Noah R. Gubner

D4 receptor deficiency in mice has limited effects on impulsivity and novelty seeking

Alleles of the human dopamine D(4) receptor (D(4)R) gene (DRD4.7) have repeatedly been found to correlate with novelty seeking, substance abuse, pathological gambling, and attention-deficit hyperactivity disorder (ADHD). If these various psychopathologies are a result of attenuated D(4)R-mediated signaling, mice lacking D(4)Rs (D(4)KO) should be more impulsive than wild-type (WT) mice and exhibit more novelty seeking. However, in our study, D(4)KO and WT mice showed similar levels of impulsivity as measured by delay discounting performance and response inhibition on a Go/No-go test, suggesting that D(4)R-mediated signaling may not affect impulsivity. D(4)KO mice were more active than WT mice in the first 5 min of a novel open field test, suggesting greater novelty seeking. For both genotypes, more impulsive mice habituated less in the novel open field. These data suggest that the absence of D(4)Rs is not sufficient to cause psychopathologies associated with heightened impulsivity and novelty seeking.

Accentuating effects of nicotine on ethanol response in mice with high genetic predisposition to ethanol-induced locomotor stimulation

BACKGROUND Co-morbid use of nicotine-containing tobacco products and alcohol is prevalent in alcohol dependent individuals. Common genetic factors could influence initial sensitivity to the independent or interactive effects of these drugs and play a role in their co-abuse. METHODS Locomotor sensitivity to nicotine and ethanol, alone and in combination, was assessed in mice bred for high (FAST) and low (SLOW) sensitivity to the locomotor stimulant effects of ethanol and in an inbred strain of mouse (DBA/2J) that has been shown to have extreme sensitivity to ethanol-induced stimulation in comparison to other strains. RESULTS The effects of nicotine and ethanol, alone and in combination, were dependent on genotype. In FAST and DBA/2J mice that show high sensitivity to ethanol-induced stimulation, nicotine accentuated the locomotor stimulant response to ethanol. This effect was not found in SLOW mice that are not stimulated by ethanol alone. CONCLUSIONS These data indicate that genes underlying differential sensitivity to the stimulant effects of ethanol alone also influence sensitivity to nicotine in combination with ethanol. Sensitivity to the stimulant effects of nicotine alone does not appear to predict the response to the drug combination, as FAST mice are sensitive to nicotine-induced stimulation, whereas SLOW and DBA/2J mice are not. The combination of nicotine and ethanol may have genotype-dependent effects that could impact co-abuse liability.

Racial differences in the relationship between rate of nicotine metabolism and nicotine intake from cigarette smoking.

Rate of nicotine metabolism has been identified as an important factor influencing nicotine intake and can be estimated using the nicotine metabolite ratio (NMR), a validated biomarker of CYP2A6 enzyme activity. Individuals who metabolize nicotine faster (higher NMR) may alter their smoking behavior to titrate their nicotine intake in order to maintain similar levels of nicotine in the body compared to slower nicotine metabolizers. There are known racial differences in the rate of nicotine metabolism with African Americans on average having a slower rate of nicotine metabolism compared to Whites. The goal of this study was to determine if there are racial differences in the relationship between rate of nicotine metabolism and measures of nicotine intake assessed using multiple biomarkers of nicotine and tobacco smoke exposure. Using secondary analyses of the screening data collected in a recently completed clinical trial, treatment-seeking African American and White daily smokers (10 or more cigarettes per day) were grouped into NMR quartiles so that the races could be compared at the same NMR, even though the distribution of NMR within race differed. The results indicated that rate of nicotine metabolism was a more important factor influencing nicotine intake in White smokers. Specifically, Whites were more likely to titrate their nicotine intake based on the rate at which they metabolize nicotine. However, this relationship was not found in African Americans. Overall there was a greater step-down, linear type relationship between NMR groups and cotinine or cotinine/cigarette in African Americans, which is consistent with the idea that differences in blood cotinine levels between the African American NMR groups were primarily due to differences in CYP2A6 enzyme activity without titration of nicotine intake among faster nicotine metabolizers.

Use of multiple tobacco products in a national sample of persons enrolled in addiction treatment

OBJECTIVE To explore use of tobacco products in relationship to marketing exposure among persons in addiction treatment. METHOD A random sample of treatment programs was drawn from the National Institute on Drug Abuse (NIDA) Clinical Trials Network (CTN). Participants in each program completed surveys concerning use of tobacco products (N=1113). Exposure to tobacco marketing and counter-marketing, advertising receptivity, and perceived health risks of smoking were tested for their association with use of multiple tobacco products. RESULTS Prevalence of combustible cigarette use was 77.9%. Weekly or greater use of other products was: e-cigarettes (17.7%), little filtered cigars (8.6%), smokeless tobacco (5.2%), and standard cigars (4.6%) with 24.4% using multiple tobacco products. Compared to single product users, multiple product users smoked more cigarettes per day (OR=1.03, 95% CI 1.01-1.05, p<0.001), were more likely to have tried to quit (OR=1.41, 95% CI 1.02-1.96, p=0.041), reported greater daily exposure to advertising for products other than combustible cigarettes (OR=1.93, CI 1.35-2.75, p<0.001), and greater daily exposure to tobacco counter-marketing (OR=1.70, 95% CI: 1.09-2.63, p=0.019). CONCLUSION Heavier smokers and those trying to quit may be more likely to use e-cigarettes, little filtered cigars, or smokeless tobacco and have greater susceptibility to their advertising. This highlights the importance of regulating advertising related to smoking cessation as their effectiveness for this purpose has not been demonstrated.

Muscarinic type 2 receptors in the lateral dorsal tegmental area modulate cocaine and food seeking behavior in rats.

The cholinergic input from the lateral dorsal tegmental area (LDTg) modulates the dopamine cells of the ventral tegmental area (VTA) and plays an important role in cocaine taking. Specific pharmacological agents that block or stimulate muscarinic receptors in the LDTg change acetylcholine (ACh) levels in the VTA. Furthermore, manipulations of cholinergic input in the VTA can change cocaine taking. In the current study, the ACh output from the LDTg was attenuated by treatment with the selective muscarinic type 2 (M2) autoreceptor agonist oxotremorine.sesquifumarate (OxoSQ). We hypothesized that OxoSQ would reduce the motivation of rats to self-administer both natural and drug rewards. Animals were tested on progressive ratio (PR) schedules of reinforcement for food pellets and cocaine. On test days, animals on food and on cocaine schedules were bilaterally microinjected prior to the test. Rats received either LDTg OxoSQ infusions or LDTg artificial cerebrospinal fluid (aCSF) infusions in a within-subjects design. In addition, infusions were delivered into a dorsal brain area above the LDTg as an anatomical control region. OxoSQ microinjection in the LDTg, compared to aCSF, significantly reduced both the number of self-administered pellets and cocaine infusions during the initial half of the session; this reduction was dose-dependent. OxoSQ microinjections into the area just dorsal to the LDTg had no significant effect on self-administration of food pellets or cocaine. Animals were also tested in locomotor activity chambers for motor effects following the above microinjections. Locomotor activity was mildly increased by OxoSQ microinjection into the LDTg during the initial half of the session. Overall, these data suggest that LDTg cholinergic neurons play an important role in modifying the reinforcing value of natural and drug rewards. These effects cannot be attributed to significant alterations of locomotor behavior and are likely accomplished through LDTg muscarinic autoreceptors.

Effects of nicotine on ethanol-induced locomotor sensitization: A model of neuroadaptation

Co-morbid use of nicotine-containing tobacco products and alcohol (ethanol) is prevalent in young adults initiating use and in alcohol dependent adults, suggesting that these drugs in combination may increase risk to develop dependence on one or both drugs. Neuroadaptations caused by repeated drug exposure are related to the development of drug dependence and vulnerability to relapse. Locomotor sensitization has been used as a behavioral measure used to detect changes in neural drug sensitivity that are thought to contribute to drug dependence and relapse. Locomotor sensitization was measured in the current studies to examine potential differences in the effects of nicotine and ethanol given alone and in combination. Baseline activity levels of DBA/2J mice were assessed on 2 days, then mice were treated for 10 days with saline, nicotine (1 or 2mg/kg of nicotine tartrate), ethanol (1 or 2g/kg), or nicotine plus ethanol and locomotor activity was assessed every third day. On the following day, all mice were challenged with ethanol to measure the expression of sensitization. Mice treated with both nicotine and ethanol exhibited greater stimulation than predicted from the combined independent effects of these drugs, consistent with our previously published results. The combined effects of nicotine and ethanol on locomotor sensitization were dependent on the dose of ethanol and whether testing was performed after the drugs were given together, or after challenge with ethanol alone. These results suggest that nicotine and ethanol in combination can have neuroadaptive effects that differ from the independent effects of these drugs.

Effects of Varenicline on Ethanol‐Induced Conditioned Place Preference, Locomotor Stimulation, and Sensitization

BACKGROUND Varenicline, a partial nicotinic acetylcholine receptor (nAChR) agonist, is a promising new drug for the treatment of alcohol (ethanol [EtOH]) dependence. Varenicline has been approved by the Food and Drug Administration as a smoking cessation therapeutic and has also been found to reduce EtOH consumption in humans and animal models of alcohol use. These studies examined the hypotheses that varenicline attenuates the stimulant and sensitizing effects of EtOH and reduces the motivational effects of EtOH-associated cues. The goal was to determine whether these effects of varenicline contribute to its pharmacotherapeutic effects for alcohol dependence. In addition, effects of varenicline on acute stimulation and/or on the acquisition of sensitization would suggest a role for nAChR involvement in these effects of EtOH. METHODS Dose-dependent effects of varenicline on the expression of EtOH-induced conditioned place preference (CPP), locomotor activation, and behavioral sensitization were examined. These measures model motivational effects of EtOH-associated cues, euphoric or stimulatory effects of EtOH, and EtOH-induced neuroadaptation. All studies used DBA/2J mice, an inbred strain with high sensitivity to these EtOH-related effects. RESULTS Varenicline did not significantly attenuate the expression of EtOH-induced CPP. Varenicline reduced locomotor activity and had the most pronounced effect in the presence of EtOH, with the largest effect on acute EtOH-induced locomotor stimulation and a trend for varenicline to attenuate the expression of EtOH-induced sensitization. CONCLUSIONS Because varenicline did not attenuate the expression of EtOH-induced CPP, it may not be effective at reducing the motivational effects of EtOH-associated cues. This outcome suggests that reductions in the motivational effects of EtOH-associated cues may not be involved in how varenicline reduces EtOH consumption. However, varenicline did have effects on locomotor behavior and significantly attenuated acute EtOH-induced locomotor stimulation. In humans who drink while taking varenicline, it might similarly reduce stimulant responses and have an impact on continued drinking. General sedative effects in such individuals should be carefully considered.

Unique genetic factors influence sensitivity to the rewarding and aversive effects of methamphetamine versus cocaine.

Genetic factors significantly influence addiction-related phenotypes. This is supported by the successful bidirectional selective breeding of two replicate sets of mouse lines for amount of methamphetamine consumed. Some of the same genetic factors that influence methamphetamine consumption have been previously found also to influence sensitivity to the conditioned rewarding and aversive effects of methamphetamine. The goal of the current studies was to determine if some of the same genetic factors influence sensitivity to the conditioned rewarding and aversive effects of cocaine. Cocaine conditioned reward was examined in methamphetamine high drinking and low drinking line mice using a conditioned place preference procedure and cocaine conditioned aversion was measured using a conditioned taste aversion procedure. In addition, a general sensitivity measure, locomotor stimulant response to cocaine, was assessed in these lines; previous data indicated no difference between the selected lines in sensitivity to methamphetamine-induced stimulation. In contrast to robust differences for methamphetamine, the methamphetamine high and low drinking lines did not differ in sensitivity to either the rewarding or aversive effects of cocaine. They also exhibited comparable sensitivity to cocaine-induced locomotor stimulation. These data suggest that the genetic factors that influence sensitivity to the conditioned rewarding and aversive effects of methamphetamine in these lines of mice do not influence sensitivity to these effects of cocaine. Thus, different genetic factors may influence risk for methamphetamine versus cocaine use.

Associations between binge drinking frequency and tobacco use among young adults

Tobacco use is greater among young adults who binge drink; yet there is limited research on tobacco use characteristics among different types of binge drinkers based on frequency. We aimed to characterize this relationship among young adults (18-25years old) who used both substances in the past month (smoked ≥1 cigarette, and drank ≥1 alcoholic beverage) using an anonymous online survey. Participants (N=1405, 65.0% male) were grouped based on binge drinking frequency and compared for tobacco use characteristics and demographics using bivariate analyses and multinomial logistic regression. Binge drinking frequency groups were: non-binge drinkers who consumed alcohol (0days; 27.5%); occasional (1-3days; 37.9%); intermediate (4-8days; 21.9%); and frequent (9+days; 12.7%) binge drinkers. Comparing each binge drinking group to non-binge drinkers: Both occasional and frequent binge drinkers smoked more cigarettes per day (p=0.001; p=0.002); frequent binge drinkers reported greater temptations to smoke in positive affective/social situations (p=0.02); intermediate binge drinkers were less likely to have a tobacco abstinence goal (p=0.03) but more likely to have made a serious tobacco quit attempt; all of the binge groups were more likely to be social smokers (all p<0.01). Overall, we also found a high rate of smoking on binge drinking days. Individuals smoked cigarettes on 85.7%±32.9% of days they binge drank. Extent of binge drinking (not just prevalence) is an important factor influencing smoking characteristics in young adults.

Electronic-cigarette use by individuals in treatment for substance abuse: A survey of 24 treatment centers in the United States

Prevalence and reasons for using electronic cigarettes (e-cigarettes) was examined among patients enrolled in 24 substance abuse treatment centers in the United States (N=1113). Prevalence of e-cigarette use was assessed for the full sample. Bivariate analyses and multivariate logistic regression were used to identify characteristics associated with e-cigarette use among current cigarette smokers (the majority of e-cigarette users). Overall 55.5% of the sample reported lifetime use of e-cigarettes, and 30.5% reported using e-cigarettes in the past 30days (current users). The main reasons for using e-cigarettes were (a) at times/places when smoking was prohibited (53.5%), and (b) as a way to quit/reduce cigarette smoking (50.3%). Daily vs non-daily e-cigarette users were more likely to use e-cigarettes both as a way to reduce health risks, and as a way to quit/reduce cigarette smoking. A majority of e-cigarette users (87.1%) reported dual use of e-cigarettes and tobacco cigarettes during the past month. Among current cigarette smokers, those that also used e-cigarettes smoked more cigarettes per day, were more likely to have made a past year cigarette quit attempt, and to have tried nicotine replacement therapy compared to cigarette only smokers. There was a high rate of dual e-cigarette and cigarette use by persons enrolled in addiction treatment. E-cigarette users may be heavier cigarette smokers trying to quit or reduce their cigarette smoking. However, e-cigarettes were also used at times when individuals could not smoke cigarettes. Substance abuse treatment centers developing tobacco policies need to consider these potentially conflicting reasons for using e-cigarettes.