Cholecystokinin, amphetamine and diazepam and feeding in lean and obese Zucker rats.

The hyperphagia characteristic of some types of obesity may result from a deficiency in one or more components of the systems controlling satiety which in rats may include the gastrointestinal hormone cholecystokinin (CCK). Obesity may also influence responsivity to often used central nervous system (CNS)-acting drugs and combination of drugs. In these experiments it was shown that: (1) Zucker fatty rats were less sensitive than lean to intraperitoneal injections of 20 U/kg CCK after a 6-hr fast and when reduced were less sensitive than lean and less sensitive than when obese to injections of 5 U/kg CCK; (2) Although fatties were equally sensitive as leans to injections of 0.5 and 1.0 mg/kg d-amphetamine sulfate, when reduced, they were less sensitive; (3) Injections of 1.25 and 2.5 mg/kg diazepam produced smaller increases in food intake after a 6-hr fast in fatty and reduced fatty than lean rats; (4) Combination of diazepam with cholecystokinin in both fatty and lean rats produced feeding similar to that following injection of carrier; and (5) A similar additive effect was obtained in both fatty and lean rats when diazepam was combined with amphetamine; however, the fatty appeared to be more sensitive to the amphetamine than the diazepam effect. Thus the Zucker fatty rat appears to be less sensitive to these chemicals which affect food intake, which supports the contention that their CNS is generally less responsive.     

NPY presynaptic actions are reduced in the hypothalamic mpPVN of obese (fa/fa), but not lean, Zucker rats in vitro

1. Neuropeptide Y (NPY) profoundly enhances feeding when injected intracerebroventricularly, or directly into hypothalamic nuclei, such as the paraventricular nucleus (PVN). Paradoxically, NPY has a reduced action on feeding in obese Zucker rats relative to lean Zucker rats, although the obese rats have much higher levels of hypothalamic NPY expression. GABAergic inputs to a subpopulation of medial parvocellular PVN (mpPVN) neurons are sensitive to NPY. Here, we tested the hypothesis that the blunted eating response to NPY observed in obese Zucker rats will be reflected in a reduced NPY action at mpPVN GABAergic synapses. 2. 'Blind' whole-cell patch-clamp recordings made from mpPVN neurons in acute brain slices of lean and obese Zucker rats revealed GABAergic inhibitory postsynaptic currents (IPSC) responses which were inhibited by NPY. While the maximum response in the obese Zucker rats was significantly less than in lean Zucker or Sprague-Dawley rats, there was no difference in the EC(50). 3.Experiments using blocking concentrations of Y(1)- or Y(5)-receptor antagonists revealed no differences between lean and obese Zucker rats in the contributions of either of these receptors to the total NPY response in mpPVN. 4. NPY is less effective at the mpPVN GABA synapse in obese than in lean Zucker rats. This is not associated with a change in the proportion of Y(1) or Y(5) receptors mediating the NPY response, and is consistent with the downregulation of NPY receptors or a reduction in receptor-effector coupling, and with the reduced sensitivity of obese rats to NPY.     

Naloxone suppresses feeding and drinking but not wheel running in rats

The effects of naloxone hydrochloride on food and water intake and number of wheel revolutions were measured in male rats. The administration of 10 mg/kg naloxone but not 1 mg/kg suppressed the 3-hr food and water intake in nondeprived rats. Naloxone injections (1 mg/kg or 10 mg/kg) were ineffective in altering the number of wheel revolutions in nondeprived or food deprived rats. These results support the interpretation that the suppressive effects of naloxone previously reported with deprived rats are evident in nondeprived rats and are specific to feeding and drinking.     

Enhanced sensitivity to anorexia and consumption of drinking water induced by interleukin-1 beta in obese yellow mice.

Exogenously administered interleukin-1 beta (IL-1) was reported to suppress food intake and endogenous CRF in the brain was reported to be involved in mediating the IL-1-induced anorexia. The present study was undertaken to investigate the effect of IL-1 (i.p.) on food and water intake in obese yellow mice and in lean mice. Enhanced sensitivity to IL-1-induced suppression of food and water intake were observed in obese yellow mice compared to lean mice (food intake suppression: lean 29.92%, obese 88.48%; water intake suppression: lean 25.18%, obese 71.56% of respective controls). After treatment of the mice with ibuprofen (10 mg/kg i.p.), the suppression of food and water intake was prevented in lean mice but unaffected in obese mice. The results suggest that there may be differential sensitivity to activation of CRF neurons induced by the peripheral injection of IL-1 in obese yellow and in lean mice.     

Significance of endothelin on volume homeostasis in obese Zucker rats

1. The present study explored the impact of endothelin (ET) and its interaction with renal sympathetic nerves in the control of volume homeostasis in obesity. 2. Groups of lean and obese Zucker rats with innervated and denervated kidneys were subjected to an acute isotonic saline volume expansion (VE), 10% bodyweight and renal haemodynamics and excretory function were evaluated following administration of SB209670 (a non-selective ET antagonist) or UK 350 926 (a selective ETA receptor antagonist). 3. Volume expansion in untreated obese rats resulted in a blunted cumulative urine sodium excretion (CuUNaV) after 40 min, VE compared with untreated lean rats being 36 and 51% in the denervated and innervated kidneys, respectively (both P < 0.001). 4. In lean rats, both SB209670 and UK 350 926 caused a depressed ability to excrete the saline load and CuUNaV after 40 min, VE compared with untreated being decreased by 51 and 60% in the denervated and innervated kidneys, respectively (both P < 0.001). 5. SB209670 and UK 350 926 given to obese rats reduced (both P < 0.001) CuUNaV after VE by 43% in denervated kidneys compared with untreated obese rats, whereas they were without effect on the magnitude of the excretory response in innervated kidneys. 6. These findings show that the blunted renal excretory responses to VE present in obese rats were not mediated by ET. Conversely, ET, acting through ETA receptors, plays a role as a diuretic and natriuretic factor in maintaining volume homeostasis by, at least in lean rats, renal nerve-independent mechanisms.     

Abnormal sympatho-adrenal function and plasma catecholamines in obese Zucker rats

The functional integrity of the peripheral sympathetic nervous system and adrenal medulla was assessed in homozygous, lean and obese, 7–8 month old male Zucker rats by the changes in plasma catecholamines during cold and immobilization stresses. Five of eight obese, but no lean rats died during a 24 hr cold stress (4–7°C) from hypothermia. While both lean and obese rats had decreased rectal temperatures after 4 hr of cold stress, the obese had lower temperatures, relatively less of an increase of plasma norepinephrine (NE) and epinephrine (E) than the lean rats, and were unable to consistently maintain their temperatures even during intravenous NE infusions. Obese rats had lower rectal temperatures and higher plasma NE and dopamine levels at 21–22°C ambient temperature, a relative failure to increase plasma NE and E levels after 1 hr of immobilization, but normal or supranormal plasma catecholamine levels after decapitation compared to the lean rats. These results suggest that the obese Zucker rat has abnormalities of both peripheral sympatho-adrenal function and thermoregulation, which may play roles in the development and/or maintenance of many of the physiological and metabolic defects in this animal model of genetic obesity.     

Enhanced sensitivity to naltrexone-induced drinking suppression of fluid intake and sucrose consumption in maternally separated rats

Early-life stress has been identified as a risk factor in the development of a host of disorders, including substance abuse; however the link between early postnatal stress and changes in measures of reward has not been thoroughly researched. The current study had two main objectives: 1) to determine the impact of maternal separation (an animal model of early-life stress) on the consumption of 10% and 2.5% sucrose solutions by Long-Evans rat dams and male and female offspring, and 2) to determine the effect of the opioid antagonist naltrexone (0.1-3.0 mg/kg) on drinking by each of those groups. Dam-pup separations occurred for varying lengths of time during the first two postnatal weeks. In Experiment 1, a two-bottle choice test (sucrose solution vs. water) was administered across five days to both nonhandled (NH) and maternally-separated (MS) offspring as adults and to dams 2-4 weeks post-weaning. In Experiment 2, naltrexone was administered prior to two-bottle choice tests. MS males and the dams of MS litters exhibited increased intake of total fluid and sucrose solutions, whereas results from females were less consistent. Naltrexone elicited a greater decrease in fluid intake and sucrose intake in male MS offspring compared to male NH offspring. These results indicate that early postnatal stress alters both sucrose consumption, a non-drug measure of reward, and apparently the brain opioid systems that mediate naltrexone-induced drinking suppression.     

Reversal of stress-induced analgesia by apomorphine,but not by amphetamine

Acute exposure to severe stressors induce profound analgesia as well as depleting catecholamine levels. The present study examined whether d-amphetamine and apomorphine, agents which increase catecholamine availability, would alter the analgesic effectiveness of cold-water swims (CWS) and 2-deoxy-D-glucose (2-DG) as measured by an operant liminal escape procedure. Two groups of 10 rats each were tested to determine alterations in liminal escape threshold functions following amphetamine at doses of 0.25, 0.5, 1,2 mg/kg and following apomorphine at doses of 0.025, 0.05, 0.1, 0.2 mg/kg. Half of the amphetamine and half of the apomorphine groups were tested across their respective dose ranges for the drug effects upon CWS analgesia. The remaining animals in each group received 2-DG (600 mg/kg IP) alone followed by 2-DG paired with each stimulant dose. No dose of amphetamine or apomorphine alone altered escape thresholds. While amphetamine produced slight potentiations of 2-DG analgesia at the two low doses, apomorphine at the 0.05 and 0.1 mg/kg doses returned CWS and 2-DG analgesia to within normal placebo values. These results provide indirect evidence for a role for brain norepinephrine and dopamine in stress-induced analgesia, and these data are discussed with respect to catecholamine involvement in pain-inhibitory processes.     

Differences in the pharmacokinetics of peroxisome proliferator-activated receptor agonists in genetically obese Zucker and sprague-dawley rats: implications of decreased glucuronidation in obese Zucker rats.

Genetically obese Zucker rats exhibit symptoms similar to those of obese patients with insulin-resistance or Type II diabetes; therefore, they have been used as a genetic model to study obesity, as well as a pharmacological model for the discovery of new drugs for the treatment of Type II diabetes and hyperlipidemia. In the present study, we compared the pharmacokinetics of two novel peroxisome proliferator-activated receptor (PPAR) agonists, MRL-I [(2R)-7-[3-[2-chloro-4-(4-fluorophenoxy)phenoxy]propoxy]-2-ethyl-3,4-dihydro-2H-benzopyran-2-carboxylic acid] and MRL-II [(2R)-7-[3-[2-chloro-4-(2,2,2-trifluoroethoxy)phenoxy]propoxy]-3,4-dihydro-2-methyl-2H-benzopyran-2-carboxylic acid], in obese Zucker and lean Sprague-Dawley rats following a single intravenous administration. The plasma clearance of both MRL-I and MRL-II was significantly lower in obese Zucker rats (4- and 2-fold, respectively) compared with Sprague-Dawley rats, but without any significant change in the volume of distribution, which resulted in a dramatic increase in the half-life (7- and 3-fold, respectively). The reversible in vitro plasma protein binding of [(14)C]MRL-I and [(14)C]MRL-II was comparable in the two strains, approximately 96% bound. The expression levels of uridine diphosphate-glucuronosyltransferases 1A1, 1A6, 2B1, and CYP2C11 and 3A1 mRNA in liver were lower (30-50%) in Zucker compared with Sprague-Dawley rats, as were the liver glutathione S-transferases (70%), quinone reductase (30%), organic anion-transporting protein 2 (80%), and multidrug resistance-associated protein 2 (Mrp2) (50%) mRNA levels. However, Mrp3 mRNA levels were similar in both strains. Consistent with these observations, the intrinsic clearance (CL(int)), calculated from the V(max)/K(m) of glucuronidation of [(14)C]MRL-I and [(14)C]MRL-II in liver microsomes, was approximately 2-fold lower in obese Zucker rats; the K(m) values were comparable in the two strains for both compounds. In conclusion, differences in the pharmacokinetics of two novel PPAR agonists, both cleared, predominantly, by conjugation, were evident in genetically obese Zucker rats compared with Sprague-Dawley rats. These differences were consistent with changes in the mRNA levels of hepatic drug-metabolizing enzymes and transporters. This information should be considered when comparing pharmacokinetic and efficacious doses in the obese Zucker rats, used as a pharmacological model, with those in Sprague-Dawley rats, which are used widely for drug metabolism and toxicology studies.     

Long-term physical exercise and atrial natriuretic peptide in obese Zucker rats

Endurance training increases natriuretic peptide synthesis in the hypertrophied myocardium of spontaneously hypertensive rats. We examined the effects of 22-week-long treadmill exercise on plasma and tissue atrial natriuretic peptide in Zucker rats, a model of genetic obesity and moderate hypertension without clear cardiac hypertrophy. The blood pressures of the animals were measured by the tail-cuff method, and plasma and tissue samples for the peptide determinations were taken at the end of the study. The training increased heart weight to body weight ratio, while atrial natriuretic peptide contents in the right and left atrium, ventricular tissue, and plasma did not change. The exercise prevented the elevation of blood pressure, which was observed in non-exercised obese Zucker rats, and also reduced blood pressure in the lean rats. In conclusion, these results suggest that in the absence of preceding myocardial hypertrophy, the long-term exercise-induced workload is not deleterious to the heart in experimental obesity, since no changes in plasma and tissue atrial natriuretic peptide were detected.     

Abolition of the expression but not the acquisition of latent inhibition by chronic amphetamine in rats

The animal amphetamine model of schizophrenia has been based primarily on stereotyped behavior. The present study sought to demonstrate an amphetamine-induced deficit in attentional processes. To this end, the effects of acute and chronic (14 days) 1.5 mg/kg dl-amphetamine administration on the ability of rats to ignore irrelevant stimuli were examined using the paradigm of latent inhibition (LI) in a conditioned emotional response (CER) procedure. The procedure consisted of three stages: pre-exposure, in which the to-be-conditoned stimulus, tone, was presented without being followed by reinforcement; acquisition, in which the pre-exposed tone was paired with shock; and test, in which LI was indexed by animals' suppression of licking during tone presentation. Experiment 1 showed that chronic but not acute treatment abolished LI. Experiment 2 showed that animals receiving chronic amphetamine pretreatment but pre-exposed and conditioned without the drug, exhibited normal LI. In Experiment 3, animals which received chronic amphetamine pretreatment and were pre-exposed under the drug but conditioned without it, also showed normal LI. The implications of these results for the animal amphetamine model of schizophrenia are discussed.     

Age-dependent changes in blood pressure and arterial reactivity in obese Zucker rats

The purpose of the present investigation was to determine whether there is an association between changes in arterial reactivity to vasoactive agents and the development of hypertension in obese Zucker rats. At 20 weeks of age, obese rats were mildly hypotensive compared to their lean littermate controls. Maximum contractile responses of endothelium-intact mesenteric arteries from these rats to noradrenaline, endothelin-1 and KCl were depressed, although there was no change in relaxation to acetylcholine. By 32 weeks of age, obese rats had developed hypertension compared to their lean littermate controls. Maximum contractile responses of mesenteric arteries from 32-week-old obese rats to noradrenaline and endothelin-1 were no longer significantly different than control, although contractile responses to KCl remained depressed. In addition, there was a small increase in sensitivity to endothelin-1, while endothelium-dependent relaxation to acetylcholine was impaired. In contrast, there were no changes in contractile responses of endothelium-intact aortas from either 20- or 32-week-old obese rats to noradrenaline, endothelin-1 or KCl, while endothelium-dependent relaxation of this artery to acetylcholine was slightly enhanced at both ages. Therefore, changes in the reactivity of the mesenteric artery but not the aorta from obese Zucker rats parallel changes in blood pressure in these animals.     

Early social isolation, but not maternal separation, affects behavioral sensitization to amphetamine in male and female adult rats.

Early life stressful manipulations, such as maternal separation (MS) or social isolation (SI), can influence the neurobiological development of rats and alter the response of adult animals to drugs of abuse. The present study examined the acute and sensitized behavioral responses (locomotor activity (LMA) and stereotypy) induced by amphetamine after MS or SI in male and female rats. In addition, the hypothesis that the combination of SI and MS could lead to additional effects on the behavioral response to amphetamine was tested. After the repetitive, intermittent administration of 1.5 mg/kg D-amphetamine over five consecutive days, the behavioral expression of sensitization to a challenge injection was assessed following a 2-day withdrawal period. In both sexes, MS and SI did not alter the acute locomotor activating effects of amphetamine as measured in the open-field environment after the first administration of the drug. Whereas SI altered the expression of sensitization to amphetamine in both sexes, MS did not affect it. Finally, in none of the behavioral variables measured did MS and SI interact to further modify the behavioral profile of the animals. The present results suggest that a postweaning manipulation of the environment (SI) is more effective than a preweaning manipulation (MS) in modifying the expression of sensitization to amphetamine.     

Triazolam attenuates amphetamine but not morphine conditioned place preferences

In a series of four experiments the benzodiazepine triazolam was tested for reinforcing effects and for effects on reinforcement induced by amphetamine and morphine. Reinforcement was assessed in a conditioned place preference paradigm. Triazolam did not produce reinforcing or aversive effects when administered in doses ranging from 0.0625 to 0.5 mg/kg. Triazolam did attenuate reinforcing effects produced by 0.75 and 1.25 mg/kg amphetamine. No effect of triazolam was observed on morphine-induced reinforcement. These results indicate that the administration of triazolam can affect the brain mechanisms that mediate the reinforcing effects of amphetamine but not morphine.     

Stress induced suppression of maintenance but not of acquisition of ethanol consumption in rats

Male Wistar rats were given a free-choice between water and increasing concentrations of ethanol from 3% to 11% (v/v) during the acquisition phase. Thereafter, animals were maintained on a choice between water and 11% ethanol for the balance of the experiment. For 5 days prior to and for periods during the acquisition and maintenance phases, the animals were expo exposed to electric footshock, restraint or no stress. The results showed no differences in ethanol drinking patterns among the groups during the acquisition phase. However, in the maintenance phase, both footshock and restraint suppressed the increase in ethanol intake seen in the no-stress control group.     

Disruption of selective attention by apomorphine,but not amphetamine,in the mongolian gerbil

To test the hypothesis that apomorphine, but not amphetamine, disrupts selective attention to a novel stimulus, gerbils were exposed to a novel object for one 60-s trial following an injection of 0, 1, 3, or 6 mg/kg d-amphetamine base, or 0, 0.1, 0.3, 1, 3, or 10 mg/kg apomorphine HClSC. They were tested the next day for habituation to the stimulus. As a control, half of each group of gerbils were injected but not exposed to the object on day 1. All non-exposed gerbils and all exposed gerbils that received amphetamine showed a decrement in investigation, indicative of habituation, on day 2. Furthermore, a gradient of responding during dishabituation was obtained from gerbils given d-amphetamine (1 mg/kg) which was dependent on the distance a novel object was moved, indicating a perception of location as occurs in normal gerbils. In contrast, those exposed gerbils that received 1 mg/kg or more of apomorphine did not show habituation on day 2. That the disruption of habituation by apomorphine was due to a failure of input rather than of retrieving the information was demonstrated in an experiment in which two groups of gerbils were habituated to a novel object prior to injection with apomorphine (1 mg/kg) or saline. Both groups continued to show habituation on subsequent trials and increased responding when the object was moved. Thus, the motor capabilities necessary for investigation were functional. When gerbils that received apomorphine were pretreated with the dopamine receptor blocker pimozide, habituation occurred on day 2, suggesting that the disruption of habituation was mediated by dopamine. On the other hand, the depressant effect of large doses of apomorphine on initial investigation was not blocked completely by pimozide.     

Effect of mevinolin on cholesterol metabolism in obese and lean Zucker rats

Mevinolin is a potent competitive inhibitor of HMG-CoA reductase, the enzyme catalyzing the major rate-limiting step in cholesterol synthesis. In this study the drug was administered as an intragastric dose at 2.5 mg/kg/day to 10 to 12-week-old lean and obese Zucker female rats over a 5-day period. Mevinolin showed no effect on plasma cholesterol levels in the lean rat; however, in the obese rat there was a significant decrease in plasma cholesterol (about a 40% decrease from initial levels). Although there was a difference in effect on plasma cholesterol levels in obese and lean rats, hepatocytes isolated from both fed lean and obese rats incubated with various concentrations of mevinolin exhibited similar levels of inhibition of cholesterol synthesis and showed no effects on the other metabolic processes studied. These results indicate that the drug was effective acutely on cholesterol synthesis in hepatocytes isolated from both lean and obese rats, but on a chronic treatment basis the hypocholesterolemic effect was observed only in the obese Zucker rat. This study supports the idea that the naturally occurring hypercholesterolemic obese Zucker rat may be a good model for testing potential new cholesterol lowering agents.     

Increase of anti-oxidation by exercise in the liver of obese Zucker rats

1. The effects of endurance training on the anti-oxidant status in diabetes were studied using obese Zucker rats. 2. We used a moderate exercise programme consisting of treadmill running at 20 m/min and 0% incline for 1 h/day, 7 days/week, for 8 weeks. At the end of the experimental period, changes in hepatic anti-oxidant enzymes in terms of protein content and mRNA levels were detected using western blotting analysis and northern blotting analysis, respectively. In addition, anti-oxidant enzyme activity was determined. 2. A significant reduction in mRNA levels and the protein content of hepatic Mn-superoxide dismutase (SOD) and glutathione peroxidase (GPx) were observed in non-exercise obese groups, but the mRNA and protein levels of these enzymes were markedly increased after exercise training. In addition, exercise training reversed the decreased enzyme activities of Mn-SOD and GPx in obese Zucker rats. 3. The diabetes-related lowering of the glutathione (GSH) concentration was elevated in exercised obese Zucker rats, indicating a marked effect of regular moderate exercise on the endogenous anti-oxidant system. 4. There were no marked changes in hepatic Cu/Zn-SOD in terms of mRNA levels, protein content and activity in sedentary obese Zucker rats compared with their lean littermates. Endurance training did not modify the gene expression and activity of hepatic Cu/Zn-SOD. 5. The results of the present study suggest that regular moderate exercise could improve the anti-oxidant defence function of Mn-SOD, GPx and GSH in obese Zucker rats.     

Sensitization to amphetamine,but not phencyclidine,disrupts prepulse inhibition and latent inhibition

Rationale Schizophrenia has been linked to dysregulation of dopamine and glutamate transmitter systems. Attempts to model aspects of schizophrenia in animals have made use of treatments that primarily affect dopaminergic (e.g., amphetamine, Amp) and glutamatergic (e.g., phencyclidine, PCP) function. In addition to exerting short-term acute effects, these agents also induce long-term effects, as seen, for example, in neurochemical and behavioural sensitization.Objectives The goal of this work was to compare Amp- and PCP-sensitized states on two measures of information processing that are impaired in schizophrenia, prepulse inhibition (PPI) of the acoustic startle reflex and latent inhibition (LI).Methods Rats received injections of Amp, PCP or saline 3 days per week for 3 weeks. The Amp dose increased from 1 to 3 mg/kg, at the rate of 1 mg/kg each week. The PCP dose was 3 mg/kg throughout. After various periods of withdrawal rats were tested for PPI and LI.Results Repeated intermittent treatment with Amp or PCP resulted in augmented locomotor responses to challenge with each drug, providing an operational index that sensitization had occurred. Rats sensitized to Amp showed disrupted PPI when tested drug free at 3, 21 and 60 days of withdrawal. Amp-sensitized rats also showed abolition of the LI effect. Rats sensitized to PCP did not show deficits in any of these behaviours when tested drug free.Conclusions Because disrupted PPI and LI have both been reported in schizophrenic patients, these results suggest that the Amp-sensitized state may represent a useful model for investigating the neural bases of information processing deficits in schizophrenia.