A monitorização terapêutica tem sido descrita como valioso recurso clínico, na individualização do regime de dosagem, de acordo com a concentração do fármaco e/ou de seus produtos de biotransformação, em amostras de plasma ou soro, coletadas com base no contexto clínico e nos princípios da farmacocinética. Em razão da complexidade dos fluidos biológicos e da baixa concentração dos fármacos nestas matrizes, a etapa de preparo de amostra, extração, pré-concentração dos analitos e eliminação dos interferentes, têm sido requerida para o desenvolvimento de métodos cromatográficos com alta sensibilidade e seletividade analítica. A extração sortiva em barra de agitação (SBSE), recente técnica de preparo de amostra, baseia-se no equilíbrio de sorção do analito entre as fases: extratora (polidimetilsiloxano) e amostra aquosa. A fase extratora PDMS é a única disponível no mercado, o que tem limitado a sensibilidade e a seletividade analítica da técnica SBSE. O uso de polipirrol (PPY) como fase extratora está relacionado às diferentes interações de seus grupos funcionais (hidrofóbica, -, com o grupo funcional polar, troca iônica, ácido-básica, dipolo-dipolo e dipolo induzido-dipolo.) com os analitos. Neste trabalho, uma nova fase extratora SBSE, com o revestimento misto PDMS/PPY foi avaliada para análises de antidepressivos em amostras de plasma, por cromatografia líquida de alta eficiência. A otimização das variáveis SBSE: tempo e temperatura de extração, tempo de dessorção e pH da matriz biológica, baseada no equilíbrio de sorção dos analitos entre as fases: polimérica (PDMS/PPY) e fluido biológico, permitiu a determinação dos analitos em concentrações plasmáticas que contemplam o intervalo terapêutico. A presença de estrutura porosa (PPY) e não porosa (PDMS) na superfície polimérica da barra extratora SBSE-PDMS/PPY foi confirmada através de análises por Microscopia Eletrônica de Varredura (MEV). De acordo, com os resultados obtidos nas análises de MEV e nas análises (individuais e simultâneas) de amostras de plasma enriquecidas com os analitos, os mecanismos de retenção dos fármacos junto à superfície PDMS/PPY ocorreram através dos processos de adsorção (PPY) e absorção (PDMS). A validação analítica foi realizada segundo as normas da Agência Nacional de Vigilância Sanitária. O método padronizado apresentou linearidade no intervalo de concentração plasmática que variou dos limites de quantificação a 500 ng mL-1, coeficientes de determinação maiores que 0,994, precisão inter ensaios com coeficientes de variação menores que 15% e exatidão de 96% a 106%. Os valores de limites de quantificação obtidos são congruentes com a menor concentração plasmática do intervalo terapêutico preconizado. O método SBSE-PDMS/PPY padronizado e validado foi utilizado para determinações dos antidepressivos, sertralina, duloxetina e fluoxetina em amostras de plasma de pacientes, em terapia com estes fármacos. Desta forma, o método SBSE-PDMS/PPY poderá ser empregado para fins de monitorização terapêutica. / Therapeutic drug monitoring has been described a valuable clinical resource for the customization of the dosage regimen, in accordance with the drug concentration and/or its biotransformation products, in of plasma or serum samples, collected on the basis of clinical context and pharmacokinetics principles. Due the complexity of biological fluids and the low concentration of the drug in these matrices, an stage of sample preparation, extraction, pre-concentration of the analytes and elimination of the interferents has been required for the development of chromatographic methods with high sensitivity and analytical selectivity. Stir bar sorptive extraction (SBSE), a recent sample preparation technique, is based on the sorption equilibrium of the analytes between the polydimethylsiloxane (PDMS) and aqueous phases. PDMS is the only commercially available extraction phase (SBSE), which has limited the analytical sensitivity and the selectivity of the SBSE technique. The use of polypyrrole (PPY) as extraction phase is related to the different interactions of its functional groups (- interactions, polar groups interactions, acid-base, dipole-dipole, dipole-induced-dipole) and the analytes. In this work, a new SBSE extraction phase, with a PDMS/PPY coating was evaluated for the analysis of antidepressants in plasma samples by High Performance Liquid Chromatography. The optimization of the SBSE variables extraction time and temperature, dessorption time and pH of the biological matrix based on the sorption equilibrium of the analytes between the polymeric (PDMS/PPY) and biological fluid phases, allowed determination of the analytes in plasmatic concentrations that correspond to the therapeutic interval. The presence of a porous structure (PPY) as well as no none, porous (PDMS), on the polymeric surface of SBSE-PDMS/PPY was confirmed by Scanning Electron Microscopy (SEM) analyses. In agreement with the results obtained by SEM analyses and individual and simultaneous analyses of the plasma samples spiked with the analytes, the mechanisms of drugs retention on the surface of PDMS/PPY occur through adsorption (PPY) and absorption (PDMS). Analytical validation was carried through according to the norms of the National Agency of Sanitary Vigilance. The standardized method presented linearity in the plasmatic interval concentrations that varied from the limits of quantification to 500 ng mL-1, the determination coefficients were higher than 0.994, inter precision assays with coefficients of variation lower than 15%, and accuracy from 96% to 106%. The quantification value limits were in agreement with the lowest plasmatic concentration of the established therapeutical interval. The standardized and validated SBSE-PDMS/PPY method was used for determination of sertraline, duloxetine and fluoxetine in plasma patient samples under therapy with these drugs. Thus, the SBSE-PDMS/PPY method could be used for therapeutic drug monitoring.

A relação entre as alterações estruturais cerebrais e a resposta ao tratamento da depressão em idosos continua a ser uma área intrigante de pesquisa. Neste estudo, foram investigadas diferenças quanto ao volume total e regionalizado de substância cinzenta e branca em exames de ressonância magnética (RM) de idosos com depressão (de acordo com os critérios do DSMIV-TR) e de controles. Além disso, para melhor compreender a fisiopatologia da depressão no idoso, o volume total das hiperintensidades em substância branca foi quantificado e comparado entre os grupos. A amostra foi composta por 30 idosos com depressão e 22 controles. Os idosos com depressão foram divididos em grupos de acordo com o uso prévio de antidepressivos, a resposta ao tratamento farmacológico, assim como de acordo com a idade de início da depressão. As imagens de RM foram processadas utilizando o programa Statistical Parametric Mapping e a morfometria baseada em voxel (DARTEL). A quantificação do volume total das hiperintensidades em substância branca foi realizada através de uma variação do método automático conhecido como Expectation Maximization Segmentation (EMS). Na análise do cérebro inteiro, encontramos uma redução volumétrica significativa no giro reto e no córtex orbitofrontal bilateralmente em pacientes em comparação com os controles.Além disso, os pacientes que não estavam em uso de antidepressivos no momento da aquisição da RM apresentaram uma redução volumétrica ainda maior no giro reto e no córtex orbitofrontal bilateralmente. Pacientes com uso prévio de antidepressivos mostraram uma redução volumétrica no giro reto e córtex orbitofrontal bilateralmente em comparação com os controles e pacientes sem uso prévio de antidepressivos apresentaram uma redução ainda maior no giro reto e no córtex orbitofrontal bilateralmente em comparação com os controles. De acordo com seu estado de remissão após o uso de um algoritmo de tratamento farmacológico da depressão em idosos, os pacientes foram classificados nos grupos remissão e não remissão. O grupo remissão apresentou uma redução volumétrica no giro reto e no córtex orbitofrontal bilateralmente e no pólo temporal médio direito, em comparação com o grupo controle. O grupo não remissão mostrou uma redução volumétrica ainda maior no giro reto e no córtex orbitofrontal bilateralmente em comparação com o grupo controle. Para investigar fatores preditores de remissão, foi utilizada uma regressão logística. A pontuação inicial do Mini Exame do Estado Mental e o volume corrigido (para o volume total de substância branca) inicial do córtex orbitofrontal superior lateral esquerdo classificaram os pacientes de acordo com a resposta ao tratamento farmacológico. As mesmas análises estatísticas foram realizadas em relação ao volume de substância branca. Pacientes sem uso prévio de antidepressivos mostraram uma redução volumétrica no giro frontal superior direito em comparação com os controles. Além disso, o grupo com depressão de início tardio apresentou um aumento volumétrico no lóbulo posterior esquerdo do cerebelo em comparação com o grupo controle. Em relação ao volume total das hiperintensidades de substância branca, a amostra final processada com o EMS foi constituída por 22 pacientes e 19 controles. As mesmas comparações entre grupos descritas anteriormente foram realizadas e não foi encontrada diferença estatisticamente significante entre os grupos. Foi realizada uma nova regressão logística para investigar fatores preditores de remissão nessa amostra menor, incluindo o volume total de hiperintensidades de substância branca e não foi encontrado resultado estatisticamente significante. Em conclusão, os idosos com depressão apresentam redução volumétrica no giro reto e no córtex orbitofrontal bilateralmente e isso pode ser uma característica da depressão em idosos e um potencial biomarcador dessa condição. O uso de antidepressivos pode proteger contra a redução volumétrica de substância cinzenta e branca e parece ter um efeito neurotrófico nesses pacientes. Além disso, déficit cognitivo e redução de substância cinzenta no córtex orbitofrontal superior lateral esquerdo no início do estudo podem ser preditores de pior resposta ao tratamento farmacológico da depressão no idoso. Estudos longitudinais, com amostras maiores e com pacientes com depressão mais grave poderão contribuir para uma melhor compreensão da fisiopatologia da depressão em idosos / The association between structural brain changes and treatment response in patients with late-life depression (LLD) remains an intriguing area of research. This magnetic resonance imaging (MRI) study investigated the baseline gray and white matter volume of elderly with and without major depression (according to the DSM-IV-TR criteria). Moreover, to better understand the pathophysiology of LLD, white matter hyperintensities total volume was quantified and compared among groups. The sample consisted of 30 elderly patients with depression and 22 elderly healthy controls. Depressed patients were classified according to their previous use of antidepressants, to their response to pharmacological treatment, and to their age of depression onset. Brain MRI scans were processed using statistical parametric mapping and voxel-based morphometry (DARTEL). White matter hyperintensities total volume was conducted with a variation of an automatic method known as Expectation Maximization Segmentation (EMS). In the whole-brain analysis, we found a significant volumetric reduction in the gyrus rectus and in the orbitofrontal cortex (OFC) bilaterally in patients in comparison with controls. Additionally, patients who were not taking antidepressants at the time of the MRI had an even greater volumetric decrease in the gyrus rectus and in the orbitofrontal cortex bilaterally. Besides, we classified patients according to their previous antidepressant use (with or without) and compared them with controls. Patients with previous antidepressant use had a volumetric reduction in the gyrus rectus and in the OFC bilaterally in comparison with controls and patients without previous antidepressant use had an even greater reduction in the gyrus rectus and in the OFC bilaterally in comparison with controls. According to their remission status after the use of a pharmacologic algorithm treatment, patients were classified in remitted or not remitted. In comparison with controls, remitted patients showed a volumetric reduction in a cluster that included the gyrus rectus and in the OFC bilaterally and in another cluster that included the right middle temporal pole. Non-remitted patients showed an even greater volumetric reduction in the gyrus rectus and in the OFC bilaterally compared with controls. A logistic regression was used to investigate baseline predictive factors of remission. Baseline Mini Mental State Examination scores and the left superior lateral OFC standardized (to the total gray matter volume) volume classified patients with respect to their remission status. Regarding white matter volume, the same statistical analyses were conducted. Patients without previous antidepressant use showed a volumetric reduction in the right superior frontal gyrus in comparison with controls. In addition, late onset depression group had a volumetric increase in the left posterior cerebellum lobe in comparison with controls. In relation to the white matter hyperintensities total volume, 22 patients and 19 controls were processed with EMS. The same comparisons among groups were conducted and no statistical significant difference was found. A logistic regression was conducted to investigate baseline predictive factors of remission in this smaller sample, including the white matter hyperintensities total volume and no statistically significant result was found. In conclusion elderly with depression have volumetric reduction in the gyrus rectus and in the OFC bilaterally and this may be characteristic of late life depression and a potential biomarker of this condition. Antidepressant use seems to protect against gray matter reduction and to have a neurotrophic effect. In addition, cognitive deficits and regional gray matter abnormalities at baseline seem to be predictors of worse response to antidepressant treatment. Further longitudinal studies with larger sample size and more severe depression intensity may contribute to a better understanding of the pathophysiology of depression in the elderly

Les antidépresseurs sont les traitements pharmacologiques de choix de la dépression. Pourtant devant un nombre non négligeable d’études « négatives » et l’absence d’efficacité dans les dépressions peu sévères, la question de leur utilité reste controversée. Cette controverse présuppose que l’efficacité des antidépresseurs se mesure facilement. Pourtant, comme dans toute expérience, le dispositif expérimental peut influencer le résultat mis en évidence. L’objectif de cette thèse est justement d’explorer l’incidence de la procédure expérimentale sur l'effet mesuré des pharmacothérapies antidépressives. Est-ce que leur efficacité mesurée dans les études randomisées reflète leur effectivité (dans la « vraie vie ») ? Pour répondre à cette question, nous avons conduit une méta-analyse d’un large panel d’études de méthodologies très différentes. Une méta-régression a été réalisée pour modéliser la réponse au traitement en fonction des caractéristiques des études. Elle a permis de mettre en évidence 1/ une réponse supérieure dans les essais randomisés par rapport aux études observationnelles (possiblement plus représentatives de la « vraie vie »); 2/ une variation de la réponse en fonction de la durée de l’étude, du nombre de visites de suivi, de la population de patients inclus, du type d’analyse choisie, de la possibilité d’avoir un placebo et du design en aveugle. Une revue de la littérature apporte ensuite une perspective qualitative à la question posée. L’objectif était de décrire les dispositifs expérimentaux actuellement utilisés ou envisageables dans l’évaluation des antidépresseurs. La revue de la littérature 1/ a permis de préciser les limites des études actuelles en termes de validité interne (mesure des critères de jugement, attrition, levée de l’aveugle importance de la réponse placebo) et externe (difficultés majeures de généralisations des études); 2/ conduit à proposer une évaluation multimodale des antidépresseurs dans laquelle les études observationnelles auraient toute leur place. La question de la mesure est ensuite abordée à travers des simulations. Après avoir exploré l’impact du dispositif expérimental au niveau de l’essai en soi, le phénomène est analysé à un niveau plus global. Le choix a été fait d’un raisonnement par l’absurde, sur le terrain de la « comparative effectiveness ». L’objectif était de comparer les placebos d’antidépresseurs grâce à une méta-analyse en réseau. Ces comparaisons indirectes n’ont pas permis de mettre en évidence de différence entre placebos d’antidépresseurs. Un biais de publication a pu être mis en évidence et empêche de conclure formellement sur un résultat pourtant évident. Ce résultat ouvre une discussion sur le degré de confiance que l’on peut apporter à ce que l’on considère généralement comme des preuves scientifiques de l’effectivité de nos pratiques. Nos résultats impliquent un nouvel éclairage à la controverse sur les antidépresseurs en replaçant le débat sur le terrain épistémologique. / Antidepressant drugs have become the most prescribed and used pharmacological treatment for depressive disorders. Yet, their usefulness is still being debated in view of a significant number of “negative” studies and of their lack of efficacy in mild depressive disorders. This controversy presupposes that the efficacy of antidepressants can easily be measured. However, as in any experiment, the experimental method may influence the outcome. The aim of this thesis is precisely to explore the impact of the experimental procedures on the measured effect of antidepressant therapies. Also we ask the question of whether their efficacy measured in randomized trials reflects their “effectiveness” (in a “real life” setting)? To answer these questions, we performed a meta-analysis of a large number of studies with different methodologies. A meta-regression was performed to modelize the response to the treatment according to the studies’ characteristics. It highlighted 1/ a superior response in randomized trials compared to observational studies (possibly more representative of the “real life”); 2/ a variation of the response according to the duration of the study, to the number of follow-up visits, to the type of included patients, to the type of analysis, to the use or not of a placebo and to the use or not of a blind design. A review of the literature provides a qualitative prospect to the question. The objective was to describe the experimental designs used in the assessment of antidepressants and to identify possible innovations. The review of the literature 1/ has permitted to state precisely the limits of the studies in terms of internal validity (measurement of assessment criterions, attrition, unblinding, importance of the placebo response) and external validity (major difficulties with generalization of the studies’ results); 2/ leads to offer a multimodal evaluation of antidepressants in which observational studies would have their own place. Then the issue of measurement is specifically approached, using simulations. After exploring the impact of the experimental procedure at the level of the trial itself, the phenomenon is explored at a more global level. The choice of a reductio ad absurdum on the field of comparative effectiveness was decided. The aim was to compare the placebos of different antidepressants in a network meta-analysis. Indirect comparisons did not show any difference between placebos of antidepressants but a significant publication bias was detected. It does not allow drawing any firm conclusion on a result which is still obvious. This result invites discussion as to whether the evidence that we usually rely on provides us with a reasonable degree of certainty about the nature and effectiveness of our practices. Our results entail a new light on the controversy about the efficacy of antidepressants by replacing the debate on the epistemological ground.

It has been reported that locus coeruleus (LC) degeneration precedes the degeneration of other neurons in the brain in some neurodegenerative diseases like Alzheimer’s disease (AD) and Parkinson’s disease (PD). However, the precise mechanisms of neurodegeneration remain to be elucidated. N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) has been widely used as a noradrenergic neurotoxin in the development of AD and PD animal models for specific LC degeneration. However, the precise mechanism of action of DSP4 remains unclear. An increased systemic DNA damage caused by neurotoxin or oxidative stress has been found to be related to the pathogenic development of neurodegeneration. The process of neurodegeneration is not well understood, so current therapeutic approaches are limited to disease management and symptoms relief, such as using antidepressants for depression symptoms, which often accompany neurodegenerative disorders. To date, few studies have explained why different groups of antidepressants have similar clinical effects on relieving depression. Our data demonstrate that DSP4 induces the DNA damage response (DDR) and results in down-regulation of dopamine β-hydroxylase (DBH) and the norepinephrine transporter (NET), which are 2 noradrenergic phenotypes. DSP4 results in cell cycle arrest in S and G2/M phases, which is reversible. The comet assays verify that DSP4 induces single-strand DNA breaks (SSBs). Furthermore, the neurotoxins camptothecin (CPT) and DSP4 were used to induce the DDR in SH-SY5Y cells, fibroblast cells, and primary cultured neurons. Data show that both CPT and DSP4 induce the DDR in SH-SY5Y cells and primary cultured LC neurons. Compared to fibroblast cells, SH-SY5Y cells and LC neurons are more sensitive to the accumulation of DNA damage when treated with CPT or DSP4. Persistent phosphorylated H2AX (γH2AX) and p53 (p-p53ser15) levels indicate a deficient repair in noradrenergic SH-SY5Y cells and LC neurons. In addition, the current study demonstrates that some antidepressants reduce the DDR induced by DSP4 or CPT in SH-SY5Y cells. Flow cytometry data show that selective antidepressants protect cells from being arrested in S-phase. Together, these effects suggest that blocking DNA damage is one important pharmacologic characteristic of antidepressants, which may explain why different antidepressants could alleviate depression symptoms in neurodegenerative patients.

Dementia is a clinical syndrome caused by neurodegeneration or cerebrovascular injury. Patients with dementia suffer from deterioration in memory, thinking, behavior and the ability to perform everyday activities. Since there are no cures or disease-modifying therapies for dementia, there is much interest in identifying modifiable risk factors that may help prevent or slow the progression of cognitive decline. Medications are a common focus of this type of research. Importantly, according to a report from the Centers for Disease Control and Prevention (CDC), 19.1% of the population aged 60 and over report taking antidepressants during 2011-2014, and this number tends to increase. However, antidepressant use among the elderly may be concerning because of the potentially harmful effects on cognition. To assess the impacts of antidepressants on the risk of dementia, we conducted three consecutive projects. In the first project, a retrospective cohort study using Marginal Structural Cox Proportional Hazards regression model with Inverse Probability Weighting (IPW) was conducted to evaluate the average causal effects of different classes of antidepressant on the risk of dementia. Potential causal effects of selective serotonin reuptake inhibitors (SSRIs), serotonin and norepinephrine reuptake inhibitors (SNRIs), atypical anti-depressants (AAs) and tri-cyclic antidepressants (TCAs) on the risk of dementia were observed at the 0.05 significance level. Multiple sensitivity analyses supported these findings. Unmeasured confounding is a threat to the validity of causal inference methods. In evaluating the effects of antidepressants, it is important to consider how common comorbidities of depression, such as sleep disorders, may affect both the exposure to anti-depressants and the onset of cognitive impairment. In this dissertation, sleep apnea and rapid-eye-movement behavior disorder (RBD) were unmeasured and thus uncontrolled confounders for the association between antidepressant use and the risk of dementia. In the second project, a bias factor formula for two binary unmeasured confounders was derived in order to account for these variables. Monte Carlo analysis was implemented to estimate the distribution of the bias factor for each class of antidepressant. The effects of antidepressants on the risk of dementia adjusted for both measured and unmeasured confounders were estimated. Sleep apnea and RBD attenuated the effect estimates for SSRI, SNRI and AA on the risk of dementia. In the third project, to account for potential time-varying confounding and observed time-varying treatment, a multi-state Markov chain with three transient states (normal cognition, mild cognitive impairment (MCI), and impaired but not MCI) and two absorbing states (dementia and death) was performed to estimate the probabilities of moving between finite and mutually exclusive cognitive state. This analysis also allowed participants to recover from mild impairments (i.e., mild cognitive impairment, impaired but not MCI) to normal cognition, and accounted for the competing risk of death prior to dementia. These findings supported the results of the main analysis in the first project.

Abstract The prevalence of diabetes in the United States in 2015 was 30.3 million people; which is the equivalent of 9.4% of the population. It is a major contributor to morbidity and mortality. Depression is often underdiagnosed in these patients, which contributes to poor self-management and poor health outcomes. In a large primary-care practice on the East Coast, there is no guideline for depression screening in the diabetic population. Focusing on this specific primary care setting, the research question addressed the underdiagnoses of depression in diabetic patients. The project evaluated the prevalence of depression among patients diagnosed with diabetes by using the U.S. Preventative Services Task Force (USPSTF) depression screening tool called the Patient Depression Questionnaire. The Grove Model for Implementing Evidence-Based Guidelines in Practice Framework was used in systematically integrating the new practice guideline of screening all diabetes patients for depression. The framework involves identifying the practice problem, locating, and collecting the most well-founded evidence, and evaluating the quality of the evidence-based guideline of screening all diabetes patients for depression. To answer the project question, 135 diabetic patients in a primary care outpatient setting completed the Patient Depression Questionnaire. Descriptive statistics was used to describe the sample and determine the frequency of scores throughout the sample of patients. Ranges of scores and percentages were computed using frequency distribution. Of these 135 diabetic patients, 63.7% showed some level of depression ranging from mild to severe, which was undiagnosed prior to the administration of the screening tool. As a result of the project, recommendations were made to the site to implement the use of the depression screening tool as policy.

<p>Eukaryotic gene transcription plays a regulatory role in mammalian developmental processes. It has been shown that transcriptional control is an important mechanism for specification of neurotransmitter phenotypes. In the mammalian central nervous system, the transcription factor AP-2 family is one of the critical regulatory factors for neural gene expression and neuronal development. It has been shown that several genes in the monoaminergic systems have AP-2 binding sites in regulatory regions, suggesting a regulatory role of AP-2 also in the adult brain. Brainstem monoamines are implicated in the expression of personality traits and imbalances in these systems may give rise to psychiatric disorders. </p><p>The gene encoding AP-2β includes a polymorphic region consisting of a tetranucleotide repeat of [CAAA]_4-5 in intron 2. Studies on AP-2β genotype in relation to personality and platelet MAO activity, a trait-dependant marker for personality, are presented in this thesis. Furthermore, correlations between brainstem levels of AP-2α and AP-2β and monoamine turnover in projection areas in rat forebrain are reported. These results strengthen the notion that the AP-2 family is important regulators of the monoaminergic systems in the adult brain. Furthermore, two studies are presented in this thesis with analyses indicating a role for AP-2 in the molecular mechanism of antidepressant drugs.</p><p>Altogether, this thesis presents data supporting our notion that the transcription factor AP-2 family is involved in the regulation of the monoaminergic systems both pre- and postnatally, and, therefore, might be involved in the pathophysiology of neuropsychiatric disorders.</p>

<p>The CNS monoaminergic systems are considered as the head engine regulating neuropsychiatric functions and personality. Transcription factor AP-2 is known to be essential for the development of the brainstem including the monoaminergic nuclei, and has the ability to regulate many genes in the monoaminergic systems. The ability of transcription factors to regulate specific gene expression, has lately made them hot candidates as drug targets. In this thesis, results indicating a role of AP-2 in the molecular effects of the antidepressant drugs citalopram and phenelzine, are presented. </p><p>A polymorphism in the second intron of the gene encoding AP-2ß has previously been associated with anxiety-related personality traits as estimated by the Karolinska Scales of Personality (KSP). In this thesis, results confirming this association, gained by using a larger material and several different personality scales, are presented. Furthermore, data is presented showing an association between the activity of platelet monoamine oxidase, a trait-dependent marker for personality, and the genotype of the AP-2ß intron 2 polymorphism. </p><p>The functional importance of the AP-2ß intron 2 polymorphism has not yet been elucidated. Included in this thesis are results showing that the AP-2ß intron 2 polymorphism is not in linkage disequilibrium with the only other described polymorphism in the AP-2ß gene, i.e. in the AP-2ß promoter (-67 G/A). Introns have in several studies been shown to include binding sites for regulatory proteins, and thus, to be important in transcriptional regulation. Results are presented demonstrating that one human brain nuclear protein binds only to the long variant of the AP-2ß intron 2 polymorphism. If this protein is involved in the regulation of the AP-2ß gene, it would affect the expression levels of the AP-2ß protein. </p><p>In general, this thesis further establishes the role of transcription factor AP-2 as a regulatory factor of importance for personality and monoaminergic functions.</p>

Eukaryotic gene transcription plays a regulatory role in mammalian developmental processes. It has been shown that transcriptional control is an important mechanism for specification of neurotransmitter phenotypes. In the mammalian central nervous system, the transcription factor AP-2 family is one of the critical regulatory factors for neural gene expression and neuronal development. It has been shown that several genes in the monoaminergic systems have AP-2 binding sites in regulatory regions, suggesting a regulatory role of AP-2 also in the adult brain. Brainstem monoamines are implicated in the expression of personality traits and imbalances in these systems may give rise to psychiatric disorders. The gene encoding AP-2β includes a polymorphic region consisting of a tetranucleotide repeat of [CAAA]4-5 in intron 2. Studies on AP-2β genotype in relation to personality and platelet MAO activity, a trait-dependant marker for personality, are presented in this thesis. Furthermore, correlations between brainstem levels of AP-2α and AP-2β and monoamine turnover in projection areas in rat forebrain are reported. These results strengthen the notion that the AP-2 family is important regulators of the monoaminergic systems in the adult brain. Furthermore, two studies are presented in this thesis with analyses indicating a role for AP-2 in the molecular mechanism of antidepressant drugs. Altogether, this thesis presents data supporting our notion that the transcription factor AP-2 family is involved in the regulation of the monoaminergic systems both pre- and postnatally, and, therefore, might be involved in the pathophysiology of neuropsychiatric disorders.

The CNS monoaminergic systems are considered as the head engine regulating neuropsychiatric functions and personality. Transcription factor AP-2 is known to be essential for the development of the brainstem including the monoaminergic nuclei, and has the ability to regulate many genes in the monoaminergic systems. The ability of transcription factors to regulate specific gene expression, has lately made them hot candidates as drug targets. In this thesis, results indicating a role of AP-2 in the molecular effects of the antidepressant drugs citalopram and phenelzine, are presented. A polymorphism in the second intron of the gene encoding AP-2ß has previously been associated with anxiety-related personality traits as estimated by the Karolinska Scales of Personality (KSP). In this thesis, results confirming this association, gained by using a larger material and several different personality scales, are presented. Furthermore, data is presented showing an association between the activity of platelet monoamine oxidase, a trait-dependent marker for personality, and the genotype of the AP-2ß intron 2 polymorphism. The functional importance of the AP-2ß intron 2 polymorphism has not yet been elucidated. Included in this thesis are results showing that the AP-2ß intron 2 polymorphism is not in linkage disequilibrium with the only other described polymorphism in the AP-2ß gene, i.e. in the AP-2ß promoter (-67 G/A). Introns have in several studies been shown to include binding sites for regulatory proteins, and thus, to be important in transcriptional regulation. Results are presented demonstrating that one human brain nuclear protein binds only to the long variant of the AP-2ß intron 2 polymorphism. If this protein is involved in the regulation of the AP-2ß gene, it would affect the expression levels of the AP-2ß protein. In general, this thesis further establishes the role of transcription factor AP-2 as a regulatory factor of importance for personality and monoaminergic functions.