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.     

Norepinephrine and amino acids in prepyriform cortex of rats fed imbalanced amino acid diets

Monoamines and amino acids were measured in anterior prepyriform cortex (PPC) and anterior cingulate cortex (CC) of male Sprague-Dawley rats after they were offered basal, imbalanced (IMB) or corrected amino acid diets, limited in threonine (THR) or isoleucine (ILE). In the THR study, brains were taken after 2.5 hr of feeding, when intake of THR-IMB was just depressed. In the ILE study the brains were taken after 3.5 hr on ILE-IMB, a less severely imbalanced ration, before the onset of food intake depression. The PPC has been shown to be involved in the acute response of animals to imbalanced amino acid diets. In the PPC from the IMB diet groups, NE was reduced by 30%, but the other monoamines were unchanged. In CC, an area involved in the adaptive, but not the acute feeding response to imbalanced diets, the monoamines were unchanged in the IMB diet groups. In both studies, in both tissues, the limiting amino acids were decreased in the IMB groups, although the decrease of ILE in the CC failed to reach significance. The remaining indispensable amino acids, added to create the imbalance, were slightly reduced in the THR-IMB group, but not in the ILE-IMB group in both tissues. Thus, the amino acid patterns were altered in the PPC and CC, as they are in whole brains from animals fed imbalanced amino acid diets. These results also suggest that the concentration of NE in the PPC may be associated with the initial food intake response of animals to imbalanced amino acid diets.     

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.     

Parabrachial lesions disrupt responses of rats to amino acid devoid diets, to protein-free diets, but not to high-protein diets

Normal rats "reduce" intake of diets that lack an essential amino acid (THR-DEV), are protein free (PO%), or contain a high proportion of protein (P75%). We tested the importance of the parabrachial nuclei (PBN) in signaling such adjustments of food intake by placing electrophysiologically guided lesions in these nuclei at points that responded to gustatory stimuli. When fed the THR-DEV diet, rats with PBN lesions (PBNx) decreased their food intake significantly less than the controls (78.5 vs. 44.4%). When put on a P0% diet, PBNx animals decreased their intake only 8% compared with 23% for our CONT group. When put on a P75% diet, however, both groups decreased their intake in an equivalent amount. These experiments show that the PBN is involved in the learned aversion to an amino acid devoid diet.     

Effect of amino acid imbalance and deficiency on dietary choice patterns of rats

Detailed dietary choice patterns were determined with a computerized feeding monitoring system in groups of Sprague Dawley rats kept on a 12:12 hr light-dark cycle and offered in sequence a series of dietary choice regimens involving amino acid-imbalanced or deficient diets with threonine as the most limiting amino acid. Animals established their preference for a threonine-basal diet over a threonine-imbalanced or a threonine-devoid (devoid of threonine) diet shortly (within 2-3 hr) after the consumption of small quantities of either diet in the beginning of the first dark-cycle. An intensive sampling process characterized by frequent small bouts was evident throughout the light period. Both the meal size and the meal frequency of the imbalanced or devoid diet were curtailed after prolonged choices. Animals preferred the threonine-corrected (imbalance corrected by threonine supplementation) over the threonine-basal diet initially with an increase in meal frequency. But no clear choice for either diet was observed thereafter. Animals did not establish their preference for the threonine-corrected diet when paired with the threonine-devoid diet until after 5 days with a steady decrease in the meal size of the devoid diet but not the meal frequency. When the protein-free diet was introduced as an alternative for the threonine-imbalanced diet, animals selected the protein-free diet during the first dark-cycle after consuming a small amount of the imbalanced diet. Initially there was a drastic reduction in meal size of the imbalanced diet and subsequently a decrease in meal frequency as well. Nevertheless, animals immediately rejected the protein-free diet and chose the threonine-basal diet when it replaced the imbalanced diet as an alternative. The almost exclusive preference for the basal diet occurred in the beginning of the first dark-cycle with an increase in meal size but no change in meal frequency. The sampling bouts of small quantities, which followed the first introduction of the diets in the choice regimens, may be an inherent investigative behavior whereby the physical or oropharyngeal properties of the diets are recognized. The establishment of the choices for the alternative diets in the present experiments provides additional information about the rapid time course of the food intake control mechanisms in rats fed amino acid-imbalanced or deficient diets.     

Increase in plasma ammonia and amino acids when rats are fed a 44% casein diet

Rats were trained to eat a 6% casein basal diet during a 3-hour period per day. They were then fed either the same 6% casein diet or a 44% casein diet for 3 hours. No food intake depression was observed in the rats eating 44% casein diet during the 3-hour period. Plasma ammonia and amino acids and brain amino acids were measured at 0, 4, 12 and 24 hours after presentation of the 6% or 44% casein diets. Plasma ammonia rose to 134 (p less than 0.01) and 110 micromolar (p less than 0.05) in the 44% casein fed rats at 4 and 12 hours, respectively, as compared to 67 and 53 micromolar, respectively, for the 6% casein fed rats. All plasma amino acid concentrations except methionine and glutamate were elevated (p less than 0.05) at 4 hours. In the brain, threonine, glutamine and tyrosine concentrations were elevated (p less than 0.05) at 4 hours after diet presentation. At 24 hours, valine, isoleucine, leucine, phenylalanine, and methionine concentrations were also elevated (p less than 0.05). Because intake of the 44% casein diet decreases the second day of its presentation, as noted in an earlier experiment, the increases in plasma ammonia and its possible entry into the brain as reflected by increased brain glutamine together with changes in amino acid concentrations should be considered collectively among possible metabolic signals affecting intake of high protein diets.     

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.     

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.     

Effects of age on the feeding response to moderately low dietary protein in rats

Moderately low levels of dietary protein are associated with increased food intake and body fat. We propose that the generation of this feeding signal is dependent on the level of dietary protein relative to the protein requirement of the animal, that is, that protein-dependent feeding is maximized when the level of dietary protein is around the animal's protein requirement. One of the factors that affects an animal's protein requirement is age. We predict that young, growing animals are more responsive to a moderately low level of dietary protein than are mature animals. The feeding response to moderately low dietary protein (10% casein) was determined in young ( approximately 190 g) and more mature ( approximately 340 g) Sprague-Dawley rats for 12 days. As an index of amino acid deamination, serum urea nitrogen concentrations were determined, as was the in vitro release of neuropeptide Y (NPY) from hypothalamic tissue containing the paraventricular nucleus. Young rats were more responsive to the feeding effects of moderately low dietary protein than mature animals. In young rats, cumulative food intake was inversely correlated with serum urea nitrogen concentration. No correlation was found in mature animals. Although the amount of NPY remaining in hypothalamic tissue after incubation was significantly greater (p = 0.04) in young rats fed 10% casein as compared with rats fed the standard 20% casein diet, no dietary difference in K(+)-stimulated NPY release was observed. We hypothesize that the signal for low-protein-induced hyperphagia is a reduction in a compound whose production is coupled to the level of amino acid deamination in the brain.     

Effect of protein intake on weight gain and plasma amino acid levels in uremic rats.

Chronically uremic rats weighing approximately 180-200 g and sham-operated controls of similar weight were pair fed diets containing 5, 15 or 23% protein for 10-12 wk. At each level of protein intake, uremic animals gained less weight and had lower protein efficiency ratios than controls. In addition, certain plasma amino acid levels were altered in the uremic animals. These included tyrosine and the tyrosine/phenylalanine ratio, which were decreased, and citrulline, glycine, and the methylhistidines, which were increased. In both uremic and control rats, plasma concentrations of certain amino acids, primarily nonessential ones, varied inversely with protein intake; with the 5% protein diet, the ratio of essential to nonessential amino acids was significantly reduced. These observations indicate that both uremia and reduced protein intake may affect growth and amino acid metabolism in rats with chronic renal failure. The finding that uremic rats utilize protein less efficiently may indicate that marked reductions in protein intake may be particularly hazardous to the nutritional status of the uremic patient.     

Dietary self-selection and the regulation of protein and energy intake in chicks

Studies were conducted to determine whether chicks could regulate their protein intake independent of total energy intake in self-selection feeding trials. Day-old White Mountain cockerels were reared in electrically-heated battery brooders and given access to either a 23% protein control ration (no choice) or two diets containing 10% or 60% protein with or without supplemental amino acids. The latter were added to either improve the dietary amino acid balance or to alter plasma and brain levels of free large neutral amino acids (tryptophan, isoleucine, leucine, valine, phenylalanine, and tyrosine) which have been implicated in the neuroregulation of feed intake. Both feed and water were provided ad lib, and the location of the feed troughs within each pen were changed daily. Body weights and feed intakes were measured daily, and total calorie and protein intakes were calculated. Chicks offered 10% and 60% protein diets with no supplemental amino acids exhibited reduced weight gains and markedly higher protein intakes as compared to birds fed either the control ration or those given a choice between 10% and 60% protein diets supplemented with methionine. The higher protein consumption by chicks fed the unsupplemented diets most likely was a result of an attempt to compensate for a dietary methionine deficiency. Chicks fed the 10% and 60% protein diets supplemented with amino acids grew at a slower rate than those fed the 23% protein control diet. In general, plasma and brain data did not support a proposed relationship between certain large neutral amino acid ratios and protein or energy intake.     

Dietary sardine protein lowers insulin resistance, leptin and TNF-α and beneficially affects adipose tissue oxidative stress in rats with fructose-induced metabolic syndrome

The present study aims at exploring the effects of sardine protein on insulin resistance, plasma lipid profile, as well as oxidative and inflammatory status in rats with fructose-induced metabolic syndrome. Rats were fed sardine protein (S) or casein (C) diets supplemented or not with high-fructose (HF) for 2 months. Rats fed the HF diets had greater body weight and adiposity and lower food intake as compared to control rats. Increased plasma glucose, insulin, HbA1C, triacylglycerols, free fatty acids and impaired glucose tolerance and insulin resistance was observed in HF-fed rats. Moreover, a decline in adipose tissues antioxidant status and a rise in lipid peroxidation and plasma TNF-α and fibrinogen were noted. Rats fed sardine protein diets exhibited lower food intake and fat mass than those fed casein diets. Sardine protein diets diminished plasma insulin and insulin resistance. Plasma triacylglycerol and free fatty acids were also lower, while those of α-tocopherol, taurine and calcium were enhanced as compared to casein diets. Moreover, S-HF diet significantly decreased plasma glucose and HbA1C. Sardine protein consumption lowered hydroperoxide levels in perirenal and brown adipose tissues. The S-HF diet, as compared to C-HF diet decreased epididymal hydroperoxides. Feeding sardine protein diets decreased brown adipose tissue carbonyls and increased glutathione peroxidase activity. Perirenal and epididymal superoxide dismutase and catalase activities and brown catalase activity were significantly greater in S-HF group than in C-HF group. Sardine protein diets also prevented hyperleptinemia and reduced inflammatory status in comparison with rats fed casein diets. Taken together, these results support the beneficial effect of sardine protein in fructose-induced metabolic syndrome on such variables as hyperglycemia, insulin resistance, hyperlipidemia and oxidative and inflammatory status, suggesting the possible use of sardine protein as a protective strategy against insulin resistance and related situations.     

Induction of conditioned taste aversion in rats by GABA or other amino acids

GABA included in the diet is known to reduce food intake and growth of rats fed a low protein diet. Experiments were designed to determine if GABA or other small neutral amino acids would affect food intake if they were administered separately from the diet, and if such amino acids could induce a conditioned taste aversion (CTA) to saccharin. Intubated or injected GABA or alpha-aminoisobutyric acid (AIB), a non-metabolizable isomer of GABA, reduced food intake. When rats were fed a low protein diet, IP injection of threonine (2 mmoles/200 g rat) induced CTA but did not depress food intake; serine (3 mmoles/200 g rat) induced CTA and caused only a small reduction in food intake. Another isomer, alpha-amino-n-butyric acid did not affect food intake or induce CTA at the tested concentrations. Adaptation to a high protein diet, which increases enzymatic degradation of many amino acids including GABA and serine, lessened severity of GABA-induced CTA and eliminated that caused by serine. CTA to saccharin can be induced by certain amino acids; the mechanism is unknown but may involve malaise or other adverse sensations.     

Amino acid imbalance, a nutritional model: serotonin3 mediation of aversive responses.

The feeding responses to essential dietary amino acid (AA) disproportion have been useful in nutritional studies. These responses involve first: recognition of the imbalance and second: rejection of the diet, likely via development of a learned aversion. In the rat, we have studied the role of the limiting AA and protein synthesis in the recognition phase by replacement of the limiting AA into a brain area essential for the initial depressed feeding response. We have also reported a reciprocal relationship between serotonin (5HT) activity and intake of imbalanced diets. High doses of the 5HT3 receptor antagonist, ICS 205-930 (ICS; 9 mg/kg IP), restored food intake to 85% or more of control intake. In the present experiments, similar treatment with ICS blocked the classical conditioned taste aversion to a saccharin solution paired with lithium chloride. These results suggest that the increased intake of AA imbalanced diets after ICS may be due to 5HT-mediated blockade of a learned aversion.     

Self-selection of dietary casein and soy-protein by the cat

Growing specific-pathogen-free kittens were fed for two weeks a choice between two complete diets differing only in protein content. When casein diets containing 18, 36 and 54% protein were offered in the three possible combinations, the kittens consistently avoided the higher casein diets and kittens offered the two highest levels of casein significantly reduced their total food intake. In one soy-protein choice study, 16, 31 and 63% protein diets were each offered with a protein-free (PF) diet. When diets were similar in physical consistency, kittens selected similar amounts of both diets with the result that the PF:16% group consumed below their requirement of protein. In another soy-protein experiment the 16, 31 and 63% protein diets were offered in their three possible combinations. Kittens in all three groups selected similar amounts of both diets. Except for their avoidance of casein, the kittens did not regulate in a consistent manner their intake of protein and therefore, behaved very differently from the rat in the self-selection of dietary protein.     

Avoidance of GABA-containing diets by olfactory bulbectomized rats

In order to test the importance of olfaction in the avoidance of GABA-containing diets by intact rats, olfactory bulbectomized and sham-operated control rats were allowed to choose between a low protein control diet and this diet supplemented with 2.5% GABA; the specificity of the response was examined by also testing for responses to other amino acids. Both groups of rats markedly avoided the GABA diet initially; the bulbectomized rats later tended to increase their intake of this diet. The bulbectomized rats chose similar amounts from the control diet and one supplemented with alanine whereas the control rats ultimately strongly preferred the latter diet. Both groups only moderately avoided a threonine-supplemented diet. The results suggest that (1) the odor of GABA is not critical in the avoidance of diets containing this amino acid; and (2) patterns of food selection from diets containing GABA differ from choices from diets containing other small neutral amino acids such as alanine or threonine.     

Influence of taste on dietary choice of rats fed amino acid imbalanced or deficient diets

Diets with added quinine as the negative taste cue or saccharin as the positive taste cue were employed to determine the influence of taste on dietary choice of rats offered diets containing different proportions of amino acids (amino acid imbalance) and which differed in acceptability. The quinine was added to the protein-free or the corrected (corrected for amino acid imbalance) diet that animals normally preferred and the saccharine was added to the amino acid imbalanced or deficient diets that animals normally avoided. There appeared to be a balance in acceptance of a diet between the undesirability of the quinine and the degree of metabolic benefit from the diets with favorable metabolic or nutritional characteristics. Although the presence of higher levels of quinine could interfere with the normal dietary preference based on metabolic consequences, the animals invariably selected the metabolically favorable diet if they were forced to experience the metabolic characteristics of the diets by having to consume them exclusively. The presence of the taste cues appeared to enhance the acceptance or avoidance of diets in the choice regimens, possibly by aiding in their identification, especially to animals previously not exposed to the taste cues.     

Dietary amino acid imbalance and neurochemical changes in three hypothalamic areas

The impact of feeding imbalanced amino acid diets on monoamine, metabolite and amino acid concentrations was measured in the ventromedial hypothalamus (VMH), lateral hypothalamus (LH) and paraventricular nucleus (PVN). After rats were fed either an isoleucine imbalanced diet, a threonine imbalanced diet, or the appropriate basal or corrected control diets, regional differences were found in neurochemical concentrations. Contrary to our expectations, the limiting amino acid was unchanged in the imbalanced groups, tending to be decreased only in the isoleucine imbalanced-diet group in the PVN. This is the first report that the limiting amino acid was not reduced uniformly in the brain after imbalanced amino acid feeding. In the VMH, norepinephrine (NE) was increased by 22% and 63% in the threonine and isoleucine imbalanced-diet groups, respectively. Since the concentration of NE was affected even before the decrease in feeding, both in the VMH, and, as previously reported, in the prepyriform cortex, the NE system may be involved in very early responses to imbalanced amino acid diets.     

Development of renal lesions in dogs after 11/12 reduction of renal mass. Influences of dietary protein intake

Renal failure was induced in 15 normal Beagle dogs by ligation of approximately 5/6 of the renal arteries of the left kidney and contralateral nephrectomy in order to determine how: (a) 11/12 reduction in total renal mass influences urine protein excretion and renal morphology in dogs, and (b) dietary protein intake influences renal function, urine protein excretion, and renal morphology in canine renal failure. Dogs were fed a reduced protein diet for 12 weeks after induction of renal failure, while compensatory renal hypertrophy developed. Renal function was then evaluated and dogs were distributed into 2 groups with approximately equal degrees of renal dysfunction. One group was fed a high protein diet (42% protein) and a second group was fed moderately restricted protein diets (18% protein). After 8 weeks, renal function, magnitude of proteinuria, and renal morphology were re-evaluated. Inulin clearance increased in all dogs fed the 42% protein diet and 3 of 10 dogs fed the 18% protein diets. Proteinuria was significantly greater in dogs fed the high protein diet than dogs fed the reduced protein diets. Compared with previously nephrectomized contralateral control kidneys, glomerular sclerosis and renal interstitial lesions had developed in all dogs, regardless of severity of renal dysfunction or diet fed. Although reduced dietary protein intake did not prevent development of renal lesions, renal lesions were significantly more severe in the 5 dogs fed the 42% protein diet and 3 dogs fed the 18% protein diets in which inulin clearance increased, than in 7 dogs fed the reduced protein diets in which inulin clearance did not increase.