Psychostimulant-induced reinstatement of place preferences have been used to investigate underlying physiological mechanisms mediating drug-seeking behavior in adolescent and adult rodents; however, it is still unclear how psychostimulant exposure during adolescence affects neuronal communication in the mesolimbic dopamine (DA) pathway and whether these changes would elicit enhanced drug-seeking behavior later in adulthood. The aim of the present study was to investigate the effects of intra-ventral tegmental area (VTA) or intra-nucleus accumbens septi (NAcc) DA D2 receptor antagonist infusions on cocaine-induced reinstatement of cocaine place conditioning in high and low responders for cocaine reward. Adolescent rats were exposed to cocaine place conditioning [postnatal day (PND 28-39)] and divided into high and low responders for cocaine reward based on their place preference expression score. Place preferences were extinguished and guide cannula were implanted into either the VTA or NAcc followed by one of the following: 1) intra-VTA or intra-NAcc infusion of the DA D2 receptor antagonist sulpiride (100 µM) during a cocaine-primed reinstatement test (10 mg/kg/ip cocaine) or 2) measurement of NAcc DA levels during intra-VTA or intra-NAcc infusion of sulpiride (100 µM), a cocaine prime (10 mg/kg cocaine) and re-exposure to the cocaine paired chamber. Infusion of sulpiride into the VTA but not the NAcc blocked reinstatement of cocaine place conditioning in rats exposed to cocaine during adolescence. Furthermore, re-exposure to cocaine-associated cues and simultaneous local infusion of sulpiride into either the VTA or NAcc attenuated cocaine-induced increases in accumbal DA levels for rats pretreated with cocaine during adolescence, regardless of phenotype. These data suggest intrinsic compensatory mechanisms in the mesolimbic DA pathway mediate adolescent behavioral responsivity to cocaine prime-induced reinstatement of cocaine place conditioning later on in adulthood.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 82-94).

<p>Activation of monoaminergic receptors is essential to the mechanism by which psychostimulants and antidepressants induce changes in behavior. Although these drugs rapidly increase monoaminergic transmission, they need to be administered for several weeks or months in order to produce long-lasting alterations in behavior. This observation suggests that it is likely that molecular mechanisms downstream of receptor activation contribute to the effects of psychostimulants and antidepressants on behavior. </p><p>Recently, we and others have demonstrated that the methyl-CpG-binding protein 2 (MeCP2) contributes to both neural and behavioral adaptations induced by repeated psychostimulant exposure (Deng et al, 2010, Im et al, 2010). Psychostimulants induce rapid and robust phosphorylation of MeCP2 at Ser421 (pMeCP2), a site that is thought to modulate MeCP2-dependent chromatin regulation (Cohen et al, 2011), and this phosphorylation event is selectively induced in the GABAergic interneurons of the nucleus accumbens (NAc). In order to understand the signaling pathways that contribute to the pattern of pMeCP2 we observe, I characterized the monoaminergic signaling pathways that regulate pMeCP2. I found that activation of dopamine (DA) and serotonin (5-HT) transmission is sufficient to induce pMeCP2. The novel finding that drugs that activate serotonergic signaling induce pMeCP2 suggests that pMeCP2 may be involved in serotonergic mediated behaviors.</p><p>To determine the requirement of pMeCP2 in serotonergic mediated behaviors, I utilized mice that bear a knockin (KI) mutation that converts serine to alanine at 421 (S421A) (Cohen et al, 2011). After characterizing the behavioral phenotype of these mice, I conducted tests to assess anxiety- and depression-like behavior. I found that the KI mice do not display heightened anxiety in several assays. However, the KI mice exhibit depression-like behavior in the forced swim and tail suspension but show no differences compared to wild-type (WT) littermates in the sucrose preference test, suggesting that pMeCP2 may be implicated in the behavioral response to stressful stimuli. </p><p>Because we are interested in examining the role of pMeCP2 in the behavioral adaptations to chronic monoaminergic signaling, I then put the KI mice and their WT littermates through chronic social defeat stress, a behavioral paradigm in which repeated exposure to aggressive mice causes social avoidance that is reversed by chronic but not acute antidepressant treatment. Although the WT mice show an increase in social interaction following chronic imipramine treatment, the KI mice fail to show a behavioral response to chronic treatment. These data suggest that pMeCP2 may be implicated in the antidepressant action of chronic imipramine. Finally, investigation of the brain regions in which pMeCP2 may be contributing to the behavioral response to chronic imipramine treatment revealed that chronic but not acute imipramine treatment induces pMeCP2 in the lateral habenula (LHb), a brain region involved in the behavioral response to stress and reward. Together, these data implicate a novel role for pMeCP2 in depression-like behavior and the behavioral response to chronic antidepressant treatment.</p> / Dissertation

In humans, adolescent exposure to illicit drugs predicts the onset of adult drug abuse and suggests that early drug use potentiates adolescent vulnerability to drug addiction. In experiment 1, it was hypothesized that adolescent rats would show a CPP for a low cocaine dose if in fact adolescents are more vulnerable to cocaine's rewarding effects. Place preferences were measured in early adolescent [postnatal day (PND) 35], late adolescent (PND 45) and young adult (PND 60) rats by injecting either 0, 5 or 20 mg/kg cocaine and conditioning them to environmental cues in a 2-chamber place conditioning apparatus. Significant cocaine preferences were found for all ages at the high dose. Interestingly, PND 35's were the only age group to have a CPP at the low dose suggesting that PND 35 rats are more sensitive than late adolescent and young adult rats to cocaine's rewarding effects. In Experiment 2, it was hypothesized that age-related differences in cocaine CPP may be mediated by differences in the mesolimbic dopaminergic (DA) system throughout development. Extracellular DA levels in the nucleus accumbens septi (NAcc) of early adolescent, late adolescent and adult rats were measured via quantitative microdialysis. PND 35, PND 45 and PND 60 rats were injected daily with either 5 mg/kg/ip or saline for 4 days, surgically implanted with a microdialysis probe aimed at the NAcc. Rats were perfused with either 0, 1, 10 or 40 nM DA and the extracellular DA concentration was measured. Our results show that adolescents differ from adults in basal DA with PND 35 rats having low basal DA (0.4 nM), PND 45 rats having high basal DA (1.8 nM) and PND 60 rats having intermediate basal DA (1.3 nM). PND 45 cocaine treated rats showed a 58% decrease in basal DA. All cocaine treated rats, regardless of age, showed a significant increase in DA over baseline in response to a cocaine challenge. Additionally, there were age-related differences in the extraction fraction (Ed), an indirect measure of DA reuptake, with PND 45 and PND 60's showing a decrease in basal Ed, an effect absent in PND 35's. Together these findings suggest that there are substantial ontogenetic differences in extracellular DA and DA reuptake and that these differences may provide an explanation for adolescent vulnerability to addiction. Future research should investigate DA supply and degradation processes in naïve and cocaine treated adolescent rats and vulnerability to addiction.

The long-term consequences of adolescent drug use in shaping a network primed for addiction is an issue of utmost importance. The use of cocaine during adolescent development could alter the normal growth of the reward system and affect the adult mesolimbic system, however, there is scant literature aimed at finding out if animals are more vulnerable to the adverse effects of drugs during adolescence. The present study investigated whether cocaine pretreatment in adolescent and adult rats produced differences in cocaine-induced neurochemical cross-sensitization to a naturally reinforcing substance in adulthood. To evaluate the responsivity of the mesolimbic system after repeated cocaine, sucrose was offered during the dialysis procedure and dialysate was collected. All saline pretreated rats had significant increases in DA levels compared to baseline levels and there were no difference in the age of pretreatment. Rats pretreated with cocaine as adults also had significant increases in DA levels after sucrose. Interestingly, sucrose intake significantly enhanced DA levels in cocaine pretreated adolescent rats. The results from this experiment clearly show that in rats pretreated with cocaine during adolescence there is an enhance response of the DAergic system in response to a naturally reinforcing substance therefore; cocaine exposure during adolescence results in persistent long term changes in the mesolimbic pathway. Future studies need to ascertain the underlying mechanisms and their role in the process of addiction.

Dopamine increases in the nucleus accumbens after contingent and noncontingent ethanol administration in rats, but the contributions of the core, coreshell border, and shell subregions to this response are unclear. Also, it is not fully understood if increases in dopamine under these circumstances are due to the pharmacological effects of ethanol, stimuli associated with administration, or both. The studies presented in this dissertation were conducted to investigate dopamine’s role in each of these accumbal regions during ethanol administration and presentation of associated stimuli. Using microdialysis, ethanol and dopamine concentrations in accumbal subregions were measured every five minutes before, during, and after either experimenter-delivered intravenous ethanol or operant ethanol self-administration. After intravenous ethanol infusions, the increase in dopamine in the shell of the accumbens was significantly higher than that observed in the core. During operant ethanol self-administration, the core, core-shell border, and shell, all exhibited significant increases in dopamine during transfer of the animal into the operant chamber, with animals trained to drink sucrose + ethanol showing significantly higher increases when compared to those trained to drink sucrose alone. Dopamine increased significantly only in the core-shell border during ethanol consumption, and dopamine levels in the core and shell responded in a similar manner during all phases of the experiment. Together, these results suggest that dopamine responses to intravenous ethanol infusions and operant ethanol self-administration are subregion specific. Also, while increases in dopamine resulting from intravenous ethanol infusions in naïve animals appear to be due to the pharmacological effects of the drug, increases in ethanol-experienced animals during transfer into the operant chamber, and during ethanol consumption, may also be due to stimuli associated with ethanol administration. / text

Mesolimbic dopamine plays an important role in ethanol reinforcement, and studies have shown that accumbal dopamine increases during operant ethanol self-administration. However, no one has ever studied this dopaminergic response during the acquisition of ethanol self-administration. Furthermore, some studies have shown that the dopamine signal does not correlate with the pharmacological effects of ethanol, but with the time during which the animal consumes the majority of the ethanol solution and when the sensory stimuli of ethanol are strongest. However, there is currently no direct evidence showing that the sensory stimuli of ethanol is indeed what causes the brief increase in accumbal dopamine during ethanol self-administration. The studies in this dissertation attempted to elucidate these issues. We designed and tested a placebo spout, which was to be used to study the relationship between accumbal dopamine and the sensory stimuli of ethanol during self-administration. Unfortunately, the placebo designs were either not feasible for performing microdialysis or did not show promising behavioral data. We also developed and tested a self-administration protocol in which the concentrations of ethanol (10%) were kept constant throughout the study. The new protocol was successful in initiating and maintaining ethanol self-administration, and the animals doubled their intake from day 1 to day 2 of ethanol consumption. Using this protocol, we trained male Long Evans rats to self-administer ethanol and measured accumbal dopamine during the first two days of ethanol self-administration through microdialysis. The behavioral and neurochemical data matched. A single exposure to ethanol was sufficient for the animals to double their ethanol consumption by day 2 and to cause an increase in accumbal dopamine during the first 5 minutes of ethanol self-administration. The dopamine response was observed during the time when the sensory stimuli of ethanol were strongest, but before ethanol reached peak concentrations in the brain. Overall, these results suggest that the dopamine response to ethanol self-administration may not be solely pharmacological and that a single exposure to ethanol is sufficient to learn the association between ethanol and its cues. These findings give us greater insight into mesolimbic dopamine's role in the early stages of ethanol reinforcement. / text

Pleasure is part of hedonic well-being, with roots back to Epicurus 2000 years ago. With the new evolving neuroscientific methods of the late 20th and beginning of the 21st century, we are now able to study the biological components of pleasure. This thesis aims to review empirical studies on the neural correlates of pleasure, which can have important implications for well-being, and treatment of addiction and affective disorders. Recent studies have suggested that pleasure can be separated into coding and causing. Discoveries show that causing of pleasure is created in so called hedonic hot spots, areas of the brain that intensely creates pleasure in the shell of nucleus accumbens and in the ventral pallidum. Areas that codes pleasure on the other hand is represented into more cortical areas of the brain, including orbitofrontal cortex, anterior cingulate cortex and anterior insular cortex. There has been a growing understanding about how pleasure is represented in the brain, and a discussion on interpretations and limitations are provided followed by future research suggestions in the final section.

Thesis (Ph. D.)--University of Texas at Austin, 2006. / Vita. Includes bibliographical references.

Thesis (Ph. D.)--Cornell University, May, 2008. / Vita. Includes bibliographical references (leaves 143-165).