Cristine L. Czachowski

Effect of operant self-administration of 10% ethanol plus 10% sucrose on dopamine and ethanol concentrations in the nucleus accumbens

Although operant ethanol self‐administration can increase accumbal dopamine activity, the relationship between dopamine and ethanol levels during consumption remains unclear. We trained Long‐Evans rats to self‐administer escalating concentrations of ethanol (with 10% sucrose) over 7 days, during which two to four lever presses resulted in 20 min of access to the solution with no further response requirements. Accumbal microdialysis was performed in rats self‐administering 10% ethanol (plus 10% sucrose) or 10% sucrose alone. Most ethanol (1.6 ± 0.2 g/kg) and sucrose intake occurred during the first 10 min of access. Sucrose ingestion did not induce significant changes in dopamine concentrations. Dopamine levels increased within the first 5 min of ethanol availability followed by a return to baseline, whereas brain ethanol levels reached peak concentration more than 40 min later. We found significant correlations between intake and dopamine concentration during the initial 10 min of consumption. Furthermore, ethanol‐conditioned rats consuming 10% sucrose showed no effect of ethanol expectation on dopamine activity. The transient rise in dopamine during ethanol ingestion suggests that the dopamine response was not solely due to the pharmacological properties of ethanol. The dopamine response may be related to the stimulus properties of ethanol presentation, which were strongest during consumption.

Building Bridges: The Transdisciplinary Study of Craving From the Animal Laboratory to the Lamppost

This article represents the proceedings of a symposium at the 2003 Research Society on Alcoholism meeting in Ft. Lauderdale, Florida, organized and chaired by Peter M. Monti. The presentations and presenters were (1) Alcohol Seeking and Self-Administration in Rats: The Role of Serotonin Activity, by Cristine L. Czachowski; (2) Assessing Binge Drinking in Monkeys, by Kathleen A. Grant; (3) Craving and the Perception of Time, by Michael Sayette; (4) Ecological and Laboratory Assessment of Alcohol Urges and Drinking: Effects of Naltrexone, by Peter M. Monti; and (5) Discussion, by Damaris J. Rohsenow.

Effects of systemic opioid receptor ligands on ethanol- and sucrose seeking and drinking in alcohol-preferring (P) and Long Evans rats

The endogenous opioid system has been implicated in mediating the reinforcing effects of ethanol (EtOH). Naltrexone (NTX), an opioid antagonist with concentration-dependent selectivity for the mu receptor, naltrindole (NTI), a selective delta receptor antagonist, and U50,488H, a selective kappa receptor agonist were examined in both alcohol-preferring (P) and nonselected (Long Evans (LE)) rats to determine whether they differentially affected the seeking and consumption of EtOH and sucrose. Using the sipper-tube model, rats reinforced with either 2 % sucrose or 10 % EtOH were injected with vehicle and either NTI (2.5, 5.0, or 10.0 mg/kg), U50 (2.5, 5.0, or 10.0 mg/kg), low-dose NTX (0.1, 0.3, or 1.0 mg/kg), or high-dose NTX (1.0, 3.0, or 10.0 mg/kg). Subsequent intakes (consummatory) or lever responses (seeking) were assessed. Overall, NTI, U50, and NTX attenuated intake and responding for sucrose and EtOH, with EtOH-reinforced P rats being the most sensitive to the effects of NTI on intake and seeking. U50 treatment decreased intake and seeking in both P and LE rats but did not selectively reduce EtOH intake or seeking in either line. P rats were more sensitive than LE rats to lower doses of NTX, and these doses more selectively attenuated responding for EtOH than sucrose. Higher doses of NTX suppressed intake and responding across both lines and reinforcers. These results suggest that drugs selective for the opioid receptors may be good pharmacotherapeutic targets, particularly in those with an underlying genetic predisposition for greater EtOH preference/intake.

Manipulations of Serotonin Function in the Nucleus Accumbens Core Produce Differential Effects on Ethanol and Sucrose Seeking and Intake

OBJECTIVE Behaviorally relevant stimuli, including alcohol, are processed through the nucleus accumbens/ventral tegmental area (VTA)/prefrontal cortex circuit. It is hypothesized that serotonin affects ethanol-directed behaviors by interacting with this system via projections from the dorsal raphe to the nucleus accumbens and VTA. The current studies utilized two different operant paradigms, one focusing on reinforcer seeking and one focusing on reinforcer self-administration (both with an ethanol and a sucrose solution as the reinforcer) to elucidate serotonin-specific regulation of these behaviors. METHODS The present experiments assessed the effects of microinjections of a serotonin1B agonist (CGS12066B) and a serotonin1A agonist (8-OH-DPAT) in the nucleus accumbens core on ethanol- and sucrose-reinforced seeking and intake. In four separate experiments, male Long-Evans rats were trained to complete a single response requirement that resulted in access to 10% ethanol or 2% sucrose for a 20-min drinking period. RESULTS Before microinjections, ethanol-reinforced subjects were consuming an average of 0.5-0.95 g/kg ethanol and making 50-100 responses during intermittent nonreinforced sham (no drug) sessions (sucrose groups had similar baseline response levels). In summary, findings from the four experiments showed the following: (1) manipulations of serotonin function that had effects on ethanol-reinforced responding had either no effect or less pronounced effects on sucrose-reinforced responding; (2) administration of the serotonin1B agonist decreased seeking behaviors to a greater degree than drinking behaviors; and (3) administration of the serotonin1A agonist decreased ethanol intake but not seeking with no impact at all on sucrose-reinforced behaviors. CONCLUSIONS Manipulations of serotonin activity in the nucleus accumbens core had little effect on sucrose-reinforced behaviors and differential effects on ethanol seeking versus intake, suggesting that this area may play a complex but selective role in the stimulus processing of external and internal alcohol-associated cues.

Increased delay discounting tracks with a high ethanol-seeking phenotype and subsequent ethanol seeking but not consumption.

BACKGROUND Increased levels of delay discounting have been associated with alcoholism and problematic levels of drinking. Attempts to assess the directionality of this relationship by studying individuals with a family history of alcoholism as well as rodent lines selectively bred for high home cage alcohol preference have yielded discordant results. One possible reason for this discordance is that increased levels of delay discounting may only track with specific processes that lead to addiction vulnerability. This study investigated this possibility by assessing 3 strains of rats previously identified to exhibit heritable differences in ethanol (EtOH) seeking and consumption. METHODS In an adjusting amount delay discounting task, alcohol-preferring (P) rats who display high levels of both EtOH seeking and consumption were compared to high alcohol-drinking (HAD2) rats who only exhibit moderate EtOH seeking despite high levels of consumption, and Long Evans (LE) rats who display moderate seeking and consumption. EtOH-seeking and consumption phenotypes were subsequently confirmed in an operant self-administration task with a procedural separation between EtOH seeking and drinking. RESULTS P rats discounted delayed rewards to a greater extent than both HAD2s and LE who did not show differences in discounting. Moreover, the EtOH-seeking and drinking phenotypes were replicated with P rats displaying greater EtOH seeking compared to both the HAD2s and LE, and both the HAD2s and P rats consuming more EtOH than LEs. CONCLUSIONS Only the high-seeking strain, the P rats, exhibited increased levels of delay discounting. This suggests that this measure of behavioral under-control is specifically associated with alcohol-related appetitive, but not consummatory, processes as the moderate seeking/high drinking line did not show increased levels of impulsivity. This finding supports the hypothesis that delay discounting is specifically associated with only certain processes which are sufficient but not necessary to confer addiction vulnerability and therefore also supports increased levels of delay discounting as a predisposing risk factor for alcoholism.

Alterations in ethanol seeking and self-administration following yohimbine in selectively bred alcohol-preferring (P) and high alcohol drinking (HAD-2) rats

Evidence suggests that stress increases alcohol drinking and promotes relapse in humans. Animal models that assess related behaviors include the sipper tube ethanol self-administration and the stress-induced reinstatement paradigms. While selectively bred for the same high-ethanol-drinking behavior, alcohol-preferring P rats appear to show greater sensitivity to ethanol reinforcement than high-alcohol-drinking HAD rats. The present experiment tested the effects of the pharmacological stressor, yohimbine, on the motivation to seek and consume ethanol implementing a combined sipper tube/reinstatement model using male P and HAD-2 rats. Following training to self-administer ethanol using the sipper tube procedure, rats were tested for the effects of yohimbine (0.625-2.5 mg/kg) on ethanol drinking. Subsequently, rats were tested for the effects of 1.25 mg/kg yohimbine on reinstatement of ethanol seeking. Yohimbine (0.625 and 1.25 mg/kg) increased ethanol self-administration, and the latter dose also decreased latency to complete the response requirement. Yohimbine elicited reinstatement of ethanol seeking in both lines. HAD-2 rats drank more ethanol, but showed similar responding on the ethanol-associated lever compared to P rats. These findings extend both the reinstatement and sipper tube models and justify further exploration of this unique combined paradigm. Despite prior evidence suggesting that P rats are more motivated to seek and consume ethanol, differences in these behaviors between P and HAD-2 rats were not systematic in the present experiment. Further investigation may elucidate whether either selected line may be more sensitive than other selectively bred or outbred rats to stress-related changes in ethanols reinforcing effects.

Tolcapone suppresses ethanol intake in alcohol preferring rats performing a novel cued access protocol

BACKGROUND Dopamine (DA) has been shown to play a central role in regulating motivated behavior and encoding reward. Chronic drug abuse elicits a state of hypodopaminergia in the mesocorticolimbic (MCL) system in both humans and preclinical rodent models of addiction, including those modeling alcohol use disorders (AUD). METHODS Working under the hypothesis that reductions in the bioavailability of DA play an integral role in the expression of the excessive drinking phenotype, the catechol-O-methyltransferase (COMT) inhibitor tolcapone was used as a means to amplify cortical DA concentration and drinking behaviors were then assessed. Sucrose and ethanol (EtOH) consumption were measured in P and Wistar rats in both a free choice drinking protocol and a novel cued access protocol. RESULTS Tolcapone attenuated the consumption of EtOH, and to a lesser extent sucrose, in P rats in the cued access protocol, while no effect was observed in the free choice drinking protocol. Tolcapone also decreased EtOH consumption in high drinking Wistar rats. A follow-up experiment using the indirect DA agonist d-amphetamine showed no change in EtOH consumption. CONCLUSIONS Collectively, these data suggest that COMT inhibitors may be capable of alleviating the extremely motivating or salient nature of stimuli associated with alcohol. The hypothesis is put forth that the relative specificity of tolcapone for cortical DA systems may mediate the suppression of the high seeking/drinking phenotype.

Measured Alcohol Content in College Party Mixed Drinks

The main objective of this study was to measure the alcohol content in college party drinks. Samples of mixed drinks were collected from on-campus parties (N = 23) over a 12-week period at a university in the Northeast. Samples were analyzed by using a method that measures oxygen utilization during ethanol oxidation. Standard drink equivalents were calculated and blood alcohol concentrations (BACs) for men and women were estimated. The percent alcohol in sampled drinks ranged from 3.7% to 22.8%. Characteristics of the sampled parties were not related to drink concentration. A party drink at the median concentration and drink size contained 0.97 standard drinks. Estimated BACs varied widely depending on drink alcohol concentration, but in most cases a heavy drinking episode for both men and women resulted in an estimated BAC at or above .08. Mixed drinks at the sampled parties on average approximated one standard drink, but the variability in mixed drink strength compromises a drinkers ability to keep track of the number of drinks consumed. (PsycINFO Database Record (c) 2009 APA, all rights reserved).

Effects of group II metabotropic glutamate receptor modulation on ethanol- and sucrose-seeking and consumption in the rat

Previous studies suggest that group II metabotropic glutamate receptors (mGluR2/3) are involved in regulating ethanol-seeking and consumption. The mGluR2/3 agonist LY379268 (LY37) and selective mGluR2 positive allosteric modulator biphenyl‑indanone A (BINA) were used to investigate the relative contribution of mGlu2 and mGlu3 receptors on ethanol- and sucrose-seeking and consumption. A microinjection study was then performed to examine the role of nucleus accumbens (NAc) core mGluR2/3 on ethanol-seeking. For the systemic experiments, separate groups of male Wistar rats [LY37 (0-2.0 mg/kg); BINA (0-20 mg/kg)] were trained to complete a response requirement (RR) resulting in access to 10% ethanol or 2% sucrose (in separate groups) for a 20‑min drinking period. Animals then underwent consummatory testing (weekly drug injections with RR1) followed by appetitive testing (weekly drug injections followed by extinction session). A separate group of male Wistar rats was surgically implanted with bilateral guide cannulae directed toward the NAc core and had weekly microinjections followed by an extinction session. Systemic administration of the mGluR2/3 agonist LY37 significantly reduced ethanol- and sucrose-seeking. The same treatment also reduced sucrose consumption and body weight (24‑h post injection). Systemic administration of the selective mGluR2 PAM BINA, however, had no effect on either seeking or consumption of ethanol or sucrose. Intra-accumbens core LY37 significantly reduced ethanol-seeking. These findings suggest that systemic mGluR2/3 agonism, but not allosteric modulation of mGluR2, reduces reinforcer-seeking. In particular, NAc core group II mGluR may be involved in regulating ethanol-seeking.