DOCSLIB.ORG

Identifying the Mechanisms of Antidepressant Drug Action in Mice Lacking Brain Serotonin

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Identifying the mechanisms of antidepressant drug action in mice lacking brain serotonin D I S S E R T A T I O N zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) eingereicht an der Lebenswissenschaftlichen Fakultät der Humboldt-Universität zu Berlin von Markus Petermann Präsidentin der Humboldt-Universität zu Berlin Prof. Dr.-Ing. Dr. Sabine Kunst Dekan der Lebenswissenschaftlichen Fakultät der Humboldt-Universität zu Berlin Prof. Dr. Bernhard Grimm Gutachter 1. Prof. Dr. Michael Bader 2. Prof. Dr. Rüdiger Krahe 3. Prof. Dr. Golo Kronenberg Tag der mündlichen Prüfung: 08. Januar 2021 https://doi.org/10.18452/23035 EIDESSTATTLICHE ERKLÄRUNG ZUR SELBSTSTÄNDIGKEIT 2 SUMMARY 3 ZUSAMMENFASSUNG 4 1 INTRODUCTION 5 1.1 MAJOR DEPRESSIVE DISORDER 5 DEFINITION AND HISTORICAL DEVELOPMENT 5 HYPOTHESIS OF DEPRESSION 8 1.2 MONOAMINE HYPOTHESIS OF DEPRESSION 9 THE MONOAMINE SEROTONIN AND ITS HISTORICAL RELEVANCE 9 SEROTONIN BIOSYNTHESIS AND METABOLISM 9 PERIPHERAL SEROTONIN 11 THE NEUROTRANSMITTER SEROTONIN 12 NEUROANATOMY OF THE SEROTONIN SYSTEM IN MOUSE AND HUMAN BRAIN 13 SEROTONIN-RECEPTORS 14 SEROTONIN TRANSPORTER 15 ADULT NEUROGENESIS 18 SEROTONIN AS THERAPEUTIC TARGET 20 SELECTIVE SEROTONIN REUPTAKE INHIBITORS (SSRI) 22 SELECTIVE SEROTONIN REUPTAKE ENHANCER (SSRE) 23 ELECTROCONVULSIVE THERAPY 24 1.3 ANIMAL MODELS AS A TOOL FOR RESEARCHING THE SEROTONIN SYSTEM 26 TPH2 - DEFICIENT MICE 26 SERT - DEFICIENT MICE 29 1.4 NEUROTROPHIN HYPOTHESIS OF DEPRESSION 30 THE NEUROTROPHINS AND BDNF 30 BDNF BIOSYNTHESIS AND PATHWAYS 30 ALTERED BDNF SIGNALING IN DISEASE – ANTIDEPRESSANT EFFECTS OF BDNF 32 1.5 NEURO-ENDOCRINE SYMPTOM OF DEPRESSION AND THE STRESS HYPOTHESIS 33 PHYSIOLOGICAL STRESS 33 DYSREGULATION OF THE HPA-AXIS AND THE STRESS RECEPTORS 34 1.6 BIOMARKERS FOR DEPRESSION 36 1.7 AIMS 38 COMPARISON OF THE PHYSIOLOGICAL EFFECTS MEDIATED BY SSRI AND SSRE TREATMENT 38 TEST ELECTROCONVULSIVE THERAPY (ECT) IN A MODEL WITH SEROTONIN DEPLETION 38 ELUCIDATING THE ROLE OF SEROTONIN ON THE HPA-AXIS 38 ALTERATIONS IN DAILY LIFE ACTIVITIES AS A BEHAVIOR BIOMARKER FOR DEPRESSION 39 2 METHODS 40 2.1 ANIMALS AND HOUSING CONDITIONS 40 TPH2-/- 40 SERT-/- 40 ACE2-/- 40 HOUSING CONDITIONS 40 2.2 BRDU AND DRUG TREATMENT 41 BRDU TREATMENT 41 SSRI, SSRE TREATMENT 41 2.3 EXPERIMENTAL DESIGNS 42 COMPARISON OF THE PHYSIOLOGICAL EFFECTS MEDIATED BY SSRI AND SSRE TREATMENT 42 TEST ELECTROCONVULSIVE THERAPY (ECT) IN A MODEL WITH SEROTONIN DEPLETION 42 ELUCIDATING THE ROLE OF SEROTONIN ON THE HPA-AXIS 43 ALTERATIONS IN DAILY LIFE ACTIVITIES AS A BEHAVIOR BIOMARKER FOR DEPRESSION 44 2.4 GENOTYPING 44 GDNA ISOLATION 44 PCR-AMPLIFICATION OF THE TEMPLATE DNA 45 GEL ELECTROPHORESIS FOR DNA SEPARATION 46 2.5 TISSUE PREPARATION 46 ANESTHESIA OF ANIMALS 46 ANIMAL PERFUSION 47 ORGAN SECTIONING AND CRYOPROTECTION 47 TRUNK BLOOD COLLECTION AND PLASMA ISOLATION 47 2.6 HISTOLOGY / IMMUNOFLUORESCENCE 48 DAB-LABELING OF BRDU-POSITIVE CELLS 48 IMMUNOFLUORESCENCE OF BRDU-POSITIVE CELLS FOR PHENOTYPIC ANALYSIS 49 QUANTIFICATION OF CELL PROLIFERATION AND SURVIVAL 50 TUNNEL CELL DEATH STAINING 50 2.7 ELISA 51 STANDARD CURVE CALCULATION FOR ENDOCRINE HPA-PARAMETERS 51 BDNF 51 CORT / GLUCOCORTICOIDS 51 ACTH 52 2.8 QPCR FOR MRNA EXPRESSION OF HPA-AXIS PARAMETERS 52 COMPLEMENTARY DNA (FIRST STRAND) SYNTHESIS 52 QUANTITATIVE REAL-TIME PCR (QRT-PCR / QPCR) 53 SPECTROMETRIC MEASUREMENT OF NUCLEIC ACID CONCENTRATION 54 2.9 BEHAVIORAL TESTING 55 ELEVATED O-MAZE 55 OPEN FIELD 55 FORCED SWIM TEST 56 NESTING BEHAVIOR 56 HOARDING 57 BURROWING 58 2.10 VAGINAL SMEAR ANALYSIS TO DETERMINE ESTRUS CYCLE STAGES 58 2.11 STATISTICAL ANALYSIS 59 2.12 MISCELLANEOUS SOFTWARE 60 3 RESULTS 61 3.1 COMPARISON OF THE PHYSIOLOGICAL EFFECTS MEDIATED BY SSRI VS. SSRE TREATMENT 61 THE EFFECTS OF THE SSRI CIT ON ADULT NEUROGENESIS IN TPH2-/- MICE AND CONTROL 61 CELL SURVIVAL W/O TREATMENT 62 THE EFFECTS OF THE SSRE TIA ON ADULT NEUROGENESIS IN TPH2-/- MICE AND CONTROL 63 PHENOTYPIC ANALYSIS AND APOPTOSIS 65 3.1.5 CIT TREATMENT DECREASES BDNF PROTEIN LEVELS IN TPH2-/- MICE 69 3.2 DYSREGULATION OF THE HPA-AXIS IN THE ABSENCE OF SEROTONIN 71 CORT AND ACTH PLASMA LEVELS 71 CORT AND ACTH LEVELS UPON TREATMENT PARADIGMS 75 CRF AND POMC LEVELS IN HYPOTHALAMUS 77 NO INFLUENCE OF ESTROUS CYCLE ON CORT AND ACTH LEVELS 80 OPEN FIELD TEST UPON TREATMENT 81 GLUCOCORTICOID-, AND MINERALOCORTICOID RECEPTOR GENE EXPRESSION 83 3.3 ELECTROCONVULSIVE SEIZURE (ECS) 92 ECS DOES NOT AFFECT ANXIETY LEVELS BUT ATTENUATES THE HIGHLY ACTIVE PHENOTYPE OF TPH2-/- MICE 92 REDUCED NEUROBIOLOGICAL RESPONSES TO ECS IN TPH2-/- MICE 95 3.4 BEHAVIORAL SETUPS AS A BIOMARKER FOR DEPRESSION 97 REDUCED NESTING BEHAVIOR IN THE LACK OF SEROTONIN 97 HOARDING AND BURROWING 99 4 DISCUSSION 101 4.1 MONOAMINE HYPOTHESIS – SEROTONIN IS TARGET IN SSRI BUT NOT SSRE FUNCTION 101 STRESS EFFECTS ON ADULT NEUROGENESIS 104 THE SSRE TIA – A DOUBTFUL “HYPOTHESIS-KILLER” ! 106 4.2 NEUROTROPHIN HYPOTHESIS – BDNF IS INVOLVED IN BRAIN PLASTICITY IN THE ABSENCE OF SEROTONIN 108 4.3 SEROTONIN AFFECTS THE HPA-AXIS 111 RECEPTORS OF THE STRESS AXIS 114 SEX-SPECIFIC PHYSIOLOGICAL FUNCTIONS OF SEROTONIN 117 SERT−/− MICE ARE PERFECT COUNTERPARTS FOR THE STRESS-RESULT FROM TPH2-/- MICE 119 4.4 ELECTROCONVULSIVE THERAPY (ECT) IN A MODEL WITH SEROTONIN DEPLETION 121 4.5 BEHAVIOR AS BIOMARKER 124 4.6 CONCLUSION 126 5 MATERIAL 128 5.1 ANTIBODIES 130 5.2 OLIGONUCLEOTIDES (PRIMERS) 130 5.3 LABORATORY EQUIPMENT 132 5.4 SOFTWARE 133 6 ABBREVIATIONS 134 7 BIBLIOGRAPHY 137 8 LIST OF TABLES 160 9 TABLE OF FIGURES 162 Dedicated to Sven, Rike & Theresa 1 Eidesstattliche Erklärung zur Selbstständigkeit Eidesstattliche Erklärung zur Selbstständigkeit Hiermit erkläre ich, dass ich die vorliegende Arbeit selbstständig und nur unter der Verwendung der angegebenen Quellen und Hilfsmittel angefertigt habe. Alle Materialien oder Dienstleistungen, die ich von Dritten erhalten habe, sind als solche gekennzeichnet. Ich versichere, dass ich mich nicht anderswertig um einen Doktorgrad beworben habe oder einen entsprechenden Doktortitel bereits besitze. Die Promotionsordnung der Lebenswissenschaftlichen Fakultät (Stand 5. März 2015) der Humboldt-Universität zu Berlin ist mir bekannt. __________________________ Markus Petermann Berlin; 01. September 2020 2 Summary Summary Serotonin is well-known as the "molecule of happiness" and the most important target for antidepressants. The mainly prescribed drugs in major depression are selective serotonin re-uptake inhibitors (SSRI); but recently, SSR-enhancer (SSRE) have also attracted clinical attention. However, only a quarter of patients responds to treatment and research into the drug mechanisms revives. It needs to be determined, whether SSRI/E act solely via manipulating serotonin levels or whether other pathways are involved, e.g. neurotrophic signaling (brain-derived neurotrophic factor, BDNF) or the hypothalamus-pituitary-adrenal (HPA)-axis. Furthermore, in major depression, dysregulation of central serotonin signaling is accompanied with a decline in hippocampal neurogenesis, as has been observed in rodent models. Therefore, to elucidate the interplay of serotonin, BDNF, adult neurogenesis and the HPA-axis is of importance. At the center of this thesis is a mouse model deficient in the central serotonin- synthesizing enzyme, tryptophan hydroxylase 2 (Tph2-/- mice). Using molecular biology tools, treatment, and behavioral paradigms, I have investigated physiological responses to antidepressant treatment in the absence of brain serotonin, and the possible role of alternative pathways. I observed the typical increase in neurogenesis upon SSRI treatment in WT mice, while it had no effect in Tph2-/- mice. In contrast, BDNF levels were significantly decreased in Tph2-/- mice after treatment with no effect in WT control mice. Furthermore, my results show a critical role of brain serotonin in the neurobiological effects of electroconvulsive seizure. Surprisingly, in animals lacking central serotonin, increased neurogenesis was observed independently of the treatment. The gathered data indicated an altered stress response; therefore, parameters of the HPA-Axis have been studied, indicating a downregulated HPA system in Tph2-/- animals in baseline state, but showed no difference in treatment or feedback control. This thesis gives insight into the mechanisms of antidepressant action and reveals ideas for novel pathways involved in the process that could be used as targets in therapeutic approaches and further research in major depression. 3 Zusammenfassung Zusammenfassung Serotonin ist bekannt als das "Glückshormon" und gilt als Hauptangriffsstelle der heute gängigsten Antidepressiva. Die am häufigsten verschriebenen Medikamente bei schweren Depressionen sind selektive Serotonin-Wiederaufnahmehemmer (SSRI). Dennoch erzeugten aufgrund guter Verträglichkeit im letzten Jahrzehnt aber auch SSR-Enhancer (SSRE) klinische Aufmerksamkeit. Nur etwa ein Viertel der Patienten sprechen auf therapeutische antidepressive Behandlungen an, weswegen eine große Notwendigkeit des Verständnisses ihrer Wirkungsweisen besteht. Es bleibt offen, ob SSRI / E ausschließlich über die Manipulation des Serotoninspiegels wirken, oder ob alternative Signalwege daran beteiligt sind. Ansatzpunkte hierfür sind beispielsweise die neurotrophen Signalwege (besonders Brain derived neurotophic factor, BDNF) oder die Hypothalamus- Hypophysen-Nebennieren- (HPA) – Signalwege des Stressachsensystems. Ebenfalls wurde in Nagetiermodellen beobachtet, dass mit der Dysregulation des zentralen
Recommended publications
  • Deamidation of Human Proteins

    Deamidation of Human Proteins

    Deamidation of human proteins N. E. Robinson*† and A. B. Robinson‡ *Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and ‡Oregon Institute of Science and Medicine, Cave Junction, OR 97523 Communicated by Frederick Seitz, The Rockefeller University, New York, NY, August 31, 2001 (received for review May 8, 2001) Deamidation of asparaginyl and glutaminyl residues causes time- 3D structure is known (23). This method is more than 95% dependent changes in charge and conformation of peptides and reliable in predicting relative deamidation rates of Asn residues proteins. Quantitative and experimentally verified predictive cal- within a single protein and is also useful for the prediction of culations of the deamidation rates of 1,371 asparaginyl residues in absolute deamidation rates. a representative collection of 126 human proteins have been It is, therefore, now possible to compute the expected deami- performed. These rates suggest that deamidation is a biologically dation rate of any protein for which the primary and 3D relevant phenomenon in a remarkably large percentage of human structures are known, except for very long-lived proteins. These proteins. proteins require measurement of the 400 Gln pentapeptide rates. in vivo deamidation ͉ asparaginyl residues Materials and Methods Calculation Method. The Brookhaven Protein Data Bank (PDB) eamidation of asparaginyl (Asn) and glutaminyl (Gln) was searched to select 126 human proteins of general biochem- Dresidues to produce aspartyl (Asp) and glutamyl (Glu) ical interest and of known 3D structure without bias toward any residues causes structurally and biologically important alter- known data about their deamidation, except for 13 proteins (as ations in peptide and protein structures.
  • Neuroenhancement in Healthy Adults, Part I: Pharmaceutical

    Neuroenhancement in Healthy Adults, Part I: Pharmaceutical

    l Rese ca arc ni h li & C f B o i o l e Journal of a t h n Fond et al., J Clinic Res Bioeth 2015, 6:2 r i c u s o J DOI: 10.4172/2155-9627.1000213 ISSN: 2155-9627 Clinical Research & Bioethics Review Article Open Access Neuroenhancement in Healthy Adults, Part I: Pharmaceutical Cognitive Enhancement: A Systematic Review Fond G1,2*, Micoulaud-Franchi JA3, Macgregor A2, Richieri R3,4, Miot S5,6, Lopez R2, Abbar M7, Lancon C3 and Repantis D8 1Université Paris Est-Créteil, Psychiatry and Addiction Pole University Hospitals Henri Mondor, Inserm U955, Eq 15 Psychiatric Genetics, DHU Pe-psy, FondaMental Foundation, Scientific Cooperation Foundation Mental Health, National Network of Schizophrenia Expert Centers, F-94000, France 2Inserm 1061, University Psychiatry Service, University of Montpellier 1, CHU Montpellier F-34000, France 3POLE Academic Psychiatry, CHU Sainte-Marguerite, F-13274 Marseille, Cedex 09, France 4 Public Health Laboratory, Faculty of Medicine, EA 3279, F-13385 Marseille, Cedex 05, France 5Inserm U1061, Idiopathic Hypersomnia Narcolepsy National Reference Centre, Unit of sleep disorders, University of Montpellier 1, CHU Montpellier F-34000, Paris, France 6Inserm U952, CNRS UMR 7224, Pierre and Marie Curie University, F-75000, Paris, France 7CHU Carémeau, University of Nîmes, Nîmes, F-31000, France 8Department of Psychiatry, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Eschenallee 3, 14050 Berlin, Germany *Corresponding author: Dr. Guillaume Fond, Pole de Psychiatrie, Hôpital A. Chenevier, 40 rue de Mesly, Créteil F-94010, France, Tel: (33)178682372; Fax: (33)178682381; E-mail: [email protected] Received date: January 06, 2015, Accepted date: February 23, 2015, Published date: February 28, 2015 Copyright: © 2015 Fond G, et al.
  • Traxoprodil, a Selective Antagonist of the NR2B Subunit of the NMDA

    Traxoprodil, a Selective Antagonist of the NR2B Subunit of the NMDA

    Metab Brain Dis (2016) 31:803–814 DOI 10.1007/s11011-016-9810-5 ORIGINAL ARTICLE Traxoprodil, a selective antagonist of the NR2B subunit of the NMDA receptor, potentiates the antidepressant-like effects of certain antidepressant drugs in the forced swim test in mice Ewa Poleszak1 & Weronika Stasiuk 2 & Aleksandra Szopa1 & Elżbieta Wyska3 & Anna Serefko1 & Anna Oniszczuk4 & Sylwia Wośko 1 & Katarzyna Świąder1 & Piotr Wlaź5 Received: 25 November 2015 /Accepted: 17 February 2016 /Published online: 29 February 2016 # The Author(s) 2016. This article is published with open access at Springerlink.com Abstract One of the newest substances, whose antidepres- these changes were not due to the severity of locomotor ac- sant activity was shown is traxoprodil, which is a selective tivity. The observed effect of traxoprodil is only partially as- antagonist of the NR2B subunit of the NMDA receptor. The sociated with serotonergic system and is independent of the main goal of the present study was to evaluate the effect of effect on the 5-HT1A and 5-HT2 serotonin receptors. The re- traxoprodil on animals’ behavior using the forced swim test sults of an attempt to assess the nature of the interaction be- (FST), as well as the effect of traxoprodil (10 mg/kg) on the tween traxoprodil and the tested drugs show that in the case of activity of antidepressants, such as imipramine (15 mg/kg), joint administration of traxoprodil and fluoxetine, imip- fluoxetine (5 mg/kg), escitalopram (2 mg/kg) and reboxetine ramine or escitalopram, there were interactions in the (2.5 mg/kg). Serotonergic lesion and experiment using the pharmacokinetic phase.
  • Gbeta Paperv6

    Gbeta Paperv6

    bioRxiv preprint doi: https://doi.org/10.1101/415935; this version posted March 13, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 An interaction between Gβγ and RNA polymerase II regulates transcription in cardiac 2 fibroblasts 3 4 Shahriar M. Khan†, Ryan D. Martin†, Sarah Gora, Celia Bouazza, Jace Jones-Tabah, Andy 5 Zhang, Sarah MacKinnon, Phan Trieu, Paul B.S. Clarke, Jason C. Tanny*, and Terence E. 6 Hébert* 7 8 1Department of Pharmacology and Therapeutics, McGill University, Montréal, Québec, H3G 9 1Y6, Canada 10 11 12 †These authors contributed equally to the study. 13 14 15 *To whom correspondence should be addressed. 16 Dr. Terence E. Hébert, PhD, 17 Department of Pharmacology and Therapeutics, 18 McGill University, 19 3655 Promenade Sir-William-Osler, Room 1303 20 Montréal, Québec, H3G 1Y6, Canada 21 Tel: (514) 398-1398 22 E-mail: [email protected] 23 24 OR 25 26 Dr. Jason C. Tanny, PhD, 27 Department of Pharmacology and Therapeutics, 28 McGill University, 29 3655 Promenade Sir-William-Osler, Room 1303 30 Montréal, Québec, H3G 1Y6, Canada 31 Tel: (514) 398-3608 32 E-mail: [email protected] 33 1 bioRxiv preprint doi: https://doi.org/10.1101/415935; this version posted March 13, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 34 SUMMARY 35 Gβγ subunits are involved in many different signalling processes in various 36 compartments of the cell, including the nucleus.
  • Protein Symbol Protein Name Rank Metric Score 4F2 4F2 Cell-Surface

    Protein Symbol Protein Name Rank Metric Score 4F2 4F2 Cell-Surface

    Supplementary Table 2 Supplementary Table 2. Ranked list of proteins present in anti-Sema4D treated macrophage conditioned media obtained in the GSEA analysis of the proteomic data. Proteins are listed according to their rank metric score, which is the score used to position the gene in the ranked list of genes of the GSEA. Values are obtained from comparing Sema4D treated RAW conditioned media versus REST, which includes untreated, IgG treated and anti-Sema4D added RAW conditioned media. GSEA analysis was performed under standard conditions in November 2015. Protein Rank metric symbol Protein name score 4F2 4F2 cell-surface antigen heavy chain 2.5000 PLOD3 Procollagen-lysine,2-oxoglutarate 5-dioxygenase 3 1.4815 ELOB Transcription elongation factor B polypeptide 2 1.4350 ARPC5 Actin-related protein 2/3 complex subunit 5 1.2603 OSTF1 teoclast-stimulating factor 1 1.2500 RL5 60S ribomal protein L5 1.2135 SYK Lysine--tRNA ligase 1.2135 RL10A 60S ribomal protein L10a 1.2135 TXNL1 Thioredoxin-like protein 1 1.1716 LIS1 Platelet-activating factor acetylhydrolase IB subunit alpha 1.1067 A4 Amyloid beta A4 protein 1.0911 H2B1M Histone H2B type 1-M 1.0514 UB2V2 Ubiquitin-conjugating enzyme E2 variant 2 1.0381 PDCD5 Programmed cell death protein 5 1.0373 UCHL3 Ubiquitin carboxyl-terminal hydrolase isozyme L3 1.0061 PLEC Plectin 1.0061 ITPA Inine triphphate pyrophphatase 0.9524 IF5A1 Eukaryotic translation initiation factor 5A-1 0.9314 ARP2 Actin-related protein 2 0.8618 HNRPL Heterogeneous nuclear ribonucleoprotein L 0.8576 DNJA3 DnaJ homolog subfamily
  • Potential Roles of NCAM/PSA-NCAM Proteins in Depression and The

    Potential Roles of NCAM/PSA-NCAM Proteins in Depression and The

    Pharmacological Reports Copyright © 2013 2013, 65, 14711478 by Institute of Pharmacology ISSN 1734-1140 Polish Academy of Sciences Review PotentialrolesofNCAM/PSA-NCAMproteins indepressionandthemechanismofaction ofantidepressantdrugs KrzysztofWêdzony,AgnieszkaChocyk,MarzenaMaækowiak Laboratory of Pharmacology and Brain Biostructure, Department of Pharmacologcy, Institute of Pharmacology, Polish Academy of Sciences, Smêtna 12, PL 31-343 Kraków, Poland Correspondence: Krzysztof Wêdzony, e-mail: [email protected] Abstract: Recently, it has been proposed that abnormalities in neuronal structural plasticity may underlie the pathogenesis of major depression, resulting in changes in the volume of specific brain regions, including the hippocampus (HIP), the prefrontal cortex (PC), and the amygdala (AMY), as well as the morphology of individual neurons in these brain regions. In the present survey, we compile the data regarding the involvement of the neural cell adhesion molecule (NCAM) protein and its polysialylated form (PSA-NCAM) in the pathogenesis of depression and the mechanism of action of antidepressant drugs (ADDs). Elevated expression of PSA-NCAM may reflect neuroplastic changes, whereas decreased expression implies a rigidification of neuronal morphology and an impedance of dy- namic changes in synaptic structure. Special emphasis is placed on the clinical data, genetic models, and the effects of ADDs on NCAM/PSA-NCAM expression in the brain regions in which these proteins are constitutively expressed and neurogenesis is not a major factor; this emphasis is necessary to prevent cell proliferation and neurogenesis from obscuring the issue of brain plasticity. Keywords: antidepressantdrugs,depression,NCAM,PSA-NCAM Abbreviations: ADD – antidepressant drug, ADDs – antide- ogical effect of these drugs, i.e., the blockade of sero- pressants, antidepressant drugs, AMY – amygdala, FGFR – fi- tonin and noradrenaline uptake, is not clearly associ- broblast growth factor receptor, FLU – fluoxetine, HIP – hip- ated with their clinical efficacy [30].
  • Recombinant Rnasin(R)

    Recombinant Rnasin(R)

    Certificate of Analysis Recombinant RNasin® Ribonuclease Inhibitor: Part No. Size (units) Part# 9PIN251 N2511 2,500 Revised 4/18 N2515 10,000 Enzyme Storage Buffer: Recombinant RNasin® Ribonuclease Inhibitor is supplied in 20mM HEPES-KOH (pH 7.6), 50mM KCl, 8mM DTT, 50% (v/v) glycerol. Storage Conditions: See the Product Information Label for storage recommendations. Avoid multiple freeze-thaw cycles and exposure to frequent temperature changes. See the expiration date on the Product Information Label. Source: E. coli cells expressing a recombinant clone. *AF9PIN251 0418N251* Unit Definition: One unit is defined as the amount of Recombinant RNasin® Ribonuclease Inhibitor required to inhibit the AF9PIN251 0418N251 activity of 5ng of ribonuclease A by 50%. Activity is measured by the inhibition of hydrolysis of cytidine 2´,3´-cyclic monophosphate by ribonuclease A. The unit concentration is listed on the Product Information Label. Usage Notes: Recombinant RNasin® Ribonuclease Inhibitor is active over a broad pH range. Concentration gradients may form in frozen products and should be dispersed upon thawing. Mix well prior to use. Table 1. Properties of Recombinant RNasin® Ribonuclease Inhibitor. Property Comment Activity Inactivates RNase by noncovalent binding Molecular weight 49,847 daltons Type of inhibition Noncompetitive (3) Isoelectric point pI 4.7 pH activity range pH 5.5–9 (4) Binding ratio with RNase A 1:1 (3) Promega Corporation –14 Constant for binding inhibition Ki = 4 × 10 M (3,4) 2800 Woods Hollow Road Amount to use 1 unit of inhibitor per microliter of solution Madison, WI 53711-5399 USA Reaction conditions to avoid Temperatures >50°C, urea, SDS, other denaturants Telephone 608-274-4330 Toll Free 800-356-9526 Table 2.
  • Mechanism of Actions of Antidepressants: Beyond the Receptors

    Mechanism of Actions of Antidepressants: Beyond the Receptors

    Derlemeler/Reviews Mechanism of Actions of Antidepressants: Beyond the Receptors Mechanism of Actions of Antidepressants: Beyond the Receptors Ayflegül Y›ld›z, M.D.1, Ali Saffet Gönül, M.D.2, Lut Tamam, M.D.3 ABSTRACT: MECHANISM OF ACTIONS OF ANTIDEPRESSANTS: BEYOND THE RECEPTORS Since the discovery of first antidepressants-monoamine oxidase inhibitors- a half century passed. There are now almost two- dozen antidepressant agents that work by nine distinct pharmacological mechanisms at the receptor level. However, opposite to the divergence in their pharmacological mechanisms at the receptor level, antidepressant drugs probably stimulate similar pathways in subcellular level. These subcellular events or so called beyond receptor effects are named neuroplasticity, and the mechanism may be called as adaptation. These after-receptor processes, through their effects on synaptic transmission, and gene expression are indeed capable of altering many molecular events in the brain. In this article, the mechanisms of actions of antidepressants at- and beyond- the receptors are discussed by documenting some of the evidence indicating such long-term alterations. Accordingly, the well-known effects of antidepressants on the receptor level are initiating events of antidepressant drug action, which enhance and prolong the actions of norepinephrine and/or serotonin and/or dopamine. Only if an adequate dose of an antidepressant is taken chronically, the increase in the synaptic norepinephrine and/or serotonin and/or dopamine stresses or perturbs the nervous
  • Non-Conventional Effects of Antidepressants in the Central and Peripheral Nervous System

    Non-Conventional Effects of Antidepressants in the Central and Peripheral Nervous System

    Non-conventional effects of antidepressants in the central and peripheral nervous system Doctoral Thesis Aliz Mayer, M.D. Semmelweis University Szentágothai János Neuroscience Doctoral School Supervisor: Dr. János Kiss, Ph.D. Opponents: Dr. László Köles, Ph.D. Dr. Attila Kőfalvi, Ph.D. Exam Committee, President: Prof. Valéria Kecskeméti, full professor, Ph.D. Committee members: Dr. László Tretter, associate professor, Ph.D. Dr. László Hársing, scientific councillor, doctor of HAS. Budapest 2009 INTRODUCTION At the present time, depression is one of the most common neuropsychiatric disorders. According to the study of World Health Organization published in 2007, the 3.2 % of investigated population suffered from depression alone, but an average between 9.3% and 23.0% of participants had one or more chronic physical disease beside the depression. During a multicentric european study it was found, that the prevalence of depression in european adult population in 2001 it was approximately 8.56%. In all countries the rate of depressed women’s patients was higher than that of the number of depressed men’s patients. The epidemiology data of depression in Hungary shows similar rates: in 1998 the lifetime rate for major depressive disorder was 15.1%, and for bipolar disorders 5.1%. The female-to-male ratio was 2.7 for major depression. Nevertheless, today more than 30 antidepressants, with favourable side-effects profile are in the clinical use. The rate of efficient treatments don’t beyonds more than 50-60%, which means, that approximately 40% of the patients is not responder to the initial medication. The clinical efficacy of the latest antidepressants it isn’t better than that of the substances used 40 years ago, only one significant difference could be discovered between them: the favorable side-efects profile.
  • (12) United States Patent (10) Patent No.: US 9,504,665 B2 Cleveland (45) Date of Patent: *Nov

    (12) United States Patent (10) Patent No.: US 9,504,665 B2 Cleveland (45) Date of Patent: *Nov

    USOO9504665B2 (12) United States Patent (10) Patent No.: US 9,504,665 B2 Cleveland (45) Date of Patent: *Nov. 29, 2016 (54) HIGH-DOSE GLYCINE AS A TREATMENT USPC .................................................. 514/576,561 FOR OBSESSIVE-COMPULSIVE DSORDER See application file for complete search history. AND OBSESSIVE-COMPULSIVE SPECTRUM DSORDERS (56) References Cited (71) Applicant: W. Louis Cleveland, New York, NY U.S. PATENT DOCUMENTS (US) 6,030,604. A 2/2000 Trofast et al. 6,900, 173 B2 5/2005 Martin et al. (72) Inventor: W. Louis Cleveland, New York, NY 8,629, 105 B2 1/2014 Heresco-Levy et al. (US) 2004/O157926 A1 8/2004 Heresco-Levy et al. 2005, 0181019 A1 8, 2005 Palmer et al. (*) Notice: Subject to any disclaimer, the term of this 2006,0078593 A1 4, 2006 Strozier et al. patent is extended or adjusted under 35 U.S.C. 154(b) by 15 days. OTHER PUBLICATIONS This patent is Subject to a terminal dis- Wu, Master's Thesis, Graduate Institute of Medical Science, Chi claimer. nese Medical University, Jul. 2007.* English translation of Wu, Master's Thesis, Graduate Institute of (21) Appl. No.: 14/022,804 Medical Science, Chinese Medical University, Jul. 2007.* 9 Barco A. et al., “Common molecular mechanisms in explicit and 1-1. implicit memory.”, J Neurochem. 2006; vol. 97(6), pp. 1520-1533. (22) Filed: Sep. 10, 2013 Pittinger, C. et al., “In Search of general mechanisms for long O O lasting plasticity: Aplysia and the hippocampus’. Philos Trans R (65) Prior Publication Data Soc Lond B Biol Sci. 2003; vol.
  • Ribonuclease Inhibitors in Malus X Domestica (Common Apple): Isolation and Partial Characterization

    Ribonuclease Inhibitors in Malus X Domestica (Common Apple): Isolation and Partial Characterization

    Biosci. Biotechnol. Biochem., 67 (4), 698–703, 2003 Ribonuclease Inhibitors in Malus x domestica (Common Apple): Isolation and Partial Characterization Takao KOSUGE,1 Mamoru ISEMURA,2 Yoshiaki TAKAHASHI,3 Sumiko ODANI,4 and Shoji ODANI1,† 1Department of Biology, Faculty of Science, Niigata University, Ikarashi, Niigata 950-2181, Japan 2Department of Cellular Biochemistry, School of Food and Nutritional Sciences, University of Shizuoka, Tanida, Shizuoka 422-8526, Japan 3Department of Medical Technology, School of Health Science, Faculty of Medicine, Niigata University, Asahimachi, Niigata 951-8518, Japan 4Department of Home Economics, Faculty of Education and Human Science, Niigata University, Ikarashi, Niigata 950-2181, Japan Received August 22, 2002; Accepted January 6, 2003 A ribonuclease inhibitory activity was detected in the sion, and carbohydrate- and ATP-binding.1) Self- fruits of common apple, Malus x domestica,cv.Fuji, incompatibility in the ‰owering plants is also and puriˆed by a‹nity chromatography on ribonuclease mediated by prevention of self-pollination by S- A-Sepharose. It inhibited hydrolysis of cyclic-2?:3?- ribonuclease, which is a homologue of ribonuclease CMP by bovine pancreatic ribonuclease A with an T2.2) Therefore, a collective name of RISBASES apparent inhibition constant of about 5×10－8 M. (RIbonucleases with Special Biological Actions) was 1) Matrix-assisted laser desorptionWionization time-of- proposed for these RNase homologues. These ‰ight mass spectrometry of the puriˆed protein gave two diverse activities must be under precise control peaks corresponding to the mass numbers of 55,658 and responding to individual physiological conditions, 62,839, while three bands of 43-, 34-, and 21-kDa were but their mechanisms seem not fully understood.
  • Stress-Induced Hyperthermia, the Serotonin System and Anxiety

    Stress-Induced Hyperthermia, the Serotonin System and Anxiety

    The Open Pharmacology Journal, 2010, 4, 15-29 15 Open Access Stress-Induced Hyperthermia, the Serotonin System and Anxiety *,1 1,2 3 1 Christiaan H. Vinkers , Berend Olivier , J. Adriaan Bouwknecht , Lucianne Groenink and Jocelien D.A. Olivier4,5 1Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences (UIPS) and Rudolf Magnus Institute of Neuroscience, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht, The Netherlands 2Department of Psychiatry, Yale University School of Medicine, New Haven, USA 3Department of Neuroscience, Pharmaceutical Research & Development, Johnson and Johnson, Beerse, Belgium 4Department of Molecular Animal Physiology, Radboud University Nijmegen, Nijmegen, The Netherlands 5Donders Institute for Brain, Cognition and Behavior: Department for Neuroscience, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands Abstract: The serotonin (5-HT) system plays a key role in the pathophysiology of psychiatric disorders including mood and anxiety disorders. A role for serotonin in stress-related disorders is further supported by the fact that clinically effective treatments for these disorders alter serotonergic neurotransmission. The therapeutic potential of serotonergic pharmacological interventions has resulted in a variety of preclinical approaches to study the serotonin system. Of these, the stress-induced hyperthermia (SIH) paradigm has been extensively used to study the serotonin system at a preclinical level. The SIH response uses the transient rise in body temperature in response to a stressor which can be reduced using anxiolytic drugs including benzodiazepines, CRF receptor antagonists and serotonergic ligands. The present review aims to discuss the acute and chronic effects of 5-HT ligands on the SIH response. Also, the SIH response in genetically modified mice that lack or overexpress specific serotonergic receptor subtypes or the serotonin transporter will be summarized.