Limited Access Paradigm Research Articles

Mesolimbic exendin-4 attenuates reward salience evoked by neuropeptide Y and ghrelin

In the present study, we investigated the effects of the glucagon-like peptide-1 (GLP-1) agonist exendin-4 (Ex-4) on the stimulatory action of neuropeptide Y (NPY) and ghrelin. These effects were examined in relation to operant responding for palatable food or voluntary ethanol intake in a two-bottle limited access paradigm. Male Sprague Dawley rats, each with ventral tegmental area (VTA) unilateral guide cannulae, were used. Ex-4 was paired with either NPY, ghrelin, or combined NPY and ghrelin treatment. Our results indicated that while NPY and ghrelin reliably stimulated operant responding for sucrose pellets and increased ethanol intake, Ex-4 suppressed intake and, most importantly, significantly reduced the effects of NPY and ghrelin. Overall, this work provides compelling evidence that VTA GLP-1, NPY, and ghrelin systems interact within the brain to modulate reward salience.

Long-term alcohol drinking in High Drinking in the Dark mice is stable for many months and does not show alcohol deprivation effects.

We have modelled genetic risk for binge-like drinking by selectively breeding High Drinking in the Dark-1 and -2 (HDID-1 and HDID-2) mice for their propensity to reach intoxicating blood alcohol levels (BALs) after binge-like drinking in a single bottle, limited access paradigm. Interestingly, in standard two-bottle choice (2BC) tests for continuously available alcohol versus water, HDID mice show modest levels of preference. This indicates some degree of independence of the genetic contributions to risk for binge-like and sustained, continuous access drinking. We had few data where the drinking in the dark (DID) tests of binge-like drinking had been repeatedly performed, so we serially offered multiple DID tests to see whether binge-like drinking escalated. It did not. We also asked whether HDID mice would escalate their voluntary intake with prolonged exposure to alcohol 2BC. They did not. Lastly, we assessed whether an alcohol deprivation effect (ADE) developed. ADE is a temporary elevation in drinking typically observed after a period of abstinence from sustained access to alcohol choice. With repetition, these periods of ADE sometimes have led to more sustained elevations in drinking. We therefore asked whether repeated ADE episodes would elevate choice drinking in HDID mice. They did not. After nearly 500 days of alcohol access, the intake of HDID mice remained stable. We conclude that a genetically-enhanced high risk for binge-like drinking is not sufficient to yield alterations in long-term alcohol intake.

Gestational programming of ingestive behavior via effects on the mesolimbic dopaminergic reward pathway.

Obesity is a worldwide health issue that has reached epidemic proportions. While much of the research to this point has focused on discerning the role of hypothalamic feeding peptides in energy balance, this line of research has yielded little insight into how to resolve the obesity crisis. Therefore, it may be more productive to investigate other central neural systems that affect ingestive behavior, such as the mseolimbic dopaminergic reward system comprised of the ventral tegmental area (VTA) and nucleus accumbens (NAc). Recent research regarding the impact of gestational environment on energy balance indicates that gestational under‐ or over‐nutrition causes perturbations in hypothalamic neuropeptides responsible for regulating ingestive behavior and metabolism. This type of gestational programming may also affect the mesolimbic reward pathway, which serves to reinforce natural behaviors such as eating, drinking, and sexual activity. Because some research indicates that low birth weight (LBW) humans tend to exhibit both obesity and a propensity toward addiction, we hypothesized that maternal undernutrition during pregnancy may affect dopaminergic neuron function, which would impact the regulation of ingestive behavior. To test this hypothesis, we utilized a traditional rat model of intrauterine growth restriction by placing dams on a 50% food‐restricted diet during the second half of pregnancy. At PND1, litters were culled to 8 animals (4 male and 4 female) and cross‐fostered to control dams. At PND60 offspring were sacrificed by overdose of pentobarbital and perfused with saline followed by 4% paraformaldehyde. Afterward, brains were removed, post‐fixed in 4% paraformaldehyde for 48 hours, followed by a 30% sucrose sink. Brains were then sectioned at 35 microns and processed for immunohistochemical staining of tyrosine hydroxylase (TH), the rate‐limiting enzyme for dopamine production. Offspring of food restricted dams had significantly lower birth weights (5.6±0.08 g vs. 6.5±0.06 g, p<0.01) and significantly higher TH immunoreactivity in the VTA than controls (159±19 A.U. vs. 454±88 A.U., p<0.05). In a separate set of male and female PND90 offspring, we examined 24‐hour sucrose consumption as well as sucrose consumption in a limited access paradigm where animals received 10% sucrose for 90 minute intervals every 48 hours for one week. LBW offspring consumed significantly more sucrose during the limited access test (29±2 ml vs. 23±1 ml, p<0.05), but not when sucrose was provided with unlimited access over 24 hours. This suggests that LBW offspring exhibit enhanced ingestive behaviors when given limited access to rewarding substances. Consequently, the dopaminergic reward system appears to be a significant player in the underlying mechanism responsible for gestational programming related perturbations in ingestive behavior and energy balance. Further investigation into the contribution of reward pathway alterations that occur as a result of gestational programming could provide novel insight into potential ways to combat the obesity epidemic.Support or Funding InformationNSF CAREER 1350448

Effects of pair housing on voluntary alcohol intake in male and female Wistar rats.

A number of different voluntary alcohol intake paradigms are available for home cage drinking studies. Traditionally, these paradigms involve single housing in order for individual intake to be measured. This study aimed at investigating the effects of pair housing on voluntary alcohol intake. Male and female Wistar rats were housed in pairs or individually for studies of voluntary alcohol intake using the modified intermittent access paradigm with alcohol access during three consecutive days per week followed by four days of water only. Individual intake of 20% alcohol solution and water was measured during 12 sessions, i.e., 4 weeks. Pair-housed animals could interact freely with their cage mate for four consecutive days each week and were then separated by an inserted mesh divider for three consecutive days each week during alcohol intake sessions. Alcohol intake and preference were compared between pair-housed and individually housed rats. The results revealed higher alcohol intake in females than in males. Pair-housed males had a higher alcohol intake and preference during the first 3 weeks, but not during the fourth week, compared to individually housed males No effect of housing condition was observed in female rats. The alcohol intake was higher on the first day of access relative to the two consecutive days in pair-housed males and higher on the first two days relative to the third day in female rats. Social rank or female estrus cycle had no effect on alcohol intake or preference. Taken together, the use of a divider during alcohol intake sessions had no impact on alcohol intake in female rats and may not exert long-term influences in male rats. Future studies are needed in order to elucidate whether the use of a divider can constitute an experimental refinement as an alternative to individual housing in studies of voluntary alcohol intake using the limited access and/or intermittent access paradigms.

Corticostriatal Oscillations Predict High vs. Low Drinkers in a Rat Model of Limited Access Alcohol Consumption.

Individuals differ in their vulnerability to develop alcohol dependence, which is determined by innate and environmental factors. The corticostriatal circuit is heavily involved in the development of alcohol dependence and may contain neural information regarding vulnerability to drink excessively. In the current experiment, we hypothesized that we could characterize high and low alcohol-drinking rats (HD and LD, respectively) based on corticostriatal oscillations and that these subgroups would differentially respond to corticostriatal brain stimulation. Male Sprague–Dawley rats (n = 13) were trained to drink 10% alcohol in a limited access paradigm. In separate sessions, local field potentials (LFPs) were recorded from the nucleus accumbens shell (NAcSh) and medial prefrontal cortex (mPFC). Based on training alcohol consumption levels, we classified rats using a median split as HD or LD. Then, using machine-learning, we built predictive models to classify rats as HD or LD by corticostriatal LFPs and compared the model performance from real data to the performance of models built on data permutations. Additionally, we explored the impact of NAcSh or mPFC stimulation on alcohol consumption in HD vs. LD. Corticostriatal LFPs were able to predict HD vs. LD group classification with greater accuracy than expected by chance (>80% accuracy). Moreover, NAcSh stimulation significantly reduced alcohol consumption in HD, but not LD (p < 0.05), while mPFC stimulation did not alter drinking behavior in either HD or LD (p > 0.05). These data collectively show that the corticostriatal circuit is differentially involved in regulating alcohol intake in HD vs. LD rats, and suggests that corticostriatal activity may have the potential to predict a vulnerability to develop alcohol dependence in a clinical population.

Calcium chloride mimics the effects of acamprosate on cognitive deficits in chronic alcohol-exposed mice.

Acamprosate (calcium-bis N-acetylhomotaurinate) is the leading medication approved for the maintenance of abstinence, shown to reduce craving and relapse in animal models and human alcoholics. Acamprosate can improve executive functions that are impaired by chronic intermittent ethanol (CIE) exposure. Recent work has suggested that acamprosate's effects on relapse prevention are due to its calcium component, which raises the question whether its pro-cognitive effects are similarly mediated by calcium. This study examined the effects of acamprosate on alcohol-induced behavioral deficits and compared them with the effects of the sodium salt version of N-acetylhomotaurinate or calcium chloride, respectively. We exposed mice to alcohol via three cycles of CIE and measured changes in alcohol consumption in a limited-access paradigm. We then compared the effects of acamprosate and calcium chloride (applied subchronically for 3days during withdrawal) in a battery of cognitive tasks that have been shown to be affected by chronic alcohol exposure. CIE-treated animals showed deficits in attentional set-shifting and deficits in novel object recognition. Alcohol-treated animals showed no impairments in social novelty detection and interaction, or delayed spontaneous alternation. Both acamprosate and calcium chloride ameliorated alcohol-induced cognitive deficits to comparable extents. In contrast, the sodium salt version of N-acetylhomotaurinate did not reverse the cognitive deficits. These results add evidence to the notion that acamprosate produces its anti-relapse effects through its calcium moiety. Our results also suggest that improved regulation of drug intake by acamprosate after withdrawal might at least in part be related to improved cognitive function.

Oral self-administration of EtOH: sex-dependent modulation by running wheel access in C57BL/6J mice.

The effects of stress, including neuroendocrine and behavioral sequelae aimed at maintaining homeostasis, are associated with increased alcohol consumption. Because both stress and drinking are multifactorial, the mechanisms underlying the relationship are difficult to elucidate. We therefore employed an animal model investigating the influence of blocked access to a running wheel on drinking in C57BL/6J (B6) mice. In the first experiment, naïve, adult male and female subjects were individually housed for 2 weeks with 24-hour access to a running wheel and 12% ethanol (EtOH) in a 2-bottle, free choice paradigm. After determining baseline consumption and preference, experimental subjects had the running wheel placed in a locked position for 6 hours, and the EtOH bottle was removed during the first half of this period. Two subsequent experiments, again in adult, naïve B6 mice, examined the influence of locked running wheels on self-administration of 20% EtOH in a limited access paradigm, and blood EtOH concentrations (BECs) were determined on the final day of this protocol. In all 3 studies, using both between- and within-subject analyses, females showed transient yet reliable increases in alcohol drinking during blocked access to a rotating activity, while drinking in male mice was largely insensitive to this manipulation, although both sexes showed appreciable BECs (>130 mg/dl in females and 80 mg/dl in males) following a 2-hour EtOH access period. These data add to a burgeoning literature suggesting that the factors contributing to excessive alcohol use differ between males and females and that females may be especially sensitive to the influence of wheel manipulation. Elucidating the sex-dependent mechanisms mediating differences in alcohol sensitivity and response is critical to understanding the causes of alcoholism and in developing effective treatments and interventions.

P-28 * OPIOIDERGIC MODULATION OF ETHANOL INTAKE IN THE NUCLEUS ACCUMBENS OF ALCOHOL-PREFERRING AA RATS

The present study addressed the role of accumbal mu- and kappa-opioid receptors in ethanol intake using male AA rats trained to drink 10% (v/v) ethanol in a limited access paradigm. The mu-opioid receptor antagonist CTOP (0.3, 1, or 3 ug per side), the mu-opioid receptor agonist DAMGO (0.03, 0.1, or 0.3 ug), the kappa-opioid receptor agonist U50488H (0.3, 1, or 3 ug), and the kappa-opioid receptor antagonists JDTic (15 ug) and nor-binaltorphimine (3 ug) were injected bilaterally into the nucleus accumbens shell. In another experiment, rats received a single subcutaneous injection of JDTic (10 mg/kg) and the long-term effects of the treatment on ethanol intake were monitored. CTOP, DAMGO and U50488H failed to alter ethanol intake significantly, though CTOP (0.3 and 1 ug) tended to increase and DAMGO (0.1 ug) to decrease ethanol intake. There was a tendency for decreased ethanol intake 72 h after accumbal JDTic, while systemic JDTic decreased ethanol intake at 48 h and 96 h. These results support the notion that central mu- and kappa-opioid receptors participate in the control of voluntary ethanol intake. They also suggest that accumbal kappa-opioid receptors may have a more prominent role in ethanol intake than previously assumed.

Role of CA2+/calmodulin on ethanol neurobehavioral effects.

The cAMP-dependent protein kinase A (PKA) signaling transduction pathway has been shown to play an important role in the modulation of several ethanol-induced behaviors. Different studies have demonstrated intracellular calcium (Ca(2+))-dependent activation of the PKA cascade after ethanol administration. Thus, the cAMP cascade mediator Ca(2+)-dependent calmodulin (CaM) has been strongly implicated in the central effects of ethanol. In this study, we assessed the role of the CaM inhibitor W7 on ethanol-induced stimulation, ethanol intake, and ethanol-induced activation of PKA. Swiss mice were pretreated with W7 (0-10mg/kg) 30min before ethanol (0-3.75g/kg) administration. Immediately, animals were placed during 20min in an open-field chamber. Ethanol (10%, v/v) intake in 2h was assessed using a limited access paradigm. Experiments with caffeine (0-15mg/kg), cocaine (0-4mg/kg), and saccharine (0.1%, w/v) were designed to compare their results to those obtained with ethanol. Western blot was assayed 45min after ethanol administration. Results showed that pretreatment with W7, reduced selectively in a dose-dependent fashion ethanol-induced locomotor stimulation and ethanol intake. The ethanol-induced activation of PKA was also prevented by W7 administration. These results demonstrate that CaM inhibition resulted in a selective reduction of ethanol-stimulating effects and ethanol intake. The PKA activation induced by ethanol was blocked after the CaM blockade with W7. These results provide further evidence of the key role of cellular Ca(2+)-dependent pathways on the central effects of ethanol.

Effects of Galgunhaejutang on Alcohol Consumption in C57BL/6 Mice

Objectives : Galgunhaejutang extract (GHT) is a Traditional Korean medical herbal mixture that has been tradtionally used to treat alcohol consumption. This study was preformed to evaluate the effects of GHT on alcohol consumption in C57BL/6 mice. Methods : Sixty three C57BL/6 mice were alcohol dependence-induced by limited access paradigm. Water, GHT 0.688 g/kg (GHT-L), or GHT 3.45 g/kg (GHT-H) were administrated for 10 days. The amounts of alcohol consumption for 2 hours, water consumption for 22 hours, food intake for 24 hours and body weight were measured. Results : There weren't significant differences in 2 hours of alcohol consumption, 22 hours of water consumption, 24 hours of food intake and body weight for ten days between vehicle group and GHT-L or GHT-H group. Conclusions : Further studies employing multi-dose and long term administration of GHT (more than 10 days) might be of benefit.

Increased Extracellular Glutamate In the Nucleus Accumbens Promotes Excessive Ethanol Drinking in Ethanol Dependent Mice

Using a well-established model of ethanol dependence and relapse, this study examined adaptations in glutamatergic transmission in the nucleus accumbens (NAc) and their role in regulating voluntary ethanol drinking. Mice were first trained to drink ethanol in a free-choice, limited access (2 h/day) paradigm. One group (EtOH mice) received repeated weekly cycles of chronic intermittent ethanol (CIE) exposure with intervening weeks of test drinking sessions, whereas the remaining mice (CTL mice) were similarly treated but did not receive CIE treatment. Over repeated cycles of CIE exposure, EtOH mice exhibited significant escalation in drinking (up to ∼3.5 g/kg), whereas drinking remained relatively stable at baseline levels (2-2.5 g/kg) in CTL mice. Using in vivo microdialysis procedures, extracellular glutamate (GLUEX) levels in the NAc were increased approximately twofold in EtOH mice compared with CTL mice, and this difference was observed 7 days after final CIE exposure, indicating that this hyperglutamatergic state persisted beyond acute withdrawal. This finding prompted additional studies examining the effects of pharmacologically manipulating GLUEX in the NAc on ethanol drinking in the CIE model. The non-selective glutamate reuptake antagonist, threo-β-benzyloxyaspartate (TBOA), was bilaterally microinjected into the NAc and found to dose-dependently increase drinking in nondependent (CTL) mice to levels attained by dependent (EtOH) mice. TBOA also further increased drinking in EtOH mice. In contrast, reducing glutamatergic transmission in the NAc via bilateral injections of the metabotropic glutamate receptor-2/3 agonist LY379268 reduced drinking in dependent (EtOH) mice to nondependent (CTL) levels, whereas having a more modest effect in decreasing ethanol consumption in CTL mice. Taken together, these data support an important role of glutamatergic transmission in the NAc in regulating ethanol drinking. Additionally, these results indicate that ethanol dependence produces adaptations that favor elevated glutamate activity in the NAc which, in turn, promote excessive levels of ethanol consumption associated with dependence.

Reverse Pharmacogenetic Modulation of the Nucleus Accumbens Reduces Ethanol Consumption in a Limited Access Paradigm

Bilateral stereotactic lesioning of the nucleus accumbens (NAc) core reduces relapse rates in alcohol-dependent patients but may cause irreversible cognitive deficits. Deep brain stimulation has similar effects but requires costly implanted hardware and regular surgical maintenance. Therefore, there is considerable interest in refining these approaches to develop reversible, minimally invasive treatments for alcohol dependence. Toward this end, we evaluated the feasibility of a reverse pharmacogenetic approach in a preclinical mouse model. We first assessed the predictive validity of a limited access ethanol consumption paradigm by confirming that electrolytic lesions of the NAc core decreased ethanol consumption, recapitulating the effects of similar lesions in humans. We then used this paradigm to test the effect of modulating activity in the NAc using the Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) hM3Dq and hM4Di. We found that increasing activity with hM3Dq had no effect, but suppressing activity with hM4Di reduced alcohol consumption to a similar extent as lesioning without affecting consumption of water or sucrose. These results may represent early steps toward a novel neurosurgical treatment modality for alcohol dependence that is reversible and externally titratable, yet highly targetable and less invasive than current approaches such as lesioning or deep brain stimulation.

Disturbances in behavior and cortical enkephalin gene expression during the anticipation of ethanol in rats characterized as high drinkers

The process of ethanol anticipation is a particularly important phenomenon that can determine subsequent drug-taking behavior. Recent studies suggest that systems within the medial prefrontal cortex (mPFC), during anticipation, may contribute to the goal-directed seeking of ethanol. The current investigation examined the possibility that the opioid peptide enkephalin (ENK), known to mediate some of the reinforcing properties of ethanol, may function in the mPFC during the anticipation of ethanol access. Using a limited access (3 h/d) paradigm for 10 days with 20% ethanol, Sprague-Dawley rats were first identified either as low drinkers (LD, <1.0 g/kg/3 h) or as high drinkers (HD, >2.0 g/kg/3 h) that exhibited a long-term phenotype of high ethanol consumption and a significant ethanol deprivation effect. During the anticipation period immediately preceding daily ethanol access, the HD rats compared to LD or Control animals with ad libitum ethanol access exhibited increased anticipatory behaviors, including greater exploratory behavior in a novel open field as revealed by significantly more time spent in the rearing position (+53–65%, p < 0.05) and increased number of rears made (+33–44%, p < 0.05) and greater novelty-seeking behavior in a hole-board apparatus revealed by an increase in total (+50–52%, p < 0.05) and novel nose pokes (+45–48%, p < 0.05). In the HD rats, analysis of the mPFC using real-time quantitative PCR showed significantly greater mRNA levels of ENK (p < 0.05) and the mu-opioid receptor (MOR) (p < 0.05), but not delta-opioid receptor (DOR), and this increase in ENK expression was found, using in situ hybridization, to occur specifically in the prelimbic (PrL) subregion of the mPFC. When injected into the PrL during the anticipation period, a MOR agonist but not DOR agonist significantly increased consumption of 20% ethanol (p < 0.05). These findings support the role of ENK, acting through MOR within the PrL to promote the anticipation and excessive consumption of ethanol.

The interaction of chronic restraint stress and voluntary alcohol intake: Effects on spatial memory in male rats

Alcohol consumption and exposure to stressful life events activate similar neural pathways and thus result in several comparable physiological and behavioral effects. Alcoholics in treatment claim that life stressors are the leading cause of continued drinking or relapse. However, few studies have investigated the interactive effects of stress and alcohol on cognitive behavior. The effects of restraint stress, alcohol, and stress in combination with alcohol were examined on a spatial memory test, the object placement (OP) task. In addition, intake levels were measured to determine if stress altered general consumption of alcohol. Male Sprague-Dawley rats were assigned to one of four conditions: no alcohol/no stress control (CON), stress alone (STR), alcohol alone (ALC), and STR+alcohol (STR+ALC). Following each restraint stress bout, the STR+ALC and the ALC groups were given access to 8% alcohol for 1h using the two-bottle choice limited access paradigm. As predicted, the STR+ALC group significantly increased alcohol consumption, while the ALC group had consistent drinking over the 10-day treatment. On the OP task, STR and ALC groups performed at chance levels, whereas the CON and STR+ALC groups significantly discriminated between objects in the new and old locations. These data show that stress increases alcohol intake and the intake of alcohol is associated with reduction of the stress-induced impairment of spatial memory. The data have important implications for the development of alcohol abuse and its treatment.

NPY receptor subtype specification for behavioral adaptive strategies during limited food access

The neuropeptide Y (NPY) system in the brain regulates a wide variety of behavioral, metabolic and hormonal homeostatic processes required for energy balance control. During times of limited food availability, NPY promotes behavioral hyperactivity necessary to explore and prepare for novel food resources. As NPY can act via 5 different receptor subtypes, we investigated the path through which NPY affects different behavioral components relevant for adaptation to such conditions. We tested NPY Y1 and Y2 receptor knockout mice and their wild-type littermate controls in a daily scheduled limited food access paradigm with unlimited access to running wheel. Here we show that NPY Y1 receptor deficient mice lack the expression of appetitive behavior and that NPY Y2 receptors control the level of hyperactive behavior under these conditions. Thus, receptor specificity determines the differential expression of NPY-mediated behavioral adaptations to overcome a negative energy status.

Opioidergic Modulation of Ethanol Self‐Administration in the Ventral Pallidum

Striatopallidal medium spiny neurons have been viewed as a final common path for drug reward and the ventral pallidum as an essential convergent point for hedonic and motivational signaling in the brain. The medium spiny neurons are GABAergic, but they colocalize enkephalin. Purpose of this study was to investigate the role of the opioidergic mechanisms of the ventral pallidum in ethanol self-administration behavior. Effects of bilateral microinjections of μ-, δ-, and κ-opioid receptor agonists and antagonists into the ventral pallidum on voluntary ethanol consumption were monitored in alcohol-preferring Alko Alcohol (AA) rats using the 90-minute limited access paradigm. Stimulation of μ-opioid receptors with DAMGO (0.01 to 0.1 μg) or morphine (1 to 10 μg) in the ventral pallidum decreased ethanol intake dose-dependently. Conversely, blocking μ-receptors with CTOP (0.3 to 3 μg) increased ethanol intake significantly. Unlike CTOP, DAMGO also increased locomotor activity. Consumption of ethanol was not modified significantly by a broad-spectrum opioid receptor antagonist naltrexone, by δ-opioid receptor agonist DPDPE or antagonist naltrindole, or by a κ-opioid receptor agonist U50,488H or antagonist nor-BNI. The study provides evidence for μ- but not δ- or κ-opioid receptors in the ventral pallidum playing a role in the regulation of voluntary ethanol consumption. Furthermore, present findings give support to earlier work, suggesting an essential role of pallidal opioidergic transmission in drug reward.

The Effects of Gonadectomy on Age- and Sex-Typical Patterns of Ethanol Consumption in Sprague-Dawley Rats

Ethanol intake levels characteristic of adult males and females emerge postpubertally. The present set of experiments examined the consequences of prepubertal and adult gonadectomies to explore whether the presence of gonadal hormones at puberty exerts organizational influences and/or plays an activational role in age- and sex-typical patterns of ethanol consumption. Male and female Sprague-Dawley rats were gonadectomized (GX), received sham gonadectomy (SH), or were left nonmanipulated (NM) at 1 of 2 ages, either prepubertally on postnatal day (P) 23 (early) or postpubertally in adulthood on P70 (late). Early surgery animals were tested for ethanol consumption either during adolescence (P28 to 39) or in adulthood at the same age that late surgery animals were tested (P75 to 86). Voluntary ethanol consumption was indexed using a 2-hour limited-access paradigm, with access to 2 bottles: one containing water and the other a sweetened ethanol solution. Age of GX did not impact patterns of ethanol consumption. Removal of testicular hormones in males, regardless of age of removal, elevated consumption levels in adulthood to female-typical levels. Ovariectomy did not have notable effects on ethanol drinking in females. Ethanol intake and preference of early SH males were significantly greater than those of both late SH and NM males. Removal of the gonads prior to puberty did not influence ethanol drinking or preference during adolescence in either males or females. These results suggest that testicular hormones play an activational role in lowering ethanol intake and preference of adult male rats. Pubertal hormones, in contrast, were found to exert little influence on ethanol drinking or preference during adolescence, although the effect of surgical manipulation itself during development was found to exert a long-lasting facilitatory effect on ethanol consumption in adulthood.

Opposite effects on the ingestion of ethanol and sucrose solutions after injections of muscimol into the nucleus accumbens shell

Injection of the GABA A receptor agonist muscimol into the nucleus accumbens shell (AcbSh) elicits robust feeding in satiated rats, but has no effect on water intake. The current study was designed to examine whether intra-AcbSh muscimol injections influence the intake of ethanol solutions in rats trained to drink using a limited access paradigm. We confirmed that bilateral injections of muscimol (100 ng) into the AcbSh produce large increases in the intake of sucrose solutions and of the chow maintenance diet but found in two independent experiments that these injections potently reduce the intake of a 10% ethanol solution. Furthermore, intra-AcbSh muscimol significantly increased intake of an ethanol–sucrose mixture. These results demonstrate that activating GABA A receptors in the vicinity of the AcbSh can have opposite effects on the intake of different caloric substances and are consistent with the possibility that GABAergic circuits in the AcbSh may play a role in mediating voluntary ethanol intake.

The interactive effects of methylphenidate and ethanol on ethanol consumption and locomotor activity in mice

The concomitant use of alcohol (EtOH) and the psychotherapeutic agent dl-methylphenidate (MPH) has risen as a consequence of an increase in ADHD diagnoses within the drinking age population. It was recently found that the combination of MPH and EtOH increases the self-report of pleasurable feelings relative to MPH alone. This finding raises concerns regarding the combined abuse liability for these two widely used drugs. The present behavioral study reports on the development of an adult male C57BL/6J (B6) mouse model to further characterize this MPH–EtOH interaction. We examined the effects of MPH on EtOH consumption in a limited access paradigm and EtOH stimulation of locomotor activity. B6 mice consumed about 2 g/kg EtOH daily and MPH dose-dependently reduced drinking. The most effective dose of MPH was 1.25 mg/kg, which produced a 41% decrease in drinking and had no effect on locomotor activity. However, when the 1.25 mg/kg dose of MPH was combined with a stimulatory dose of ethanol (1.75 g/kg) by intraperitoneal injection, there was a significantly enhanced stimulation of locomotor activity. The drug combination increased activity compared to the vehicle or MPH injections by 45% and increased the activity relative to EtOH alone by an additional 25%. The results of the EtOH and MPH interactions observed with the mouse model appear to be behaviorally relevant and suggest several converging mechanisms that may underlie MPH–EtOH interactions.

Differential involvement of the opioid receptor antagonist naloxone in motivational and hedonic aspects of reward

In the present study dose-dependent effects of the opioid receptor antagonist naloxone were investigated on the rewarding effects of sweetened condensed milk (SCM) in four behavioral paradigms addressing hedonic, consummatory as well as motivational aspects of a reward: odour-conditioned pleasure attenuation of the acoustic startle response (PAS), conditioned place preference (CPP), voluntary consumption in a limited access paradigm, as well as break point determination in a progressive ratio (PR) task. A dose-dependent reduction in reward-related behavior was observed in all paradigms, with exception of the break point in the PR task, which was not affected by naloxone at all. CPP for SCM was only affected by the highest dose of naloxone. The present results indicate that naloxone is more effective in suppressing the hedonic than motivational aspects of reward, further supporting the involvement of the endogenous opioid system in the mediation of hedonic properties of food reward.