Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Veronika Váchalová Supervisor: Prof. PharmDr. František Štaud, Ph.D. Consultant: Mgr. Rona Karahoda Title of diploma thesis: The influence of antidepressants on serotonin homeostasis in placenta Depression is a serious mental disorder affecting 10-20% of women during pregnancy. Up to 10% of these pregnant women are prescribed antidepressants (ADs), most frequently from the class of selective serotonin (5-HT) reuptake inhibitors (SSRIs). While the safety of this treatment is questionable due to reported impaired pregnancy/fetal outcomes, understanding of potential mechanistic causes is still lacking. During pregnancy, 5-HT is important for normal placental function and proper fetal development and programming. 5-HT homeostasis in the placenta is maintained via the 5-HT transporter (SERT/SLC6A4) on the apical side and the recently characterized organic cation transporter 3 (OCT3/SLC22A3) on the basal side of trophoblast. These transporters take up 5-HT from the maternal and fetal circulations, respectively into the syncytiotrophoblast (STB) where it is degraded by monoamine oxidase-A (MAO-A). As all ADs interfere with the 5-HT system it is important to study their potential interactions in the...

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Whilst antidepressants are widely prescribed, there is a large degree of variation between patients in terms of treatment outcomes. Furthermore, the mechanisms by which these drugs exert their effects remain unclear. In this thesis, genetic biomarkers of antidepressant outcomes have been explored, in order to better understand the molecular mechanisms underpinning effective antidepressant treatment. The research presented here use data from the GENDEP project, which is a large pharmacogenetic study of depressed patients receiving antidepressant treatment. Firstly, the pharmacological underpinnings of antidepressant-associated side effects were used to categorise these side effects and conduct a candidate gene analysis. Whilst a significant association between variation within the HTR2C gene and serotonergic side effects was found, the observation was not replicated in a second sample. Secondly, the role of variability in drug metabolism rates in treatment outcomes was investigated. Examining genotypic information on the cytochrome P450 enzymes, no associations with treatment response, side effects or study discontinuation were observed. Furthermore, serum concentrations of antidepressant were unrelated to treatment response or overall burden of side effects, predicting only a minority of specific side effects. Thirdly, transcriptomic changes with drug administration were explored in relation to treatment efficacy. Two genes were identified where changes in expression levels were significantly associated with treatment response amongst patients taking nortriptyline. Furthermore, using a network-based approach, changes in gene expression across one module of coexpressed genes showed significant correlation with symptom improvement; this biological network generalised across different antidepressant medications. Finally, genomic and transcriptomic data were combined, in an examination of the genetic control of gene expression. This analysis then was used to gain an insight into the molecular processes that link genotype to phenotype. The evidence presented within this thesis, when considered in combination with existing literature, highlights that antidepressant efficacy is a complex trait, influenced by many genes of small effect. Nevertheless, by layering together different levels of information, we can begin to dissect the molecular mechanisms involved in antidepressant action.

Objective: To determine: 1) 12-month prevalence rates of major depressive disorder (MDD) by race, comparing African Americans and Hispanics with whites, while controlling for covariates; 2) if there are any differences in treatment (i.e., pharmacotherapy, provider, and CAM) for MDD among African Americans, Hispanics and whites diagnosed with major depressive disorder (MDD) in the past 12 months; and 3) if there are any racial differences in DSM-IV diagnosed alcohol and/or drug abuse among those with a diagnosis of 12-month MDD. Methods: This retrospective study utilized data from the National Comorbidity Survey-Replication, which was designed to collect information on the mental health status of a nationally representative sample (n = 9282) of U.S. adults. Respondents with an MDD diagnosis in the past 12 months were included. The dependent variables were: 12-month MDD, pharmacotherapy, provider (mental health specialist), CAM and alcohol and/or drug abuse. Pharmacotherapy was examined by assessing respondents’ reported antidepressant use and whether the medication used was an SSRI/SNRI. The primary independent variable was race. Additional covariates included: age, gender, income, education, marital and employment status. Logistic regression was used to address the study objectives. Results: African Americans were significantly less likely to be diagnosed with MDD during the preceding 12-month period, when compared to whites, while controlling for covariates (OR= 0.6, p = 0.0169). Other significant variables for prevalence were female gender (OR= 1.8, p <0.0001), divorced/separated (OR= 2.0, p <0.0001) and not in the labor force (e.g., homemaker/retired) (OR= 1.5, p = 0.0033). Although African Americans and Hispanics reported lower antidepressant use, the results were not significant. With respect to SSRIs/SNRIs, African Americans reported significantly lower use, when compared to whites (OR= 0.3, p = 0.0309). Hispanics in the study were less likely to see a mental health professional (OR = 0.2, p = 0.0002). CAM use was significantly lower among individuals with less than 12 years, 12 years and 13 to 15 years of education (OR = 0.3, p = 0.0110; OR = 0.3, p = 0.0035; OR = 0.7, p = 0.0368, respectively) when compared to respondents with 16 years or more of education. When examining alcohol and/or drug use in those with diagnosed MDD in the preceding 12 months, females were less likely to be abusers (OR = 0.4, p = 0.0204). Conclusion: Among respondents with a diagnosis of MDD, race plays an important role in the types of treatment utilized to manage the disorder. Considering, the disability associated with depression, greater efforts are needed to improve antidepressant therapy for African Americans and mental health specialty treatment for Hispanics. / text

The purpose of this research project was to look for a neuropharmacological correlate to the behavioral deficits seen in learned helplessness (LH). The fact that antidepressant drugs reverse the deficits seen in a helpless rat, strongly suggests that the deficit is due to some neurochemical imbalance. This imbalance could be due to either the uncontrollable stressor or genetically induced. The three experiments suggest that there are fundamental differences in the way the CNS of helpless-prone rats and helpless-resistant rats cope with unpredictable and inescapable footshock. The goal of Experiment I was to search for a correlation between LH and receptor changes in the frontal cortex. The results did not support the hypothesis. The use of a heterogeneous stock of rat may have masked any basic differences between helpless-prone and helpless-resistant rats with regard to the 5-HT₂ and β-adrenergic receptors in the frontal cortex. Based on previous studies and the results from Experiment I, one could argue that there exists a genetic component in LH. The results from Experiment II suggest a strong genetic component to LH, not unlike that found in certain forms of human depressive disorders. Accordingly, rats from eight different stocks were tested for susceptibility to LH training. Of the eight stocks tested, Kyoto and Charles River Holtzman rats were the most susceptible at 53% and 55%, respectively. Overall, the variability ranged from 0% to 50%. These results indicate that wide differences in susceptibility to LH training exist in rats from different stocks or suppliers. The results of Experiment II suggested that the Kyoto Wistar rat would be a reliable inbred strain in which to study LH. With regards to the original goal of this research, it was decided that an evaluation of different neuro-transmitter systems during the LH paradigm would yield a potential for success in finding a biochemical marker that would differentiate LH-prone from LH-resistant rats. The results of Experiment III suggest, at least in hippocampus, that the serotonin (5-HT) and norepinephrine (NE) systems are differentially affected in the LH-prone and LH-resistant rat. In particular 5-HT levels are not affected by stress alone, but are increased in LH-prone rats following a frustrating test session. Also, the NE metabolite MHPG, is not affected by stress, but does increase in the LH-prone rat following testing. Both of these results differentiate the LH-prone and LH-resistant rat. In conclusion, the three experiments suggest that there is a genetic component in LH and that the NE and 5-HT systems are differentially affected by uncontrollable and inescapable shock in LH-prone and LH-resistant rats.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / IntroduÃÃo: DepressÃo à uma condiÃÃo psicopatolÃgica crÃnica e recorrente, com evidentes prejuÃzos sÃcio-ocupacionais e de qualidade de vida, tanto na fase aguda quanto no longo prazo. A depressÃo à causada por um dÃficit funcional das monoaminas nas suas vias transmissoras (serotoninÃrgicas, noradrenÃrgica e dopaminÃrgicas) em certos locais do cÃrebro. Apesar de existirem diversas formas de tratamento clÃnico desse transtorno do humor como: psicoterapia, antidepressivos, ECT e TMS, a prescriÃÃo das drogas antidepressivas representa a principal forma de manejo das chamadas depressÃes endÃgenas. A modulaÃÃo funcional das vias de transmissÃo neuroquÃmica pode interferir no desempenho do processamento de funÃÃes cognitivas como a memÃria. Objetivo: Nosso estudo procurou investigar as possÃveis alteraÃÃes em diferentes tipos e fases da memÃria provocadas pelo uso crÃnico de diferentes manejos de antidepressivos; monoterapia e terapia combinada. MÃtodos: Utilizamos nessa pesquisa a administraÃÃo crÃnica de trÃs drogas antidepressivas e suas associaÃÃes (Paroxetina, Venlafaxina e Bupropiona) de largo uso na prÃtica clÃnica. Para avaliaÃÃo do desempenho mnemÃnico utilizamos modelos comportamentais para ratos com alto valor preditivo para memÃria (labirinto radial, esquiva passiva) e para atividade exploratÃria (campo aberto). Foram medidos os nÃveis de BDNF e a atividade enzimÃtica da AChE do hipocampo dos grupos de ratos usuÃrios de antidepressivos. Resultados: No Labirinto Radial â fase de aquisiÃÃo: os grupos Venlaf (11Â4,85), Venlaf+Parox (10,88Â5,30) e Venlaf + Buprop (11Â3,85) apresentaram nÃmero de tentativas de treinos significativamente maior comparado ao desempenho dos animais do grupo controle (5,11Â1,36), enquanto os grupos com Parox (4,14Â2,19), Buprop (7,1Â3,66) sozinha e Parox + Bup (4,75Â1,75) nÃo apresentaram alteraÃÃes do nÃmero de tentativas significantes. Labirinto Radial â MC: os grupos Venlaf (2,16Â1,47), Venlaf+Parox (2,85Â1,35) e Venla + Bup (2,62Â1,76) nÃo apresentaram alteraÃÃes significantes em comparaÃÃo ao grupo usuÃrio da soluÃÃo salina (2,25Â1,58), enquanto que os usuÃrios dos antidepressivos Parox (3,11Â1,69), Bup (4,9Â2,99) e Parox + Bup (4,12Â2,58) apresentaram um nÃmero significativamente (p<0,001) maior de erros em relaÃÃo ao grupo controle (2,25Â1,58). Labirinto Radial â ML: grupos usuÃrios de Venla (4,5Â1,64), Buprop (4Â2,3) e Venlaf+ Bup (2,87Â1,72) nÃo apresentaram alteraÃÃes significativamente diferente do grupo controle (3.11Â1,36), enquanto que o grupo usuÃrio de Parox (4,4Â1,50) apresentou um nÃmero significativamente maior de erros em relaÃÃo ao grupo controle (3.11Â1,36). Os grupos usuÃrios de Venlaf + Parox (2,66Â0,85) e Parox+Buprop (3,12Â2,16) apresentaram um nÃmero de erros significativamente menor quando comparados ao nÃmero de erros na Parox (4,4Â1,50). Esquiva Passiva â MC e ML:os grupos usuÃrios de Parox (218,4Â113,1), Bup (39,57Â23,19) e Parox+Buprop (300Â0,0) apresentaram um tempo de latÃncia significativamente (p<0,001) maior em relaÃÃo ao grupo controle (270Â37,30). ML: os grupos usuÃrios de Venlaf+Parox (218,1Â103,6) e Parox+ Bup (296,7Â8,69) reverteram o aumento do tempo de latÃncia do grupo de Parox (218,1Â103,6) e causou uma diminuiÃÃo significativa (p<0,001) do tempo de permanÃncia do rato no compartimento claro em relaÃÃo ao mesmo grupo. Campo Aberto â N0 cruzamentos: Parox (20,80Â3,85) apresentou um aumento significativo do nÃmero de cruzamentos em relaÃÃo ao grupo controle (14,90Â3,85), enquanto que o grupo usuÃrio de Parox+Buprop (12Â2,50) reverteu o aumento da atividade locomotora do grupo da Parox (20,80Â3,85). ConclusÃes: no teste de aquisiÃÃo houve prejuÃzo de desempenho nos grupos usuÃrios de antidepressivos com pouca seletividade para recaptaÃÃo de serotonina. O desempenho da MC e ML, em habilidades visuo-espaciais, obtivemos dÃficits nos grupos de antidepressivos que potencializam tanto a aÃÃo serotoninÃrgica, com a aÃÃo dopaminÃrgica. A potencializaÃÃo &#946;-adrenÃrgica induziu uma melhora do desempenho da formaÃÃo e consolidaÃÃo da MC e ML. / Introduction: Depression is a chronic and recurrent psychopathological condition, with obvious socio-occupational injury and quality of life in both the acute and long term. Depression is caused by afunctional deficit of monoamines in their signaling pathways (serotoninergic, dopaminergic, noradrenergic) in certain parts of the brain. Although there are various forms of clinical treatment of mood disorders such as psychotherapy, antidepressants, ECT and TMS, the prescription of antidepressant drugs is the main form of management of calls endogenous depressions. The functional modulation of neurochemical transmission paths can interfere with the processing performance of cognitive functions like memory. Objective: Our study sought to investigate the possible changes in different types and stages of memory caused by chronic use of antidepressants from different managements; monotherapy and combination therapy. Methods: We use this research to chronic administration of three antidepressant drugs and their associations (paroxetine, venlafaxine and bupropion) widely used in clinical practice. To evaluate the performance mnemonic to use behavioral models predictive of mice with eigen value memory (radial maze, passive avoidance) and exploratory activity (open field). They measured levels of BDNF and AChE in the hippocampus of rats in groups of antidepressant users. Results: Radial Maze - acquisition phase: the Venlaf group (11 Â4.85), associated with Venlaf Parox (10,88Â5,30) associated with Buprop and Venlaf (11  3.85) showed a number of training trials significantly higher compared to that of the control group (5.11 1.36), while groups with Parox (4, 14  2.19), Buprop (7.1  3.66) associated with Parox alone and Buprop (4.75  1.75) showed no change in the number of significant attempts. Radial Maze - MC: Venlaf groups (2.16 Â1.47) associated with Venlaf Parox (2.85  1.35) associated with Buprop and Venlaf (2.62  1.76) showed no significant changes compared to the user group of saline (2.25  1.58) while users of antidepressants Parox (3.11  1.69), Buprop (4.9 2.99) associated with Buprop and Parox (4.12  2.58) had significantly (p <0.001) greater errors in the control group (2.25  1.58). Radial Maze â ML: Venlaf users groups (4.5  1.64), Buprop (4  2.3) associated with Buprop and Venlaf (2.87  1.72) showed no significant than the control group (3.11  1.36),where as the group user Parox (4.4 1.50) howed a significantly greater errors in the control group (3.11  1 ,36) Venlaf users groups associated with Parox (2.66  0.85) associated with Buprop and Parox (3.12  2.16) had a number of errors significantly lower when compared to the number of errors in Paroxetine (4.4  1.50). Avoidance passive âMC and ML : Parox users groups(218.4  113.1) Buprop (39.57  23.19) and associated with Parox Buprop (300  0.0) showed a significant lag time higher in the control group (270  37.30). ML: Venlaf users groups associated with Parox (218.1  103.6) associated with Buprop and Parox (296.7  8.69) revert the increase in the latency time of the Parox (218,1Â103,6) and caused a significant decrease the time spent in the compartment of the mouse course over the same group. Open Field: Parox (20.80  3.85) increased significantly in the control (14.90  3.85) while the user group Buprop associated with Parox (12  2.50) reversed the increase in locomotor activity of the Parox group (20.80  3.85). Conclusions: the test of acquisition show loss of performance in groups o antidepressant users with little selectivity for serotonin reuptake. The MC and ML in visual-spatial skills, we found deficits in groups of antidepressants that potentiate both serotonin action with dopaminergic action. The potentiation of &#946;-adrenergic is associated with increased level of alertness and improves the performance of establishment and consolidation of MC and ML.

Untreated depression in pregnancy may result in adverse health outcomes to both the mother and her unborn child. Pharmacotherapy with antidepressants is the most common treatment option for depression; however, the decision to treat with medication becomes complicated by pregnancy. Risk benefit assessments are critical tools to guide the treatment decision. Factors that should be included in these analyses include the pharmacokinetics and pharmacodynamics of antidepressants in pregnancy and their maternal and fetal safety. The economic cost of untreated maternal depression is also important to keep in mind. When the pharmacokinetics of the antidepressants venlafaxine and bupropion were studied in pregnancy it was found that the apparent oral clearance rate of bupropion was increased in late pregnancy when compared to early pregnancy (p = 0.03). There was a trend for lower area under the curve for these medications when the third trimester was compared to the first trimester. When the metabolism of antidepressants was investigated using hair analysis it was found that there was increased metabolism in pregnancy when compared to the postpartum period for citalopram (p = 0.02) but not venlafaxine (p = 0.77). Follow up of depressive symptoms throughout pregnancy identified that depression scores were highest in the first trimester of pregnancy, which may be due to concurrent nausea and vomiting of pregnancy. A meta-analysis of paroxetine use in early pregnancy demonstrated that there was no increased risk for cardiac malformations; case-control studies had an odds ratio of 1.18 (CI95: 0.88 – 1.59) while a weighted average difference of 0.3% was found in case-control studies (CI95: -0.1 – 0.7%, p = 0.19) The direct medical costs incurred by the Ontario government due to discontinuation of antidepressant medications in pregnancy was estimated to exceed $20,000,000 CAD. The management of depression in pregnancy with pharmacotherapy is an important and complex issue. My study documents the advantages of conducting risk benefit assessments for vulnerable populations such as pregnant women with depression.

Untreated depression in pregnancy may result in adverse health outcomes to both the mother and her unborn child. Pharmacotherapy with antidepressants is the most common treatment option for depression; however, the decision to treat with medication becomes complicated by pregnancy. Risk benefit assessments are critical tools to guide the treatment decision. Factors that should be included in these analyses include the pharmacokinetics and pharmacodynamics of antidepressants in pregnancy and their maternal and fetal safety. The economic cost of untreated maternal depression is also important to keep in mind. When the pharmacokinetics of the antidepressants venlafaxine and bupropion were studied in pregnancy it was found that the apparent oral clearance rate of bupropion was increased in late pregnancy when compared to early pregnancy (p = 0.03). There was a trend for lower area under the curve for these medications when the third trimester was compared to the first trimester. When the metabolism of antidepressants was investigated using hair analysis it was found that there was increased metabolism in pregnancy when compared to the postpartum period for citalopram (p = 0.02) but not venlafaxine (p = 0.77). Follow up of depressive symptoms throughout pregnancy identified that depression scores were highest in the first trimester of pregnancy, which may be due to concurrent nausea and vomiting of pregnancy. A meta-analysis of paroxetine use in early pregnancy demonstrated that there was no increased risk for cardiac malformations; case-control studies had an odds ratio of 1.18 (CI95: 0.88 – 1.59) while a weighted average difference of 0.3% was found in case-control studies (CI95: -0.1 – 0.7%, p = 0.19) The direct medical costs incurred by the Ontario government due to discontinuation of antidepressant medications in pregnancy was estimated to exceed $20,000,000 CAD. The management of depression in pregnancy with pharmacotherapy is an important and complex issue. My study documents the advantages of conducting risk benefit assessments for vulnerable populations such as pregnant women with depression.