Thesis (Ph. D.)--University of Missouri--Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves : 107-109). Also available on the Internet.

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No. of bitstreams: 1 DissPP.pdf: 2336243 bytes, checksum: 6713ce2a544f0c9590312b61ae159504 (MD5) Previous issue date: 2017-05-12 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Context-induced reinstatement of drug seeking is an animal model for assessing the neural mechanisms underlying context-induced drug relapse, a major factor in human drug addiction. Neural activity in prelimbic cortex has been implicated in context-induced reinstatement drug seeking. Here, we report a new and feasible training procedure for context-induced reinstatement of alcohol seeking to explore the role of prelimbic cortex in context-induced relapse to alcohol seeking. Long-Evans rats were first given home-cage access to 10% ethanol. Using a saccharin fading technique, rats were first trained to self-administer 10% ethanol in one context. Next, lever pressing in the presence of the discrete cue was extinguished in a different context. Subsequently, context-induced reinstatement of drug seeking was assessed by re-exposing rats to the drug-associated or extinction context under extinction conditions. Re-exposure to the alcohol-associated context reinstated alcohol seeking and increased expression of the neural activity marker Fos in the prelimbic prefrontal cortex. The percentage of neural activation in this brain region was 3.4 % in extinction context and 7.7 % in drug associated context. Reversible inactivation of neural activity in prelimbic prefrontal cortex using GABA agonists muscimol and baclofen attenuated context-induced reinstatement. Based on previous findings of the literature that Fos-expressing neurons play a critical role in conditioned drug effects, we assessed whether context-induced reinstatement was associated with molecular alterations selectively induced within context-activated Fos-expressing neurons. We used fluorescence-activated cell sorting to isolate reinstatement-activated Fos-positive neurons from Fos-negative neurons in prelimbic cortex and used quantitative PCR to assess gene expression within these two populations of neurons. Context-induced reinstatement was associated with increased expression of the GABAα5 gene and decrease in the AMPA receptor subunit genes GluR1 and GluR2 in only Fos-positive neurons. Our results demonstrate an important role of prelimbic cortex in context-induced reinstatement of alcohol seeking and that this reinstatement is associated with unique gene alterations in Fos-expressing neurons. / Uma das principais dificuldades relacionada ao tratamento da dependência de etanol é o alto índice de reincidência ao uso dessa substância. Neste sentido, a exposição ao contexto associado ao uso de etanol é a principal causa de recaída. Evidencias mostram que a recaída envolve comportamentos de aprendizado associativo. Essas associações são armazenadas por pequenos grupamentos neurais conhecidos como “neuronal ensembles”. A recaída induzida pelo contexto pode ser estudada através de um modelo animal conhecido como “ABA renewal”. Neste procedimento, ratos são treinados a autoadministrarem etanol em um determinado contexto. Após o aprendizado, os roedores são submetidos ao procedimento de extinção desse comportamento, em um ambiente com diferenças táteis, visuais e sonoras do contexto do treino. A reinstalação é avaliada através da exposição do animal ao contexto inicial. O objetivo do presente estudo foi: a) comparar o desempenho de ratos Wistar e Long-Evans no protocolo de reinstalação da autoadministração de etanol induzida pelo ambiente; b) investigar a participação do córtex pré-frontal medial nesse comportamento; c) investigar a presença de alterações na expressão gênica em “neuronal ensembles” do córtex pré-límbico, relacionadas a reinstalação da autoadministração de etanol. Observamos quea exposição ao contexto previamente associado a autoadministração de etanol foi capaz de reinstalar esse comportamento em ratos Long-Evans e Wistar. Sendo que a linhagem de ratos Long-Evans apresentou melhor desempenho no teste de reinstalação da autoadministração de etanol induzida pelo contexto quando comparado aos ratos Wistar. Demonstramos que a reinstalação da autoadministração de etanol induzida pelo contexto promoveu aumento da ativação neuronal do córtex pré-límbico. Notamos ainda, que uma pequena porcentagem de neurônios foi ativada no córtex pré-límbico durante a reinstalação da autoadministração de etanol induzida pelo contexto (~7,7%). Evidenciamos que a inibição farmacológica do córtex pré-límbico atenuou a reinstalação da autoadministração de etanol induzida pelo contexto. E por fim, encontramos que a reinstalação da autoadministração de etanol foi associada a redução na expressão de GluR1 e GluR2 (subunidades do receptor glutamatérgico AMPA) e ao aumento na expressão de GABAα5 (subunidade do receptor GABAérgico GABA A) em neurônios seletivamente ativados no córtex pré-límbico durante o teste comportamental. Concluímos que a reinstalação da autoadministração de etanol induzida pelo ambiente é mediada por alterações na expressão gênica em “neuronal ensembles” ativados pela associação entre o efeito reforçador do etanol e o contexto em que essa substância de abuso foi administrada.

Le système endocannabinoïde (SEC) est composé : de deux neurotransmetteurs principaux qui sont l’anandamide (AEA) et le 2-arachidonoylglycerol (2-AG), de deux enzymes de catabolisme associées, respectivement la Fatty Acid Amide Hydrolase (FAAH) et la Monoacylglycerol Lipase (MAGL); et de deux récepteurs principaux qui sont les récepteurs cannabinoïdes de type 1 (CB1) et de type 2 (CB2). Le SEC exerce un rôle critique dans le contrôle des propriétés récompensantes des substances addictives, dont la nicotine. Cependant, le SEC possède un mode de fonctionnement biphasique et complexe. Par exemple, alors que les propriétés renforçantes et incitatrices de la nicotine sont diminuées par le blocage aigu des récepteurs CB1, le comportement de recherche de nicotine peut également être bloqué par l’augmentation aigüe du tonus endocannabinoïde (eCB). Par ailleurs, les essais cliniques suggèrent que le traitement chronique avec l’antagoniste des récepteurs CB1 produit des effets secondaires liés à l’état émotionnel des fumeurs abstinents. Ces résultats indiquent clairement, que l’utilisation optimale de la pharmacologie cannabinoïde pour le traitement chronique du sevrage tabagique reste compliquée et encore mal maitrisée. Le but de ce travail de thèse était d’abord de préciser le rôle des récepteurs CB1 dans le contrôle aigu des propriétés addictives de la nicotine, puis de déterminer le rôle de l’AEA dans le contrôle à long terme des propriétés incitatives de la nicotine. L’hypothèse générale était que la prise volontaire de nicotine, puis le traitement chronique avec un inhibiteur de la FAAH, produiraient une augmentation de l’AEA persistante chez le rat abstinent. Celle-ci aurait 2 conséquences : d’une part la réduction du comportement de recherche de nicotine, et d’autre part le développement d’un état anxieux généralisé indépendant de la transmission via les récepteurs CB1. Pour tester cette hypothèse, des rats ont été exposés à l’auto-administration intraveineuse de nicotine pendant 8 semaines puis mis en abstinence pendant 8 semaines. Pendant cette seconde période, les animaux ont été injectés quotidiennement avec un inhibiteur de la FAAH et nous avons caractérisé le comportement de recherche de nicotine et l’état émotionnel de ces rats. Les résultats ont d’abord montré une grande variabilité inter-individuelle dans la prise volontaire de nicotine, nous avons pu identifier des rats à faible consommation (« low consumers ») et des rats à forte consommation (« high consumers »). Il semble que l’inhibition chronique de la FAAH bloque significativement la rechute induite par la drogue et par les stimuli environnementaux chez les « low consumers » abstinents, et reste sans conséquence sur l’état émotionnel des rats. En revanche chez les « high consumers », l’inhibition de la FAAH bloque uniquement la rechute induite par la nicotine et ces animaux restent sensibles aux effets précipitants des stimuli environnementaux. Par ailleurs, ils développent un état anxieux modéré qui n’est pas bloqué par le traitement avec un antagoniste des récepteurs CB1. Ces données suggèrent donc que chez les « high consumers » abstinents, il existe un excès d’AEA qui cible des circuits différents pour moduler les propriétés incitatrices de la nicotine et l’anxiété. En conclusion, cette étude montre que l’inhibiteur de FAAH pourrait être un outil thérapeutique adapté au traitement de l’addiction à la nicotine si l’on prend en compte la variabilité inter-individuelle rencontrée dans la consommation abusive de nicotine. / The endogenous cannabinoid system, also called the endocannabinoid system (ECS), comprises two principal neurotransmitters: anandamide (AEA) and 2-arachidonoylglycerol (2-AG), whose specific degradation enzymes are fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), respectively. Both AEA and 2-AG bind to cannabinoid type 1 (CB1) and cannabinoid type 2 (CB2) receptors. The ECS is a key element for the expression of both natural and pharmacological reward processes, including nicotine reward. However, the biphasic and complex nature of the ECS renders it a difficult system to study. For example, although acute blockade of CB1 receptors reduces the reinforcing and incentive properties of nicotine, it has also been demonstrated that an acute increase of AEA (CB1 agonist) blocks nicotine-seeking behavior. The aim of the present work is twofold: first, to better characterize the neural substrates through which CB1 receptors regulate the voluntary intake of nicotine; and second, to determine the effect of a chronic increase of AEA tone in nicotine-abstinent rats on i) their persistent motivation for nicotine and ii) their affective phenotype. The hypothesis tested here is that chronic treatment with a FAAH inhibitor after chronic exposure to nicotine will induce a large and persistent increase of AEA during a period of nicotine abstinence. Subsequently, high levels of AEA would reduce the incentive properties of nicotine and nicotine-related stimuli, thus reducing nicotine seeking, and it would also promote high levels of anxiety which might be CB1-independent. In order to test this hypothesis, rats were exposed to nicotine (60 µg/kg/0.1 ml) intravenous self-administration (IVSA) for 8 weeks, after which they remained nicotine-free until the end of the experiment. During the period of abstinence, animals were injected daily with a FAAH inhibitor for 8 weeks (URB597, 0.3 mg/kg i.p.), and both their motivation for nicotine and their affective profile were assessed. Animals were classified into “low consumers” and “high consumers” since a high degree of inter-individual variability was observed in the total amount of nicotine taken over the 8-week IVSA period. We show that in the “low consumers”, chronic FAAH inhibition dramatically reduces nicotine seeking behavior and cue-related incentive salience, and does not induce anxiety-related side-effects during abstinence. In the “high consumers”, however, chronic FAAH inhibition reduces nicotine seeking but not cue-related incentive salience, and causes an increase in anxiety-like behaviors that are not blocked by a CB1 receptor antagonist. These data suggest that in the population of “high consumers”, the effects of increased AEA tone on motivation and anxiety might be mediated by different neuronal circuits. In conclusion, this study demonstrates the potential of chronic FAAH inhibition as an important therapeutic tool for the treatment of nicotine addiction in subjects with a moderate nicotine intake.

L’addiction aux drogues d’abus est un désordre psychiatrique caractérisé par une consommation compulsive de toxiques, au détriment d’activités alternatives plus bénéfiques à long terme, et malgré les conséquences négatives associées. Cette psychopathologie est ainsi de plus en plus conceptualisée comme le résultat d’une prise de décision pathologique, et l’un des plus grands enjeux de la recherche en addiction est de comprendre les bases neurobiologiques et physiopathologiques de ce dysfonctionnement. Devant les contraintes éthiques, techniques et pratiques inhérentes aux études neurobiologiques chez l’homme, il est important d’utiliser en parallèle des modèles animaux d’addiction. Malheureusement, la validité de la plupart de ces modèles est incertaine. En effet, la prise de drogue est bien trop souvent étudiée dans une situation où la drogue est la seule récompense disponible. Ainsi, dans ces conditions, il est difficile de déterminer si l’animal s’auto-administre une drogue par compulsion, ou simplement parce qu’il n’a pas d’autre activité alternative valable. Une série d’expériences menées dans notre laboratoire depuis plusieurs années a montré que lorsqu’un choix est donné entre une injection intraveineuse de cocaïne ou d’héroïne et un accès à une boisson sucrée, la vaste majorité des rats préfèrent l’eau sucrée. Une minorité de rats persévère cependant dans le choix de la drogue au détriment de l’accès à la boisson sucrée, et constituerait ainsi une population d’individus vulnérables à l’addiction. Au cours de ma thèse, mon principal objectif a été d’étudier les déterminants du choix entre prise de drogue (cocaïne et héroïne) et récompense alternative. Mes travaux de recherche ont permis de déterminer les mécanismes de choix utilisés par le rat, en testant la validité prédictive relative de plusieurs modèles de prise de décision. J’ai également contribué à la mise en évidence d’une interaction forte entre le contexte de choix, les effets directs de la drogue et les capacités cognitives spécifiques au rat. Cette interaction exerce une influence considérable sur le choix de la drogue chez le rat. Cette découverte pourrait conduire à une réinterprétation des expériences de choix menées chez le rat et soulève aussi des interrogations importantes sur la validité relative des procédures de choix pour modéliser l’addiction aux drogues. / Drug addiction is defined as a psychiatric disorder involving compulsive drug use, despite negative consequences, and is increasingly conceptualized as resulting from poor decision making with a preference bias towards the drug at the expense of other socially-valued behaviors. The most important challenge in current addiction research is to understand the physiopathology of this disorder. Animal models are important tools in addiction research, since they are less ethically and technically limited than human studies. However, preclinical research on drug addiction is typically performed in laboratory rats that are given ready access to drugs for intravenous self-administration but without other options. The lack of choice during drug access limits its validity for understanding the physiopathology of addiction. A series of studies from our laboratory has previously shown that when offered a mutually exclusive choice between pressing a lever to get sweet water or an alternative lever to receive an intravenous dose of cocaine or heroin, most rats prefer sweet water. Only a minority of rats persists in drug taking despite the availability of an alternative reward, and thus, appears to be more vulnerable to drug addiction. During my thesis, my main objective was to determine the psychological and behavioral determinants of choice between drugs of abuse (cocaine and heroin) and sweet water. This research allowed us to determine the decision-making processes underlying this choice, by testing the predictive validity of different decision-making models. Additionally, we evidenced a strong interaction between the choice setting, the drug’ direct effects and rats’ specific cognitive abilities that reliably influences drug choices in rats. This finding should lead to a novel interpretation of drug choice studies in rats and also raises important issues regarding the relative validity of choice procedures for modelling drug addiction.

Background: Studies with experimental administration of alcohol offer inconsistent approaches and interpretations in which ways an acute alcohol exposure affects the HPA-system and the cortisol secretion. So far published alcohol experiments differ in alcohol application, the possibility of alcohol self-administration at the subjects own discretion as well as the age of the participants. Question: Is cortisol secretion modified by gender during alcohol infusion? Do men and women show different cortisol levels under alcohol exposure when compared to the baseline? Is there a dose-response relationship between cortisol secretion and acute alcohol exposure? Have family history, smoking habits and alcohol induced side effects like nausea an influence on the cortisol secretion under alcohol exposure? Materials and methods: 48 18 year old subjects participated in two identical sessions in which they were able to regulate their maximum blood alcohol concentration up to a safety limit of 1.2 ‰ (i.e., 0.12%). The experiment was conducted by using a special software for self-infusion of ethanol (CASE) which guided the participants through a two and a half hours long experiment. CASE is founded on a validated physiologically-based pharmacokinetic model and involves calculating the infusion volume needed to increase the blood alcohol concentration in a linear manner. The BAC increases after each alcohol request by 0,075 ‰ (i.e., 0.0075%) within in two and a half minutes. If the subject infuses no alcohol the blood alcohol concentration will decrease by 0.01 ‰ (i.e., 0.001%) per minute. Through the precise calculation of the infusion rate, individual differences can be eliminated. Cortisol levels were measured at five time points: an initial baseline measurement and four measurements at fixed time points during the alcohol self-administration of subjects with two completed alcohol experiments. As an appropriate measure to examine the effect of alcohol self-administration on cortisol secretion, the maximum blood alcohol concentration was determined. In addition the day of experiment, gender and family history were observed as well as exploratory maximum nausea and smoking habits. Results: In conclusion, the results confirmed that women have higher cortisol levels than men at baseline and under alcohol influence. Blood alcohol concentration as examined influencing variable was shown to have different effects on the HPA system on day one and two. On the first day of experimentation there was no effect of blood alcohol concentration on the HPA system. On the second day a dose-response relationship could be identified between cortisol secretion and acute alcohol exposure. Individually higher blood alcohol concentrations attenuated cortisol stronger in comparison to subjects with lower blood alcohol concentrations. Family history, smoking habits and unpleasant side effects (nausea) did not affect the cortisol secretion under alcohol exposure in this series of experiments. Conclusions: Current data suggests that alcohol experiments affect the cortisol secretion in young social drinkers. These findings could be detected for the first time. Up to this point there has not been an experimental study that investigated and evaluated the dose-effect relationship between cortisol secretion and alcohol in a study design which uses intravenous alcohol self-administration. It can be theorized that the first day of experimentation is suitable as a settling-in phase due to unspecific confounding factors, whereas the second day can be considered, in an identical setting, apt for hypothesis testing. The increased cortisol level in women when compared to men is consistent with previous studies and there was no indication that family history, smoking habits and alcohol induced unpleasant side effects have an influence on cortisol secretion.:Inhaltsverzeichnis 3 Abbildungsverzeichnis 6 Tabellenverzeichnis 7 Abkürzungsverzeichnis 8 1 Einleitung 10 1.1 Alkohol – eine kurze Einführung 10 1.2 Bedeutung des Zusammenhangs von HPA-System und Alkohol 12 1.2.1 Experimentelle Alkoholselbstverabreichung 12 1.2.2 Neuropharmakologie von Alkohol 13 1.2.3 Geschlechtsunterschiede bei der Cortisolsekretion 14 1.2.4 Übelkeit und die Auswirkungen auf die Cortisolsekretion unter Alkoholexposition 15 1.3 Orale Alkoholverabreichung versus intravenöse Alkoholverabreichung 16 1.4 Zielsetzung 19 2 Material und Methoden 20 2.1 Versuchsteilnehmer 20 2.1.1 Einschlusskriterien 20 2.1.2 Ausschlusskriterien 21 2.1.3 Aufnahmeuntersuchung 22 2.2 Versuchsaufbau und Durchführung 22 2.2.1 Versuchsprinzip 23 2.2.2 Versuchsablauf 24 2.2.3 Verwendete Materialien 27 2.2.4 CASE Software 27 2.2.5 Herstellung der 6%igen Alkohol-Infusionslösung 29 2.2.6 Messung der BAK 30 2.2.7 Verwendete Fragebögen und Selbsteinschätzungstests 30 2.3 Blutverarbeitung 31 2.3.1 Blutbehandlung im Labor (Testkit der Firma IBL International GMBH) 32 2.3.1.1 Testprinzip im Labor 32 2.3.1.2 Testdurchführung bei Serumproben 32 2.3.1.3 Testauswertung 33 2.4 Statistische Auswertung 33 3 Ergebnisse 37 3.1 Versuchsteilnehmer 37 3.2 Konfirmatorische Datenanalyse der Cortisolsekretion 38 3.2.1 CASE Ergebnisvariablen 38 3.2.2 Analyse von Baseline Cortisol Tag eins vs. Tag zwei 39 3.2.3 Einfluss von Familienanamnese, Geschlecht und maximalem Blutalkohol auf die Cortisolsekretion 40 3.2.3.1 Beobachtung beider Experimentaltage zusammen 40 3.2.3.2 Getrennte Beobachtung für den ersten Experimentaltag 44 3.2.3.3 Getrennte Beobachtung für den zweiten Experimentaltag 46 3.2.3.4 Einfluss von max BAK Tag zwei auf den prozentualen Anteil der Nettofläche an der Gesamtfläche Tag zwei 48 Explorative Analyse von potentiellen weiteren Einflussfaktoren 49 3.3.1 Der Effekt von Übelkeit auf die Cortisolsekretion 49 3.3.2 Der Effekt von Rauchen auf die Cortisolsekretion 50 4 Diskussion 52 4.1 Auswahl der CASE Ergebnisvariablen 52 4.2 Unterschiede zwischen dem ersten und zweiten Experimentaltag 53 4.3 Verschiedene Einflüsse auf die Cortisolsekretion 54 4.3.1 Einfluss von Geschlecht 54 4.3.2 Einfluss von max BAK 55 4.3.3 Einfluss von Familienanamnese 55 4.4 Unterschiede zu vorhergehenden Studien 56 4.5 Diskussion der explorativen Datenanalyse 57 4.5.1 Einfluss von Übelkeit auf die Cortisolsekretion 57 4.5.2 Einfluss von Rauchen auf die Cortisolsekretion 58 4.6 Limitation der D-LAYA Studie 59 4.7 Ausblick 60 Zusammenfassung 61

Les fumeurs consomment des cigarettes de façon intermittente, inhalant chaque cigarette par intermittence, et consommant aussi des cigarettes par intermittence durant la journée. Toutefois, les modèles animaux utilisés pour étudier les effets de la nicotine impliquent un accès continu à la nicotine durant chaque session d’auto-administration. Ici, nous avons donné à des rats un accès intermittent versus continu à la nicotine durant des sessions d’auto-administration. Nous avons ensuite comparé les groupes sur des comportements de consommation caractéristiques de la toxicomanie. Des rats mâles se sont auto-administrés de la nicotine (0.015 ou 0.03 mg/kg/infusion) durant 14 sessions (7h/session). Un groupe avait un accès continu à la nicotine durant chaque session (Long Access, LgA). L’autre groupe avait un accès intermittent (IntA; 12 minutes d’accès à la nicotine par heure). Nous avions donc 4 groupes; LgA-0.015, LgA-0.03, IntA-0.015 et IntA-0.03. Nous avons mesuré la consommation de nicotine et la motivation à consommer différentes doses de nicotine (0.0075, 0.015, 0.03, 0.06mg/kg/infusion), tel que mesuré par l’auto-administration de nicotine sous un horaire de ratio progressif. Après ~15–38 jours d’abstinence, nous avons aussi mesuré la rechute à la consommation induite par une réexposition soit à des stimuli distincts associés à la nicotine, soit à la nicotine elle-même (0.075, 0.15, 0.3 mg/kg, sous-cutané) ou du salin par injection sous-cutanée. Durant les 14 sessions d’auto-administration, les rats LgA-0.03 ont consommé le plus de nicotine. Cependant, après ces 14 sessions, les groupes IntA ont démontré autant de motivation à obtenir la nicotine que les groupes LgA. Les deux groupes avaient aussi une vulnérabilité équivalente à la rechute après la période d’abstinence. L’auto-administration de nicotine par intermittence chez le rat est davantage analogue au mode de consommation de la drogue chez l’humain. De plus, les rats consommant de la nicotine par intermittence prennent nettement moins de drogue que les rats consommant en continu, mais les deux procédures sont tout aussi efficaces pour produire des signes d’addiction à la nicotine. Ceci a des implications pour le raffinement des modèles animaux utilisés dans l’étude des changements neurobiologiques, psychologiques et comportementaux impliqués dans le tabagisme. / Although cigarette smokers consume nicotine and other tobacco products found in cigarettes intermittently throughout each day, procedures to study nicotine effects in laboratory animals involve continuous nicotine access during self-administration sessions. Here, we compared the effects of intermittent versus continuous nicotine self-administration on behavioural features relevant to drug addiction, using an animal model. Four groups of male rats self-administered different doses of nicotine (i.e., 0.015 or 0.03 mg/kg/infusion) during 14 sessions (7 h/session) either as continuous nicotine access (Long Access, or LgA) or intermittent access (IntA; 12 minutes of access to nicotine per hour). We assessed group differences in nicotine intake, as well as nicotine seeking under a progressive ratio schedule of reinforcement (a measure of incentive motivation for drugs). Following nicotine withdrawal, we also measured the susceptibility to cue (withdrawal days 15-30) as well as nicotine-primed (0.075–0.15–3 mg/kg, subcutaneous; withdrawal days 30–38) reinstatement of nicotine-seeking behaviour (measures of relapse). LgA-0.03 rats consumed the most nicotine across the 14 self-administration sessions; however, despite taking much less nicotine than LgA groups, IntA groups showed similar levels of responding for nicotine under a progressive ratio schedule. After nicotine withdrawal, all groups also showed equivalent levels of cue and nicotine induced reinstatement behaviour. IntA nicotine self-administration results in much less drug intake than LgA self-administration, yet the two procedures are equally effective in producing behavioural signs relevant to tobacco addiction. Also, IntA nicotine self-administration serves as a better model of the intermittency of cigarette smoking in humans. Our findings have important implications for preclinical studies investigating the behavioural, psychological, and neurobiological processes involved in tobacco addiction.

Rationale. Adolescence may be a period of susceptibility to the effects of drugs of abuse. This vulnerability may result from a convergence of psychological processes that contribute to drug addiction including impulsive action and incentive motivation during adolescence. Objectives. I examined age differences in incentive motivation, as measured by responding for a conditioned reinforcer (CR) previously paired with natural or drug rewards, and age and sex differences in impulsive action, as measured by responding on a differential reinforcement of low rates of responding (DRL) schedule or premature responding on the 2-Choice Serial Reaction Time Test (2-CSRTT), in Sprague-Dawley rats. The effects of drugs that affect these behaviours in adulthood and that act on neurochemical systems still developing during adolescence were also examined. Methods. In a first set of experiments (Chapter 3), I compared male adolescents and adults on responding for a CR previously paired with sucrose and the effect of amphetamine on this behaviour. In a second set of experiments (Chapter 4), I examined age differences in responding for a CR previously paired with passive or self-administered intravenous (IV) nicotine infusions. Subsequently, I investigated age and sex differences in responding on a DRL schedule in response to amphetamine (Chapter 5) and 2-CSRTT performance in response to amphetamine, nicotine and Ro 63-1908 (a glutamate N-Methyl-D-aspartic acid [NMDA] receptor subunit antagonist; Chapter 6). Results. Compared to adults, adolescents responded more for a CR previously paired with sucrose or passive, but not self-administered, IV nicotine infusions. Amphetamine only enhanced responding for a CR previously paired with sucrose. Adolescents responded more than adults on a DRL schedule, while adolescents made the most premature responses in the 2-CSRTT. No consistent sex differences were observed during the acquisition of either behaviour. Amphetamine increased premature responding most in adolescent males and adult females in the 2-CSRTT but not in responding on the DRL schedule. No consistent age or sex differences were observed for Ro 63-1908 or nicotine. Conclusions. Adolescents show enhanced impulsivity and incentive motivation than adults depending on the behavioural measure. Dopamine may contribute to age and sex differences in these behaviours.

No description available.

Estudos reportando que a autoadministração repetida de drogas de abuso causa mudanças comportamentais, e na expressão de FosB, diferentes às causadas pela administração passiva repetida da mesma droga, em conjunto com estudos de discriminação de estímulos, têm sido chaves para compreender a dependência às drogas. Neste estudo se apresentam resultados de 3 experimentos que avaliaram diferenças gênicas e comportamentais entre a autoadministração de morfina e cocaína sob uma contingência operante, e a administração passiva destas drogas sob uma contingência Pavloviana, usando um modelo de administração de drogas acoplado e um protocolo de transferência operante-Pavloviana (PIT) seletiva em ratos. Os sujeitos foram distribuídos em três grupos: Administração por Contingência Operante (CO), Administração por Contingência Pavloviana (CP) e Controle (Ctr). No Experimento 1, cada sujeito do grupo CO foi exposto a sessões de autoadministração endovenosa de morfina. Depois, a expressão do gene FosB foi medida utilizando uma técnica imuno-histoquímicas em diferentes áreas do cérebro. No Experimento 2 os ratos foram expostos a um protocolo de PIT, treinando de forma inicial as contingências operante e Pavloviana separadamente, em associação a S1, utilizando infusões de morfina como reforçador. Em seguida foi treinado um encadeado de respostas (busca e administração) e finalmente, os sujeitos foram testados para avaliar o controle de estímulos que S1 adquiriu sobre as respostas de busca e administração. O Experimento 3 foi realizado utilizando os métodos dos primeiros dois experimentos, utilizando cocaína como reforçador. Em conjunto, os dados imunohistoquímicos e comportamentais sugerem que a maior expressão de FosB em subáreas envolvidas na dependência às drogas, em comparação entre os grupos CO e CP, está relacionada ao controle de estímulos estabelecido por S1 pelas diferentes contingências de aprendizagem. Ainda, os resultados apontam que estas áreas em que se encontrou uma expressão de FosB diferencial por diferentes contingências de administração de drogas coincidem com algumas das reportadas como envolvidas na PIT. Os resultados estão em concordância com estudos que reportam que a administração repetida de uma droga em contingências operantes ou pavlovianas alteram diferencialmente estruturas cerebrais envolvidas nos processos da dependência às drogas e apoiam a literatura que reporta que o estabelecimento de controle de estímulos que caracteriza a dependência se pode estabelecer por processos de aprendizagem na contingência operante e Pavloviana / Studies reporting that repeated drug self-administration produces behavioral changes, and in FosB expression, different from those produced by repeated passive administration of the same drug have been very important, together with stimulus control studies, have been the key to understand mechanisms underlying drug abuse. This study presents results from 3 experiments evaluating gene and behavioral differences between self-administration of morphine and cocaine under an operant contingency, and passive administration of these drugs under a Pavlovian contingency, using a yoked drug administration model and a selective Pavlovian to instrumental transfer (PIT) protocol in rats. Subjects were divided into three groups: Operant Contingency Administration (CO), Pavlovian Contingency Administration (CP) and Control (Ctr). In Experiment 1, each subject in the CO group was exposed to intravenous morphine self-administration sessions. Then, expression of FosB gene was measured using an immunohistochemical technique in different areas of the brain. In Experiment 2 rats were exposed to a PIT protocol, initially training the operant and Pavlovian contingencies separately in association with S1, using morphine infusions as a reinforcer. Then a chain of responses (seeking and taking) was trained and finally, subjects were tested to evaluate S1 stimulus control over search and administration responses. Experiment 3 was performed using the methods from the first two experiments, using cocaine as a reinforcer. Together, immunohistochemical and behavioral data interact and suggest that a higher expression on FosB expression in subareas involved in drug dependence, in comparison between CO and CP groups, is related to stimuli control established by S1 through the different learning contingencies. Moreover, results point out these same areas in which different FosB expression was found by different drug administration contingencies match some of those reported as being involved in PIT. Results are in agreement with studies reporting that repeated administration of a drug in operant or pavlovian contingencies differentially alter brain structures involved in drug dependence processes and support literature reporting the establishment of stimulus control characterizing addiction can be establish by learning processes in the operant and Pavlovian contingencies

Pesquisas com drogas de abuso têm mostrado consistentemente que a apresentação de estímulos ambientais associados ao uso destas substâncias pode induzir comportamentos de busca e autoadministração das mesmas. Existe a hipótese de que o controle que estímulos ambientais adquirem sobre comportamentos de autoadministração e busca por drogas de abuso poderia ser influenciado tanto pela forma de administração da droga, autoadministração (administração ativa) ou heteroadministração (administração passiva), quanto pela contingência (operante ou respondente) em que a droga foi associada com tais estímulos. Foram formados trios compostos por um sujeito de cada grupo (CONT, ACOP e VEÍC) e realizados dois experimentos. Inicialmente os sujeitos de cada trio foram acoplados por meio de caixas experimentais separadas e expostos a tentativas discretas de apresentação dos estímulos luminosos, S1 e S2, simultaneamente. Como consequência do girar a roda operante na presença de S1 por um integrante do grupo CONT, este recebia uma infusão endovenosa de morfina (0,75 mg/kg), e simultaneamente os animais acoplados no trio recebiam uma infusão de morfina na mesma dose (grupo ACOP) ou de veículo (grupo VEÍC). Posteriormente, os sujeitos de todos os grupos foram treinados a pressionar uma barra por infusões endovenosas de morfina, sem contingência discriminativa programada alguma. Para esta fase, no experimento 1, S2 esteve presente durante as sessões experimentais, porém no experimento 2, nenhum S foi apresentado. Finalmente, os estímulos S1 e S2 foram apresentados em tentativas discretas, em condições de extinção. Os resultados mostram que, durante a extinção, o desempenho dos animais do grupo CONT, mas não os dos grupos ACOP e VEÍC, foi condizente com o treino recebido inicialmente (com 80% ou mais de respostas na barra em presença de S1), indicando que foi estabelecido controle discriminativo sobre a autoadministração de morfina no treino sob a contingência operante, mas não sob a respondente. Estes resultados sugerem que comportamentos de autoadministração e busca por drogas de abuso são influenciados tanto pela forma de administração da droga, quanto pelo tipo de contingência em que uma droga é associada com estímulos ambientais / Drug abuse research has consistently shown that presentation of a drug associated with environmental stimuli can induce drug-seeking and drug-administration behaviors. It has been hypothesized that stimuli control over drug-seeking and self-administration behaviors could be influenced by drug administrations nature, self-administration (active administration) or hetero-administration (passive administration), and also influenced by the drug-stimuli association contingency (operant or respondent). Animals were exposed to right jugular vein catheterization procedure. Groups Contingent (C), Yoked (Y) and Toked Saline (YS) were formed randomly after recovery. Yoked triads were formed with one subject from each group and two experiments were executed. Initially a discriminative training (light stimuli S1 and S2 discrete trials) was presented to triads. Each time C S group member turned an instrumental wheel in presence of S1, simultaneously, an intravenous morphine infusion (0.75 mg/kg) were administrated to that subject and yoked Y group member, as well as an intravenous saline infusion was administrated to yoked YS group member, in every triad. Afterward all subjects were individually trained to lever-press for an intravenous morphine infusion with no discrimination contingency programed. In this phase, S2 was presented through sessions in Experiment 1, while in Experiment 2 no S were presented. After achieving response stability, subjects were exposed to extinction sessions. Stimuli S1 and S2 discrete trials were presented but no consequences were programed for bar-press responses. Results show that only C groups performance was consistent with the previously received discriminative training (80% or more of bar-press responses in S1 presence) during extinction sessions, indicating that an operant training, but not a respondent training, successfully established morphine self-administration discriminative control. These results suggest that self-administration behaviors are in fact influenced by drugs administration nature and also by the drug-stimuli association contingency