricted for 5 days (5 g/day), total food intake was increased immediately and was characterized primarily by carbohydrate intake. Serotonin levels in the raphe were decreased in restricted rats and 5-hydroxyindole acetic acid (5-HIAA) increased in restricted-refed rats. Restriction caused an increase in blood levels of beta-hydroxybutyrate and a decrease in insulin and glucose compared to controls. Fat selection remained low throughout all studies. The data suggest that starvation and food restriction elicit different patterns of macronutrient selection upon refeeding.     

Feeding induced by GABA(A) receptor stimulation within the nucleus accumbens shell: regional mapping and characterization of macronutrient and taste preference.

This study investigated the areas of the nucleus accumbens shell involved in the modulation of feeding behavior by GABAergic stimulation and characterized this response using macronutrient diets as well as saline, sucrose, and saccharin solutions. The GABA agonist muscimol induced a pronounced feeding response when infused in the medial nucleus accumbens shell but not in the ventral or lateral accumbens shell. In the macronutrient preference study, muscimol increased the intake of both high fat and high carbohydrate diets when presented separately. When both diets were available simultaneously, muscimol stimulated feeding of both diets to the same degree. Muscimol elicited a robust increase in the consumption of sucrose solution. However, no effect of muscimol was demonstrated for water, saline, or saccharin intake. These findings provide evidence for a selective role for GABA-sensitive neurons in the medial accumbens shell in the regulation of ingestive behavior and further suggest that GABA(A) receptors in this region do not modulate palatability, macronutrient selection, or rewarding properties of food.     

Pancreatic glucagon's effects on satiety and hepatic glucose production are independently affected by diet composition

We tested whether pancreatic glucagon's effects on satiety and hepatic glycogenolysis depend on variations in the macronutrient composition of the diet. Rats were kept on high carbohydrate, high fat, or high protein diets (HC-, HF- or HP-rats, respectively) and adapted to a 5 hour feeding (11:00-16:00)-19 hour deprivation (16:00-11:00) schedule. Glucagon (540 mcg/kg body weight) was intraperitoneally injected at 14:00. Glucagon stimulated similar amounts of hepatic glycogenolysis in HC- and in HF-rats and less glycogenolysis in HP-rats, but glucagon decreased food intake only in HC-rats. When HF- or HP-rats were prefed the HC-diet once for 1 hour (11:00-12:00) prior to injection, glucagon stimulated hepatic glycogenolysis similarly in both groups but decreased food intake only in HF-rats. Therefore, stimulation of hepatic glycogenolysis by glucagon is not sufficient to elicit satiety under all feeding conditions. The results also suggest that glucagon-induced satiety is not limited to carbohydrate intake.     

Photoperiod modulates torpor and food intake in Siberian hamsters challenged with metabolic inhibitors

Siberian hamsters, Phodopus sungorus, exposed to a short photoperiod (SP) were challenged with 2-deoxy-D-glucose (2DG), which disrupts glycolysis and induces torpor in animals maintained in long photoperiods (LP), or mercaptoacetate (MA), which disrupts fatty acid oxidation. SP decreased rather than facilitated the induction of torpor by 2DG; decreased torpor in response to 2DG coincided with onset of spontaneous torpor in SP hamsters. In contrast, MA induced hypothermia in hamsters kept in SP but not LP, but did not induce full torpor. We conclude that short day lengths do not induce spontaneous torpor by increasing responsiveness to glucose availability; instead, responsiveness to glucoprivation is, if anything, blunted in SP. The occurrence of spontaneous torpor may be unrelated to concurrent metabolic fuel availability. Although 2DG and MA had different effects on thermoregulation in short vs. long day lengths, each treatment decreased 24-h food intake in hamsters in both photoperiods.     

The NMDA receptor antagonist MK-801 alters lipoprivic eating elicited by 2-mercaptoacetate

Eating behavior is controlled, at least in part, by levels of circulating metabolic fuels such as glucose and free fatty acids, and drugs that interfere with the availability of these fuels can elicit eating. One such drug is 2-mercaptoacetate (2MA), an inhibitor of fatty acid oxidation. Evidence also suggests that NMDA receptors may mediate some aspects of normal eating and satiety. The present study was conducted in order to determine whether NMDA receptors may play a role in feeding elicited by 2MA. Rats received intraperitoneal injections of either saline, 2MA, the non-competitive NMDA receptor antagonist MK-801 or a combined injection of 2MA and MK-801, and subsequent intake of a fat-enriched, mash diet was measured at 1, 2, 3 and 4 h post-injection. Results showed that cumulative food intake was significantly increased by 2MA alone, as compared to saline controls, with most of the 2MA-elicited eating occurring during the first hour post-injection. While MK-801 alone did not alter food intake, it did have a biphasic effect on feeding elicited by 2MA. MK-801 initially suppressed and later enhanced eating elicited by 2MA. Although it is unclear whether MK-801 is acting centrally, peripherally or both to alter 2MA-induced eating, these results implicate NMDA receptors and the neurotransmitter glutamate in the regulation of lipid-associated eating and satiety.     

Test-meal palatability alters the effects of intragastric fat but not carbohydrate preloads on intake and rated appetite in healthy volunteers

Manipulations of test meal palatability and nutritional need-state to examine feeding behaviour have, to date, been studied in isolation. Recent investigations have attempted to examine these influences in combination. In the present study, healthy young males received intragastric infusions of soup (265 or 1514 kJ) on four different occasions. The infusion was shortly followed by a meal varying in its palatability (PALATABLE or BLAND). The effect of macronutrient type (CHO or Fat) in the high-energy preloads was also examined in a between-subject manner. High CHO preloads significantly decreased test meal intake and this decrease was unaffected by meal palatability. High fat preloads did not significantly reduce test meal intake. Additionally, more food was consumed following high fat preloads when the test meal was PALATABLE. Within-meal ratings of appetite revealed that hunger was diminished to a greater extent by CHO than by Fat preloads. Appetite was stimulated by the PALATABLE meal to a greater extent in the group receiving Fat than in those receiving the CHO preload. Comparison with a similar oral preloading study revealed differences that suggest possible interactions between cognitive, oro-sensory and gastric controls of feeding when palatable foods are consumed.     

Acquisition of dietary self-selection in rats with normal and impaired oral sensation

To study the mechanisms of dietary self-selection, the dietary choice behavior of rats was observed for up to 5 weeks, beginning on the first day of exposure to two nutritionally different diets, a high-protein diet (44% protein) and a protein-free carbohydrate diet. In Experiment 1 normal rats selected equal amounts of the two selection diets at first and over a 7-day period gradually modified the choice ratio until a stable protein intake of 14.6% (SE,0.4) of total intake was reached. In Experiment 2 rats were subjected to partial trigeminal deafferentation, which impairs oral somatosensory input (touch, temperature, pain), before the two selection diets were introduced. The deafferented animals did not develop a stable selection pattern; their protein ratio varied over the entire possible range (0-44%) throughout the experiment. It is hypothesized that quantitative protein/carbohydrate selection involves an associative learning process in which somatosensory inputs from the feeding activity and/or from the properties of the food link dietary choice behavior to later metabolic consequences.