Intravenous administration of oxytocin in rats acutely decreases deprivation-induced chow intake,but it fails to affect consumption of palatable solutions

Despite its limited ability to cross the blood-brain barrier, peripherally administered oxytocin (OT) acutely decreases food intake, most likely via the brainstem and hypothalamic mechanisms. Studies performed to date have focused mainly on the effects of subcutaneous or intraperitoneal OT on the consumption of only solid calorie-dense diets (either standard or high-fat), whereas it is unknown whether, similarly to central OT, peripherally administered peptide reduces intake of calorie-dilute and non-caloric palatable solutions. In this project, we established that 0.1 μg/kg intravenous (IV) OT is the lowest anorexigenic dose, decreasing deprivation-induced standard chow intake by ca. 40% in rats and its effect does not stem from aversion. We then used this dose in paradigms in which effects of centrally acting OT ligands on consumption of palatable solutions had been previously reported. We found that IV OT did not change episodic intake of individually presented palatable solutions containing 10% sucrose, 0.1% saccharin, combined 10% sucrose-0.1% saccharin or 4.1%. Intralipid and it failed to affect daily scheduled consumption of a sucrose solution in non-deprived rats. In a two-bottle choice test, IV OT did not shift animals’ preference from sucrose to Intralipid. Finally, OT injected IV prior to the simultaneous presentation chow and a sucrose solution in food-deprived rats significantly decreased chow intake, whereas sugar water consumption remained unchanged. We conclude that IV OT reduces deprivation-induced chow intake without causing aversion, but the dose effective in decreasing energy-driven consumption of high-calorie food fails to affect consumption of palatable calorie-dilute solutions.     

Influence of Feeding Paradigm in Rats on Temporal Pattern of: I. Macronutrient Intake and Arterio-Venous Differences in Plasma Glucose, Insulin and Tryptophan

Relationships among feeding paradigm (single diet vs food selection) and arterio-venous differences (δAV) of glucose, insulin and tryptophan were studied by measuring the temporal patterns of food intake and plasma parameters during 8 hr feeding cycles in rats. Adult male Sprague-Dawley rats were offered a single diet of fixed composition (20% casein) or a choice between two isocaloric diets (0% and 60% casein) for 2 weeks under 8-hr daily feeding conditions, food being offered during the dark cycle. Groups of animals were then killed at the beginning and at 2-hourly intervals throughout the feeding period. With both feeding paradigms, rats showed temporal patterns of energy, carbohydrate and protein intakes with a peak at the beginning and a trough at the end of the feeding period. However, in rats offered a dietary choice the intake of carbohydrate was significantly lower, and the intakes of energy and protein significantly higher than those found in rats offered a single diet. Throughout the feeding period, these differences between single and choice diets became less accentuated in the case of carbohydrate intake, but more accentuated for energy and protein intakes. Paradoxically, rats fed a choice of diets had a significantly lower weight gain than rats fed a single diet. The temporal variation of insulin secretion and tryptophan absorption varied inversely with the two diet paradigms. Moreover, in rats offered a choice of diets, macronutrient intake was significantly correlated with insulin secretion and venous glucose concentration. The opposed physiologic and metabolic responses to the feeding paradigms suggest the need for future studies to examine the possibility that such can function as synchronizers of biological rhythms.     

Influence of Feeding Paradigm in Rats on Temporal Pattern of: I. Macronutrient Intake and Arterio-Venous Differences in Plasma Glucose,Insulin and Tryptophan

Relationships among feeding paradigm (single diet vs food selection) and arterio-venous differences (δAV) of glucose, insulin and tryptophan were studied by measuring the temporal patterns of food intake and plasma parameters during 8 hr feeding cycles in rats. Adult male Sprague-Dawley rats were offered a single diet of fixed composition (20% casein) or a choice between two isocaloric diets (0% and 60% casein) for 2 weeks under 8-hr daily feeding conditions, food being offered during the dark cycle. Groups of animals were then killed at the beginning and at 2-hourly intervals throughout the feeding period. With both feeding paradigms, rats showed temporal patterns of energy, carbohydrate and protein intakes with a peak at the beginning and a trough at the end of the feeding period. However, in rats offered a dietary choice the intake of carbohydrate was significantly lower, and the intakes of energy and protein significantly higher than those found in rats offered a single diet. Throughout the feeding period, these differences between single and choice diets became less accentuated in the case of carbohydrate intake, but more accentuated for energy and protein intakes. Paradoxically, rats fed a choice of diets had a significantly lower weight gain than rats fed a single diet. The temporal variation of insulin secretion and tryptophan absorption varied inversely with the two diet paradigms. Moreover, in rats offered a choice of diets, macronutrient intake was significantly correlated with insulin secretion and venous glucose concentration. The opposed physiologic and metabolic responses to the feeding paradigms suggest the need for future studies to examine the possibility that such can function as synchronizers of biological rhythms.     

Perception of sweet taste is important for voluntary alcohol consumption in mice

To directly evaluate the association between taste perception and alcohol intake, we used three different mutant mice, each lacking a gene expressed in taste buds and critical to taste transduction: α-gustducin ( Gnat3 ), Tas1r3 or Trpm5 . Null mutant mice lacking any of these three genes showed lower preference score for alcohol and consumed less alcohol in a two-bottle choice test, as compared with wild-type littermates. These null mice also showed lower preference score for saccharin solutions than did wild-type littermates. In contrast, avoidance of quinine solutions was less in Gnat3 or Trpm5 knockout mice than in wild-type mice, whereas Tas1r3 null mice were not different from wild type in their response to quinine solutions. There were no differences in null vs. wild-type mice in their consumption of sodium chloride solutions. To determine the cause for reduction of ethanol intake, we studied other ethanol-induced behaviors known to be related to alcohol consumption. There were no differences between null and wild-type mice in ethanol-induced loss of righting reflex, severity of acute ethanol withdrawal or conditioned place preference for ethanol. Weaker conditioned taste aversion (CTA) to alcohol in null mice may have been caused by weaker rewarding value of the conditioned stimulus (saccharin). When saccharin was replaced by sodium chloride, no differences in CTA to alcohol between knockout and wild-type mice were seen. Thus, deletion of any one of three different genes involved in detection of sweet taste leads to a substantial reduction of alcohol intake without any changes in pharmacological actions of ethanol.     

Regulation of Amino Acid Intake in the Rat: Lysine Preference with Gluten Diets of Varying Lysine Content

Weanling rats were offered a choice of two diets, varying only in lysine content. The preference test was performed between a 20% gluten diet supplemented with 0.5% lysine HCl and a lysine-deficient amino acid mixture, 20% gluten or 20% gluten plus 0.2% lysine HCl diet, and was done between a 20% gluten diet and a 20% gluten diet containing 3, 5 or 7% lysine · HCl. Weight gain and food intake were not significantly different among all the self-selecting groups, and these values were almost the same as those in rats fed only the 20% gluten supplemented 0.5% lysine · HCl diet which obtained maximal growth. The lysine intake of the combination groups ranged from 67 to 240 mg per day (0.62-2.35% of consumed food). It was demonstrated that rats exhibit a definite ability to regulate lysine intake, and they select sufficient lysine to meet their requirements for this amino acid. The self-selection technique may be useful as a method to determine the requirements for amino acids.     

Striatal-Enriched Protein Tyrosine Phosphatase Controls Responses to Aversive Stimuli: Implication for Ethanol Drinking

The STriatal-Enriched protein tyrosine Phosphatase (STEP) is a brain-specific phosphatase whose dysregulation in expression and/or activity is associated with several neuropsychiatric disorders. We recently showed that long-term excessive consumption of ethanol induces a sustained inhibition of STEP activity in the dorsomedial striatum (DMS) of mice. We further showed that down-regulation of STEP expression in the DMS, and not in the adjacent dorsolateral striatum, increases ethanol intake, suggesting that the inactivation of STEP in the DMS contributes to the development of ethanol drinking behaviors. Here, we compared the consequence of global deletion of the STEP gene on voluntary ethanol intake to the consumption of an appetitive rewarding substance (saccharin) or an aversive solution (quinine or denatonium). Whereas saccharin intake was similar in STEP knockout (KO) and wild type (WT) littermate mice, the consumption of ethanol as well as quinine and denatonium was increased in STEP KO mice. These results suggested that the aversive taste of these substances was masked upon deletion of the STEP gene. We therefore hypothesized that STEP contributes to the physiological avoidance towards aversive stimuli. To further test this hypothesis, we measured the responses of STEP KO and WT mice to lithium-induced conditioned place aversion (CPA) and found that whereas WT mice developed lithium place aversion, STEP KO mice did not. In contrast, conditioned place preference (CPP) to ethanol was similar in both genotypes. Together, our results indicate that STEP contributes, at least in part, to the protection against the ingestion of aversive agents.     

Rapid calorie metering inad lib rats

Adult wistar rats of either sex housed in individual cages and fedad lib were used to investigate the effects of calorie content and taste on ingestion. A 15 min single-choice solution (water, quinine, sucrose, saccharin) test as well as 1 h mixed diet (stock diet, sucrose-mixed, saccharin-mixed, quinine-mixed) tests were used. Calorically-rich sucrose either in solution or in mixed-diet was preferentially ingested. Tasteper se has not influenced calorie intake, as intake on sweet saccharin (3.4 ±0.4 cal) or on bitter quinine (3.3 ±0.6 cal) was similar to intake on stock diet (4.8 ±0.8 cal). Increased 5 min intake on sucrose solution (4.6 ±0.1 ml) over intake of other test solutions (saccharin 3.5 ±0.2 ml, quinine, 1.2 ±0.3 ml, water 2.1 ±0.1 ml) and calorie intake suppression on 1 h stock diet immediately following sucrose solution intake, indicate rapid calorie metering, probably based on fast-acting specific gustatory signals     

Food preference of young rhesus monkeys fed marginally zinc deficient diets

Zinc deprivation has been shown to produce hypogeusia in rhesus monkeys but the possible consequences of altered taste sensitivity for food preference and selection is not known. We studied 13 rhesus monkeys (2.5 to 3 years of age) for food preference in a structured choice situation. These animals were fed two levels of dietary zinc: marginally zinc deprived (fed 4 μg zinc/g diet from conception and throughout development;N=6), or control (fed a 100 μg zinc/g diet throughout the study;N=7). Preference for familiar vs unfamiliar food items, order of food preferences, persistence in retrieval of preferred and nonpreferred foods, and preference under four deprivation periods were examined. Animals were offered a choice of two food items, both containing minimal zinc levels, and food choice and latency of choice were measured. Results indicate that animals fed the marginal zinc diets had reduced preference for unfamiliar foods relative to controls and different patterns of food preference. These results may be relevant to maintenance of appropriate food selection in marginally zinc deprived populations.     

A trial of assessment of the interrelated action of metabolic and sensory factors in the choice of food by rats in various states of protein and calcium nutrition

The strategy of food choice by adult rats was studied after giving previously to them a balanced diet, a non-protein diet and/or a diet with calcium excess. Tests were carried out for food preference offering to the rats semi-purified diets differing in the contents of protein and calcium. Besides that, the taste of the food was changed adding to it saccharose, quinine, mint and caramel. Depending on the nutritional state of the rats significant differences were found in their preference for foods which was related to the general direction of homeostasis recovery. In protein malnutrition diets were chosen containing protein, while calcium content and taste were of secondary importance.     

Glossopharyngeal nerve transection does not alter taste reactivity to sucrose conditioned to be aversive

Glossopharyngeal nerve (GL) transection in rats is known to markedly reduce gaping, a stereotypical aversive oromotor behavior, in response to intraorally delivered quinine. In this experiment we tested whether GL transection would reduce gaping in response to an otherwise palatable stimulus (sucrose) but conditioned to be aversive. Sprague-Dawley rats were implanted with intraoral cannulae. Five received bilateral transection of the GL and five served as sham-operated controls. Water-deprived rats were presented with 0.3 M sucrose for 15 min immediately followed by an injection of 0.15 M LiCl on three occasions. Rats were then habituated to the taste reactivity chamber and intraoral fluid infusion for 3 days, and tested on day 4 with a 1 ml infusion (1 min) of 0.3 M sucrose. All rats drank negligible amounts of sucrose by the third conditioning session and there were no differences in sucrose intake between the groups. There were no significant differences in gapes, or any other measured oromotor response, to sucrose between GL-transected and sham-operated rats. These results show that the GL is not a necessary afferent limb for gaping in response to conditionally aversive taste compounds.     

Acute 3rd-ventricular amylin infusion potently reduces food intake but does not produce aversive consequences

In this study, a conditioned taste aversion (CTA) paradigm was used to assess the possibility that 3rd-ventricular (i3vt) administration of the pancreatic hormone amylin produces aversive consequences that secondarily reduce food intake independently of the normal regulation of energy balance. After 1-h daily access to water for 7 days, rats were given 1-h access to a 0.15% saccharin solution, followed immediately by i3vt amylin (100 pmol) in one group (n=7) and i3vt CSF vehicle in another (n=7). As positive control for the formation of a CTA, a third group of seven rats received intraperitonial (i.p.) lithium chloride (LiCl). Saline was given i.p. to a fourth group (n=7) as control for i.p. LiCl. As expected, the LiCl rats exhibited a marked aversion to the saccharin in a subsequent two-bottle intake test. In contrast, although the 100 pmol i3vt amylin dose is substantially higher than that required to reduce food intake, no evidence of a CTA was observed in the rats that had received i3vt amylin. In summary, these data are consistent with the conclusion that acute i3vt amylin infusion does not reduce food intake by producing aversive consequences.     

Diet switching in a generalist mammalian folivore: fundamental to maximising intake

Generalist mammalian herbivores exploit a diverse diet. A generalised feeding strategy utilises a mixed diet to obtain a range of nutrients and to reduce the detoxication load of similar groups of plant secondary metabolites (PSMs). There is limited research investigating how mammalian herbivores achieve this dietary mixing in their daily foraging activities. We investigated the patterns of, and behaviours associated with, dietary mixing in a generalist mammalian folivore, the common brushtail possum (Trichosurus vulpecula). Possums were offered foliage of two eucalypt species (Eucalyptus globulus and E. regnans) as either (a) Full choice: both species offered for 8 h; (b) Restricted choice: both species offered for 2×2 h blocks; (c) G–R no choice: E. globulus offered for the first 4 h, E. regnans offered for next 4 h; and (d) R–G no choice: E. regnans offered for first 4 h, E. globulus offered for next 4 h. We hypothesised that possums would maximise intake on the Full choice diet, where time availability was greatest in combination with a choice of foliage. We also hypothesised that diet switching, defined as the frequency of changing between food types while feeding, would play a fundamental role in maximising intake. Possums achieved maximum intake on the Full choice diet and minimum intake on the No choice diets. Although intake was similar between the Restricted choice and No choice diets, possums achieved this intake in half the amount of time when able to switch regularly between foliage on the Restricted choice diet. We conclude that a generalist herbivore’s ability to effectively switch diets when foraging is fundamental to maximising intake. Hence, the degree of plant heterogeneity in an environment, and the spatial scale at which it occurs, may affect an herbivore’s foraging decisions and, ultimately, influence its foraging efficiency.     

Relationships between taste reactivity and intake in the neurologically intact rat

To investigate the differences between short-term intake testsand taste reactivity responses to tastes, rats received 1-minintraoral infusions of a variety of tastants delivered at therate of 1 ml/min. Oral responses were videotaped and analysedin terms of the sequence and number of ingestive and aversivetaste reactivity response components evoked. Intake was alsomeasured. The number of rats displaying ingestive taste reactivitycomponents and the mean number of ingestive components displayedper rat elicited by sucrose and NaCl increased with increasingconcentration. Intake was high across all concentrations. HClinfusions elicited alternation between ingestive and aversiveresponse components. The number of rats displaying aversivetaste reactivity response components and the mean number ofaversive response components displayed per rat, elicited byQHCl, increased with increasing concentration, while both intakeand the median latency to reject QHCl decreased (ExperimentI). To determine whether other tastes judged bitter by humanswould elicit a quinine-like taste reactivity response in therat, sucrose octa-acetate (SOA), quinine sulfate (QS) and caffeine(CAF) stimuli were examined. Both QS and CAF infusions elicitedan increased number of aversive response components with increasingconcentration, and intake decreased. SOA infusions elicitedalternation between ingestive and aversive response componentsfollowed by a display of solely aversive components, and bothintake and median latency to reject the infusions decreasedsignificantly with increased concentration (Experiment II).Experiment I demonstrated that hypertonic NaCl infusions elicitingestive response components, while short-term intake testsshow that hypertonic NaCl is rejected and is thus inferred tobe aversive. Rats received prolonged infusions of hypertonicNaCl solutions at the rate of 1 ml/min until fluid was seenon the surface of the chamber, indicating rejection. Prolongedinfusions of hypertonic NaCl solutions elicited an initial displayof solely ingestive response components followed by an abruptshift to solely aversive response components and active fluidrejection. Higher concentrations elicited this shift soonerthan lower ones (Experiment III). The results suggest that patternsof taste reactivity response components are good predictorsof intake duration.     

Selective decrease in protein intake following brain serotonin depletion.

The effect of reduced brain serotonin concentration achieved with drugs or raphé lesions on the control of protein intake and energy intake by young rats was examined. All rats were provided an oppurtunity to select from 15% and 55% casein diets provided simultaneously in two food cups. Systemic parachlorphenylalanine, central 5, 7-dihydroxytryptamine or mid-brain raphé lesions reduced by 20–30% the amount of protein consumed from these two diets over a two week study period. Across these same groups total energy intake was not different from the control groups. This decrease in the proportion of dietary energy selected as protein by the treated self-selecting rats was associated with the reduction observed in brain serotonin and its major metabolite, 5-hydroxyindole acetic acid at the completion of the feeding period. These findings support previous work implicating brain serotonin metabolism in the selective control of protein intake.     

Re-examination of the poisoned-partner effect with the two-bottle testing method

Revusky, Coombes and Pohl (Animal Learning and Behavior 10 (1982) 83–90) suggested that poisoned conspecifics function as aversive unconditional stimuli (USs) for rats (the poisoned-partner effect). The present study re-examines this effect with the two-bottle testing method. Rats were first given saccharin and later exposed to a poisoned conspecific. They exhibited an aversion to saccharin in the subsequent tests involving a choice between saccharin solution and water. Furthermore, the subjects that were only exposed to the poisoned conspecific later avoided saccharin. The poisoned-partner effect, therefore, can also be explained by sensitization. This finding suggests that poisoned conspecifics are simply salient stimuli that attract the rat’s attention, rather than aversive USs of food aversion learning for rats.     

Centrally Administered Ghrelin Acutely Influences Food Choice in Rodents

We sought to determine whether the orexigenic hormone, ghrelin, is involved in the intrinsic regulation of food choice in rats. Ghrelin would seem suited to serve such a role given that it signals hunger information from the stomach to brain areas important for feeding control, including the hypothalamus and reward system (e.g. ventral tegmental area, VTA). Thus, in rats offered a choice of palatable foods (sucrose pellets and lard) superimposed on regular chow for 2 weeks, we explored whether acute central delivery of ghrelin (intracerebroventricular (ICV) or intra-VTA) is able to redirect their dietary choice. The major unexpected finding is that, in rats with high baseline lard intake, acute ICV ghrelin injection increased their chow intake over 3-fold, relative to vehicle-injected controls, measured at both 3 hr and 6 hr after injection. Similar effects were observed when ghrelin was delivered to the VTA, thereby identifying the VTA as a likely contributing neurobiological substrate for these effects. We also explored food choice after an overnight fast, when endogenous ghrelin levels are elevated, and found similar effects of dietary choice to those described for ghrelin. These effects of fasting on food choice were suppressed in models of suppressed ghrelin signaling (i.e. peripheral injection of a ghrelin receptor antagonist to rats and ghrelin receptor (GHSR) knock-out mice), implicating a role for endogenous ghrelin in the changes in food choice that occur after an overnight fast. Thus, in line with its role as a gut-brain hunger hormone, ghrelin appears to be able to acutely alter food choice, with notable effects to promote “healthy” chow intake, and identify the VTA as a likely contributing neurobiological substrate for these effects.     

Central mechanisms of roles of taste in reward and eating

Taste is unique among sensory systems in its innate association with mechanisms of reward and aversion in addition to its recognition of quality, e.g., sucrose is sweet and preferable, and quinine is bitter and aversive. Taste information is sent to the reward system and feeding center via the prefrontal cortices such as the mediodorsal and ventrolateral prefrontal cortices in rodents and the orbitofrontal cortex in primates. The amygdala, which receives taste inputs, also influences reward and feeding. In terms of neuroactive substances, palatability is closely related to benzodiazepine derivatives and beta-endorphin, both of which facilitate consumption of food and fluid. The reward system contains the ventral tegmental area, nucleus accumbens and ventral pallidum and finally sends information to the lateral hypothalamic area, the feeding center. The dopaminergic system originating from the ventral tegmental area mediates the motivation to consume palatable food. The actual ingestive behavior is promoted by the orexigenic neuropeptides from the hypothalamus. Even palatable food can become aversive and avoided as a consequence of a postingestional unpleasant experience such as malaise. The neural mechanisms of this conditioned taste aversion will also be elucidated.     

Diazepam-binding inhibitor-like activity in rat cerebrospinal fluid after stimulation by an aversive quinine taste

Cerebrospinal fluid (CSF) taken from rats after stimulation by an aversive quinine taste (hereafter called quinine CSF) administered into the fourth ventricle of mice suppressed their intake of 5% sucrose solution. We examined the effects of CSF on glutathione-induced tentacle ball formation (TBF) of hydra to determine the change in CSF components associated with aversive taste stimuli. The suppressive activity of quinine CSF on TBF in the presence of 3 microM S:-methyl-glutathione (GSM) was markedly lower than that of CSF obtained from control rats (control CSF). Pronase-treated quinine CSF had suppressive activity similar to that of control CSF. The active principle passed through an ultrafiltration membrane, with a molecular weight cut-off of 30 kDa, but not through one with a cut-off of 3 kDa. A peptide fragment of diazepam-binding inhibitor (DBI) nullified the suppression of TBF at 3 microM GSM by control CSF. The nullifying activity of quinine CSF was not observed after treatment with a benzodiazepine receptor preparation that was able to bind DBI. When flumazenil, a benzodiazepine receptor antagonist, was given to mice, the suppression of the intake of 5% sucrose solution by quinine CSF was partially reversed. It is suggested that quinine CSF contains a DBI-like substance.     

Effect of restricted feed intake and addition of vitamins B2 and B6 and folic acid on the liver concentration of zinc and copper in rats

The aim of the present study was to investigate the influence of nutritional deficiency and dietary addition of vitamins (B₂, B₆, and folate) on hepatic concentration of zinc and copper in rats. The experiment was performed on 260 growing male Wistar rats divided into 13 groups. Animals of 11 groups were fed isocaloric diets (14.7 MJ/kg) in which the 20% of energy was derived from protein. Another two groups of rats were offered diets with 9% or 4.5% of energy originating from protein. Animals of both mentioned groups and of the control group (20% of energy from protein) were offered diets ad libitum. The other 10 groups were offered 50% and 30% of the amount consumed in the control group. Eight groups, from those 10 restricted ones, were differentiated by dietary addition of vitamins B₂ and B₆ and folate (300% addition). Restricted feed intake did not affect the liver zinc concentration but significantly increased the copper concentration. The addition of vitamin B₆ decreased the liver Zn concentration. The highest liver Cu concentration was noted in rats offered restricted diets to only 30% of intake in the control group and high in vitamin B₂ and in rats supplemented with all of studied vitamins together. It suggests that vitamin B₂ had the strongest impact on liver Cu concentration in rats fed restricted diets.     

Learned aversions and taste qualities in hamsters

Interralations among taste perceptions in gloden hamsters (Mesocricetusauratus) were examined using generalizaions of learned tasteaversions. If stimulus A is avoided given a taste aversion hasbeen established to stimulus B, and vice versa, A and B ‘cross-generalize’.Stimuli within five groups cross-generalized. The groups ofcompounds were (i) sweeteners (fructose, saccharin, sucrose);(ii) sodium salts (NaCl, NaNO₃, Na₂SO₄): (iii) non-sodium salts(KCl, MgSO₄ NH₄Cl) plus quinine HCl; (iv) acids (acetic, hydrochloric,citric); and (v) urea. Only two pairs of stimuli from differentgroups cross-generalized (HCl—NH₄Cl. quinine HCl—urea).Neural patterns of response recorded form chorda tympam nervefibers in hamsters suggest that taste receptors on the anteriortongue distinguish among three groups of taste stimuli: sweeteners,sodium salts, and a group including non-sodium salts, acids,quinine HCl and urea. Neurons innervating other taste fieldsare likely to provide the information that hamsters use to discriminateamong the tastes of non-sodium-salt and non-sweetener stimuli.