Hepatic vagotomy disrupts somatotropin-induced protein selection

Rats treated with somatotropin (STH) and allowed to self-select between diets varying in protein content will consume more of the high-protein diet. The objective of this study was to determine the role of the hepatic vagus nerve in this ability to select protein. Female Sprague-Dawley rats (n=40) received a hepatic vagotomy (HVAGX) or a sham surgery. Postsurgery, the rats were maintained on pelleted diets for 2 weeks, after which the rats were adapted to selecting between powdered diets with 5% casein and 30% casein. After a 7-day adaptation to diet selection, rats in each surgical treatment group were treated with STH (4 mg/day) or physiological saline for 14 days. Body weight and intake were recorded daily. STH treatment increased growth rate to a similar degree in both sham and HVAGX groups. Despite causing an increase in total food intake, there was no effect of HVAGX alone on body weight. Relative to the sham-saline group, sham-STH in treated rats had greater total food intake that was accounted for entirely by increased consumption of the 30% protein diet and no change in intake of the 5% diet. In contrast, HVAGX+STH rats exhibited 20-30% increases in consumption of both the 5% and 30% protein diets. Thus, the HVAGX+STH rats recognized an increased need for protein, but were unable to distinguish between the high- and low-protein diets and selected more of both. The data suggest that the ability to alter diet selection in response to a stimulation of protein accretion is at least partially mediated through the liver and hepatic branch of the vagus nerve.     

The effects of liver denervation on food and water intake in the rat

In Experiment 1, liver denervations or sham operations were performed on rats in two separate trials. Food and water intakes and body weights of denervated rats did not deviate significantly from their sham operated controls. Male and female rats responded similarly. In Experiment 2, in addition to daily food and water intakes, initial daily meal size was investigated in two groups of liver denervated and sham operated rats. Initial daily meal size was determined during a one hour test at the start of the dark period of the light:dark cycle. In one group of rats a chow diet was used for testing, while a liquid diet was utilized in the second group. According to the “liver glucoreceptor preabsorptive satiety and general food intake control hypothesis” the denervated rats should have experienced a depression of daily food intake and preabsorptive satiety. No differences were found in either preabsorptive satiety or daily food and water intakes and body weights when denervated rats were compared to sham operated control animals. The results question whether liver glucoreceptors have any detectable influence on the control of feeding behavior. Certainly the data does not support the role of liver glucoreceptors as a major controller of feeding behavior in the rat.     

Effects of hippocampal lesions on adaptive intake of diets with disproportionate amounts of amino acids

Bilateral double electrolytic overlapping lesions were placed in dorsal-lateral hippocampus of male 230 g rats, and their food intake responses to the ingestion of diets containing disproportionate amounts of amino acids were examined. Rats with such lesions and intact control rats maintained their normal intakes of the 6% casein basal diet or a threonine basal amino acid diet postoperatively. However, they exhibited marked initial food intake depression, similar to that of intact rats, when fed the threonine imbalanced amino acid diet. Also, animals with lesions in certain areas of the dorsal-lateral hippocampus showed facilitated adaptation to the amino acid imbalanced diet. Similar severe reduction in food intake with relative lack of adaptation were observed in both the intact controls and rats with hippocampal lesions when fed amino acid diets completely devoid of threonine. Initial food intake of rats with hippocampal lesions was inhibited drastically as was the case with the intact controls when fed a 75% casein high protein diet. All rats, either intact or lesioned, showed similar slow adaptation patterns with the prolonged ingestion of the high protein diet. The initial food intake responses and facilitated adaptation of the animals bearing lesions in certain areas of the hippocampus suggest that such areas are not crucially involved in the inhibition of food intake of rats fed disproportionate amounts of dietary amino acids. Rather, such areas of lesions in the hippocampus may play a role in a system governing the behavioral adaptation of the intake of amino acid imbalanced diets but not of diets containing amino acids in general excess. This would also indicate that different mechanisms control the intake of amino acid imbalanced diets and diets containing amino acids in excess.     

Effect of amygdaloid lesions on dietary intake of disproportionate amounts of amino acids

Male rats with bilateral electrolytic lesions in the anterior, medial or posterior aspects of the ventral amygdala and groups of intact rats were fed, in turn, basal, imbalanced or deficient amino acid diets involving threonine or isoleucine as the limiting amino acid, and then a low protein (6% casein) followed by a high protein (75% casein) diet. No change in food intake was observed in animals fed the threonine basal diet postoperatively. When the threonine or isoleucine imbalanced diet was substituted for the respective basal diet, animals with lesions in certain areas of the medial amygdala showed little or no depression in food intake of the imbalanced diets, while all other rats with amygdala lesions reduced their food intake markedly, as did intact controls, when fed such diets. All animals, however, curtailed their food intake of the deficient or high protein diets. The lack of responsiveness of the animals with medial amygdaloid lesions to the imbalanced diets suggests that these areas may be involved in a system regulating food intake of animals fed diets containing imbalanced amino acid mixtures.     

Cingulate lesions and behavioral adaptation to amino acid imbalanced diets

The effect of anterior cingulate cortex lesions on dietary intake and adaptation of disproportionate amounts of amino acids was examined. Rats with bilateral electrolytic lesions in the anterior cingulate cortex and sham-operated rats were fed, in turn, amino acid basal, imbalanced or devoid diets involving threonine and isoleucine as the growth limiting amino acids, and then a low protein (6% casein) followed by a high protein (75% casein) diet. Lesions of the anterior cingulate cortex did not prevent the initial depression in food intake of the amino acid imbalanced diets, but shortened the duration of anorexia associated with dietary amino acid imbalances. Cingulate lesions did not influence the food intake of rats fed amino acid devoid diets. When switched from a low protein to a high protein diet, animals bearing lesions and sham-operated controls reduced markedly their initial food intake and adapted to the high protein diet in similar manner. It was concluded that the initial food intake depression associated with a dietary amino acid imbalance is a direct response to postingestive cues which influence food intake. Moreover, that the difference in adaptive intakes of the cingulate cortex lesioned animals who ingested a diet of imbalanced amino acids or of high protein, indicates that separate mechanisms act to control food intake of animals fed diets containing imbalanced amino acid mixtures or diets with excessive amounts of protein.     

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.     

Dietary hyperphagia in rats: role of fat, carbohydrate, and energy content

Dietary energy, fat and carbohydrate content were varied to determine the nutritional factors responsible for hyperphagia induced by feeding rats high-fat diets. In the first experiment, rats were fed isoenergetic high-fat or high-carbohydrate diets for 2 weeks. Weight gain and energy intake were lower in rats given the high-fat diet. When some of the rats were switched to a diet that was high in fat, carbohydrate and energy, gram food intake was initially unchanged, resulting in a substantial increase in energy intake and weight gain. Energy intake gradually declined over the 4 weeks following the switch to the high-energy diet. In the second experiment, rats were fed high-fat diets that were either high or low in carbohydrate content and either high or low in energy content (kcal/g). Rats fed a high-fat diet that was high in energy and carbohydrate ate the most energy and gained the most body weight and carcass fat. In the third experiment, rats were fed high-carbohydrate diets varying in fat and cellulose content. Energy intake and body weight gain varied directly as a function of caloric density regardless of the fat or cellulose content of the diets. It is concluded that hyperphagia induced by feeding high-fat diets is not due to the high dietary fat content alone. Rather, high levels of fat, carbohydrate, and energy interact to produce overeating and obesity in rats fed high-fat diets.     

Effect of diet textural characteristics on the temporal rhythms of feeding in rats

To examine whether the diurnal rhythms of protein-rich and carbohydrate-rich diet ingestion can be altered by presenting the diets in different textural forms, adult male Wistar rats were assigned to two dietary groups. One group received a two-way choice between high-protein powder and high-carbohydrate granular (HPP-HCG) diets. In the other group the textures were reversed [high-protein granular and high-carbohydrate powder (HPG-HCP) diets]. Rats fed HPP-HCG diets selected significantly less protein (kcal) vs. rats fed HPG-HCP diets, during the 24-h and 12-h dark phase and during the 4-h early and late dark phases. Carbohydrate intakes of the two dietary groups were not significantly different. Total caloric intake for the HPG-HCP dietary group was significantly higher than that of the HPP-HCG dietary group during the 24-h and 12-h dark phase. Body weight was significantly lower in rats fed HPP-HCG diets. In conclusion, macronutrient-rich diets presented in different textural forms alter protein-rich diet ingestion and total energy intake.     

Combined pregastric and gastric stimulation by food is sufficient for normal meal size

Rats equipped with a pyloric noose ate liquid food after 3-hr food deprivation. When the noose was open, ingested food accumulated in the stomach and entered the intestine in the normal fashion. When the noose was open, rats ate a mean meal size (MS) of 8.1 ml, displayed a postprandial satiety sequence with a mean latency to rest (LR) of 16.5 min, and had a gastric fluid volume at the end of a meal that was 111.7% of the MS volume. When the noose was closed, ingested food was trapped in the stomach and did not enter the intestine. When the noose was closed, rats ate a mean MS of 8.4 ml, displayed a postprandial satiety sequence with a mean LR of 15.4 min, and had a gastric fluid volume at the end of a meal that was 148.5% of the MS volume. These results show that (1) preabsorptive stimuli from pregastric and gastric sites are sufficient for normal meal size and postprandial satiety behavior, and (2) that satiety signals produced by preabsorptive contact of food with the intestine and by postabsorptive nutrient stimuli are not necessary for normal meal size and postprandial satiety behavior under experimental conditions that resemble spontaneous feeding in the light phase of the diurnal cycle.     

Protein selection, food intake, and body composition in response to the amount of dietary protein

Though not universally observed, moderately low-protein diets have been found to increase caloric intake and body fat. It appears that animals overeat in calories in order to obtain more dietary protein. For animals to control protein intake, they must be able to distinguish between two isocaloric diets containing different percentages of protein and make the appropriate dietary selection on the basis of their previous history of protein intake. Experiment 1 examined the 24-h diet selection (5 vs. 35% casein) of Sprague-Dawley rats that had been previously fed diets containing various percentages of dietary protein (5, 10, 20, 35, or 60% casein). Animals fed 5, 10, or 20% dietary protein showed a preference for the higher protein selection diet. In contrast, no significant diet preference was found in animals pre-fed the two higher levels of dietary protein (35 or 60% casein). In this study, daily food intake and body fat of rats fed the low-protein diets (5 and 10% casein) were similar to rats fed the 20% casein diet. Experiment 2 examined the effects of the level of methionine supplementation on rats fed 10% casein. In this study, food intake and body fat were increased by approximately 20% in rats fed 10% casein diets, regardless of the level of methionine supplementation (0.3 vs. 0.15%). Together, the results suggest that the presence of low-protein-induced hyperphagia helps maintain body protein levels in the face of moderately low dietary protein and promotes an increase in the amount of body fat and energy.     

Meal composition influences subsequent food selection in the young rat

Food selection one-half hour after the ingestion (2 g) of either a protein containing or a protein-free (carbohydrate) diet was studied in young rats. Following a 12 hr fast the rats were allowed 15 minutes to consume the meal (premeal). Thirty minutes later, they had access to two isocaloric diets that differed only in protein and carbohydrate content. During the first hour of ad lib feeding, protein intake and protein concentration selected were lower in the 45% casein prefed group compared to the carbohydrate group. A further reduction in protein intake and protein concentration occurred when the protein content of the premeal was increased to 70% from 45% casein. Rats prefed with the 70% casein diet significantly reduced their daily total food intake (12 hr) compared to either the 45% casein or carbohydrate prefed group. It is concluded that both quantity and composition of food selected is influenced by the composition of the preceding meal.     

Anorexia in rats caused by a valine-deficient diet is not ameliorated by systemic ghrelin treatment

Rodents exhibit aversive behavior toward a diet that lacks at least one of the essential amino acids. We sought to determine whether the particular form of anorexia caused by such diets could be ameliorated by the administration of orexigenic peptides while simultaneously analyzing the neural mechanisms underlying anorexia. Rats were fed a valine-deficient diet, which induced severe anorexia (reducing food consumption by 80%). The severe anorexia was associated with a significant decrease in the cerebrospinal fluid valine concentration and hyper-ghrelinemia. Between 6 and 12 days after initiation of the valine-deficient diet, we injected rats twice daily with valine and/or an orexigenic peptide (ghrelin, neuropeptide Y, or agouti-related protein) either i.p. or i.c.v.. We then measured dietary intake. An i.c.v. valine injection allowed earlier food intake compared with an i.p valine injection and increased the density of c-Fos-positive ependymal cells lining the third ventricle. Whereas an i.c.v. injection of ghrelin or neuropeptide Y increased consumption of the valine-deficient diet, i.p injection of ghrelin or i.c.v. injection of agouti-related protein did not. Following i.c.v. administration of either valine or ghrelin, we did not observe complete recovery of consumption of the valine-deficient diet. This may be due to the ineffectiveness of peripheral ghrelin and central agouti-related protein and/or to conditioned aversion to the valine-deficient diet. Since ghrelin is known to be involved in food anticipatory activities, whether the hyper-ghrelinemia observed in valine-deficient rats play role in foraging behavior other than food intake is the future study to be investigated.     

Effects of dietary protein on food and water intake in spontaneously hypertensive rats

We have been studying the development of hypertension in spontaneously hypertensive rats (SHR) fed a low protein diet. The effects of a low protein diet upon food and water intake were examined. Body weight gain, food and water intake were measured in three to twenty-three week-old SHR and Wistar Kyoto rats (WKY) fed diets containing 8%, 15% or 25% casein. Body weights of SHR and WKY fed an 8% casein diet were significantly lower at 23 weeks than rats on the higher protein diets, although both groups on the 8% diet consumed more food and water per g of body weight. In addition, SHR fed an 8% casein diet drank less water per gram of food than WKY or SHR fed 15% and 25% casein diets. These results indicate that changes in food and water intake, as a consequence of low protein diets, should be an additional consideration when examining the effects of dietary protein on the development of hypertension.     

Diet selection and metabolic fuels in three models of diabetes mellitus

Dietary self-selection and circulating metabolic fuels (glucose, free fatty acids, ketones) were examined in three forms of experimental diabetes mellitus in rats: pancreatectomy and streptozotocin treatment in adult and neonatal rats. Changes in diet selection resulting from insulin replacement also were examined. Differences were found in diet selection and circulating metabolic fuels between these types of diabetes. Mildly diabetic rats selected large amounts of fat while more severely diabetic rats primarily selected protein. Insulin treatment enhanced carbohydrate intake of diabetic rats and nearly normalized diet selection and circulating metabolic fuels. All diabetic groups exhibited severe glucose intolerance. These results support the observations of the beneficial effects of low-carbohydrate diets, question the generality of the use of high-fat diets, and suggest a more important role for high-protein diets in energy regulation in severely diabetic rats.     

Adaptation to water restriction through dietary selection in weanling rats

For each of five pair of diets, 20 weanling rats were equally divided into ad lib and restricted water groups. Caloric density, palatability, as well as nutrient composition of the diet pairs were manipulated. Though lower in absolute food intake than controls, water-restricted rats took a larger proportion of their food from diets lower in protein content and higher in fat content, regardless of caloric content or palatability. The results demonstrate both a quantitative and qualitative dietary adaptation to water restriction.     

Caloric compensation following insulin administration in rats

Caloric intakes following injections of insulin were examined in male rats maintained on diets of different caloric densities. Following saline or insulin injections, food intake was measured every two hours for six hours or food was withheld for six hours until recovery of normo-glycemia and total intake was measured at seven and eight hours post-injection. Additionally, levels of plasma metabolic fuels, liver glycogen and stomach contents were measured following saline and insulin injections. Animals in both the continuous and delayed access conditions consumed the same number of calories following injections of insulin. Similarly, the increases in intake relative to saline baseline conditions were comparable across diet groups in both conditions although animals tested in the delayed access paradigm were less variable in their response. Changes in levels of metabolic fuels, stomach contents and liver glycogen were found to be consistent with the known effects of insulin and counter-regulatory responses to the induced hypoglycemia.     

Maternal malnutrition in the rat: Effects on food intake and body weight

The effects of dietary protein level on food intake and body weight were examined in adult female rats during a 35-day pre-mating period and during gestation and lactation. During the pre-mating period, no differences in daily food intake were observed among rats fed a 6% casein, 8% casein or 25% casein diet. However, during this period, rats fed the 6% casein diet gained significantly less weight than those with ad lib access to the 8% or 25% casein diets or than rats pair-fed the 25% casein diet in amounts equivalent to that consumed by rats in the 6% or 8% casein groups. Additionally, rats fed the 6% casein diet displayed decreased efficiency of energy utilization, calculated as weight gain per 100 kilocalories consumed, relative to rats fed the 8% or 25% casein diets. No differences in food intake were observed among the groups during gestation. However, rats fed the 6% casein diet gained less weight than rats fed the 8% or 25% casein diets. During lactation rats fed either the 6% or 8% casein diet consumed significantly less food than animals given the 25% casein diet ad lib. During the second week of lactation, rats receiving ad lib access to the 25% casein diet gained weight while those receiving the 6% or 8% casein diets continued to lose weight. At parturition, body weights of pups did not differ as a function of dietary condition. However, by day 12 of life, pups whose dams had ad lib access to the 25% casein diet weighed significantly more than pups whose dams consumed the 6% or 8% casein diet or whose dams were pair-fed the 25% casein diet in amounts equivalent to those consumed by rats fed the 6% or 8% casein diet.     

Sensory versus dietary factors in cafeteria-induced overweight

Normal Wistar rats became hyperphagic and obese when presented with high-fat, high-sugar or “cafeteria” diets. This phenomenon could be due to the very palatability of the offered foods but a metabolic factor could be suspected. In an attempt to distinguish the effects of the nutritional characteristics from those of the sensory properties of food, a new cafeteria diet, the “isocafeteria” diet, was developed. It consists of a choice of foods of similar, well-balanced composition but varying in taste, smell and texture. It was confirmed that in normal Wistar rats, the high-fat and the traditional “cafeteria” diet led to extra-weight gain. It was demonstrated that the daily presentation of a new choice of the palatable foods which composed the “isocafeteria diet” led also to a sustained increase in food intake and to overweight. Variety and high palatability are per se sufficient factors to overcome regulatory mechanisms. It was shown that the metabolic efficiency of such a regimen was strikingly higher than those of high-fat and traditional cafeteria diets.     

Nutritional studies of the roles of dietary protein levels and umami in the preference response to sodium chloride for experimental animals

Previously we found that appetite or preference for sodium chloride depends not only on a genetic factor, but also on the nutritional status, i.e., dietary protein levels. To determine whether all kinds of protein had reducing effects on sodium chloride intake, the effect of dietary protein source on appetite or preference for sodium chloride was studied. It was found that the effect depended not only on protein levels, but also on the protein sources, i.e., amino acid patterns of diets. On the other hand, the turnover time of gustatory taste bud cells was measured by the pulse-labeled method using 3H-thymidine. It was found that the turnover time of gustatory cells in the taste buds of rats fed a low-protein diet was longer than in rats fed a high-protein diet. This suggested that some reduction of receptor functions may occur with low-protein diet. It was found that the dietary protein levels and umami also modified appetite or preference for alcohol solutions.     

Effect of food deprivation and maintenance diet composition on fat preference and acceptance in rats

High-fat diets typically elicit greater kcal intake and/or weight gain than low-fat diets. Palatability, caloric density, and the unique postingestive effects of fat have each been shown to contribute to high-fat diet hyperphagia. Because long-term intake reflects the sum of many individual eating episodes (meals), it is important to investigate factors that may modulate fat intake at a meal. The present studies used high-fat (hi-fat) and high-carbohydrate (hi-carb) liquid diets (both 2.3 kcal/mL) to assess the effect of hunger level (0 versus 24-h food deprivation) and fat content of the maintenance diet (12 versus 48%) on fat preference (when a choice among foods is offered in a two-bottle test), and acceptance (only one food offered) in male rats. Preference for hi-fat relative to hi-carb (two-bottle test) was enhanced by 24-h food deprivation, and by a high-fat maintenance diet. In contrast, neither deprivation nor maintenance diet composition influenced relative meal size (one-bottle test) of hi-fat and hi-carb: irrespective of test conditions, meal size of hi-fat was bigger than meal size of hi-carb.