TAAR1 Activation Modulates Monoaminergic Neurotransmission, Preventing Hyperdopaminergic and Hypoglutamatergic Activity

Total Page:16

File Type:pdf, Size:1020Kb

TAAR1 Activation Modulates Monoaminergic Neurotransmission, Preventing Hyperdopaminergic and Hypoglutamatergic Activity TAAR1 activation modulates monoaminergic neurotransmission, preventing hyperdopaminergic and hypoglutamatergic activity Florent G. Revela, Jean-Luc Moreaua, Raul R. Gainetdinovb, Amyaouch Bradaiac, Tatyana D. Sotnikovab, Roland Morya, Sean Durkina, Katrin Groebke Zbindend, Roger Norcrossd, Claas A. Meyere, Veit Metzlera, Sylvie Chaboza, Laurence Ozmena, Gerhard Trubea, Bruno Pouzeta, Bernhard Bettlerf, Marc G. Carong, Joseph G. Wettsteina, and Marius C. Hoenera,1 aNeuroscience Research, dDiscovery Chemistry, and eDiscovery Technologies, Pharmaceuticals Division, F. Hoffmann-La Roche Ltd., CH-4070 Basel, Switzerland; bDepartment of Neuroscience and Brain Technologies, Italian Institute of Technology, 16163 Genoa, Italy; cNeuroservice, Domaine de Saint-Hilaire, 13593 Aix-en-Provence, France; fDepartment of Biomedicine, Institute of Physiology, Pharmazentrum, University of Basel, CH-4056 Basel, Switzerland; and gDepartment of Cell Biology, Duke University Medical Center, Durham, NC 27710 Edited by Richard D. Palmiter, University of Washington, Seattle, WA, and approved March 31, 2011 (received for review February 24, 2011) The trace amine-associated receptor 1 (TAAR1), activated by en- sitive to the locomotor-stimulating effect of d-amphetamine dogenous metabolites of amino acids like the trace amines and show elevated striatal release of dopamine (DA), noradren- p-tyramine and β-phenylethylamine, has proven to be an impor- aline (NA), and serotonin [5-hydroxytryptamine (5-HT)] after a d-amphetamine challenge (10, 12). Furthermore, the spontaneous tant modulator of the dopaminergic system and is considered −/− firing rate of the VTA DA neurons is augmented in Taar1 mice, a promising target for the treatment of neuropsychiatric disorders. fi To decipher the brain functions of TAAR1, a selective TAAR1 ago- and only in WT mice does pTyr decrease this ring rate (10). These fi observations suggest that TAAR1 is a negative modulator of mono- nist, RO5166017, was engineered. RO5166017 showed high af nity aminergic neurotransmission. and potent functional activity at mouse, rat, cynomolgus monkey, Apart from the TAs, TAAR1 is activated by a range of endoge- and human TAAR1 stably expressed in HEK293 cells as well as high nous molecules such as other biogenic amines (2, 5), thyroid selectivity vs. other targets. In mouse brain slices, RO5166017 PHARMACOLOGY hormone-derivative 3-iodothyronamine (T1AM) (13, 14), and inhibited the firing frequency of dopaminergic and serotonergic catechol-O-methyl transferase products (e.g., 3-methoxytyramine) neurons in regions where Taar1 is expressed (i.e., the ventral teg- (2, 15) or by synthetic substances such as amphetamine derivatives mental area and dorsal raphe nucleus, respectively). In contrast, and ergolines (2, 12). However, all these ligands have TAAR1- RO5166017 did not change the firing frequency of noradrenergic independent effects through other targets, such as the mono- neurons in the locus coeruleus, an area devoid of Taar1 expression. aminergic transporters and receptors or the σ-receptors (3, 11, 13, Furthermore, modulation of TAAR1 activity altered the desensiti- 16). The lack of selective ligands has rendered identification of zation rate and agonist potency at 5-HT receptors in the dorsal TAAR1 biological functions challenging. Recently, Bradaia et al. 1A (17) described the first selective TAAR1 antagonist, N-(3-Ethoxy- raphe, suggesting that TAAR1 modulates not only dopaminergic fl but also serotonergic neurotransmission. In WT but not Taar1−/− phenyl)-4-pyrrolidin-1-yl-3-tri uoromethyl-benzamide (EPPTB). Use of EPPTB revealed that TAAR1 tonically activates inwardly mice, RO5166017 prevented stress-induced hyperthermia and rectifying K+ channels in VTA DA neurons to reduce the basal blocked dopamine-dependent hyperlocomotion in cocaine-treated firing activity (17). Importantly, EPPTB increased agonist potency at and dopamine transporter knockout mice as well as hyperactivity DA D2 receptors while reducing their desensitization rate, strongly induced by an NMDA antagonist. These results tie TAAR1 to the suggesting a functional link between TAAR1 and D2 receptors. control of monoamine-driven behaviors and suggest anxiolytic- Further elucidation of TAAR1 physiological functions and and antipsychotic-like properties for agonists such as RO5166017, validation of its promising therapeutic potential require selective opening treatment opportunities for psychiatric disorders. agonists. The ideal characteristics of such molecules are (i) high affinity and intrinsic activity (equal or superior to TAs), (ii) high drug discovery | serotonin | depression | schizophrenia | anxiety selectivity for TAAR1 (in contrast to TAs), (iii) low metabolic turnover (in contrast to TAs), and (iv) favorable pharmacokinetic fi properties, such as a good brain penetration, to enable in vivo n 2001, identi cation of the trace amine-associated receptor fi I1 (TAAR1) provided evidence for a direct biological effect of studies. Here, we report the identi cation of such a compound, so-called trace amines (TAs) (1, 2), a subgroup of biogenic amines RO5166017, which was used for the study of TAAR1 function in previously denoted as false neurotransmitters (3). TAs such as vitro and in vivo. p-tyramine (pTyr), β-phenylethylamine (PEA), octopamine, and tryptamine are metabolites of amino acids with structural simi- larity to classical biogenic amines. Although they are only found Author contributions: F.G.R., J.-L.M., R.R.G., M.G.C., and M.C.H. designed research; F.G.R., at low concentrations in the brain, TAs have been implicated in J.-L.M., R.R.G., A.B., T.D.S., R.M., S.D., K.G.Z., R.N., C.A.M., V.M., S.C., L.O., G.T., and M.C.H. a wide range of neuropathological disorders, including schizo- performed research; K.G.Z. and R.N. contributed new reagents/analytic tools; F.G.R., ’ J.-L.M., R.R.G., A.B., T.D.S., R.M., S.D., C.A.M., V.M., S.C., G.T., B.P., B.B., M.G.C., J.G.W., phrenia, major depression, anxiety states, Parkinson s disease, and M.C.H. analyzed data; and F.G.R., J.-L.M., R.R.G., G.T., and M.C.H. wrote the paper. and attention deficit hyperactivity disorder (3, 4). Conflict of interest statement: F.G.R., J.-L.M., R.M., S.D., K.G.Z., R.N., C.A.M., V.M., S.C., TAAR1, a member of the TAAR family (5, 6), is a G protein- L.O., G.T., B.P., J.G.W., and M.C.H. are F. Hoffmann-La Roche employees. R.R.G. is sup- coupled receptor (GPCR) that signals through Gs to elevate ported in part by research grants from F. Hoffmann-La Roche Ltd. (Basel, Switzerland) and intracellular cAMP levels in response to TAs (6, 7). In vitro Compagnia di San Paolo Fondazione (Turin, Italy). A.B. is employed by Neuroservice. M.G.C. studies have shown a reciprocal regulation between TAAR1 and has received funds for Sponsored Research Agreements unrelated to this work from monoaminergic transporters, particularly the dopamine trans- Forest Laboratories, NeuroSearch, and Lundbeck USA as well as consulting fees from porter (DAT) (8, 9). In the mouse brain, Taar1 is expressed Merck and F. Hoffmann-La Roche Ltd. An unrestricted gift to Duke University was pro- throughout the limbic and monoaminergic systems, including the vided by Lundbeck USA to support Neuroscience research in the laboratory of M.G.C. ventral tegmental area (VTA) and dorsal raphe nucleus (DRN) This article is a PNAS Direct Submission. −/− (10). Mice lacking Taar1 (Taar1 mice) have no overt phenotype 1To whom correspondence should be addressed. E-mail: [email protected]. and appear similar to WT littermates in most neurological and −/− This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. behavioral tests (10–12). However, Taar1 mice are hypersen- 1073/pnas.1103029108/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1103029108 PNAS | May 17, 2011 | vol. 108 | no. 20 | 8485–8490 Downloaded by guest on October 1, 2021 Results Identification of RO5166017 and in Vitro Pharmacological Charac- terization. There seems to be considerable overlap between the pharmacophore space occupied by TAAR1 ligands and ligands of other biogenic amine receptors (2). Therefore, a medicinal chemistry program seeking to create TAAR1 ligands starting from Fig. 1. Chemical structure of the selective TAAR1 agonist RO5166017. adrenergic ligands as a source of lead compounds was started. One of the starting compounds was the amino-oxazoline S18616 (18), α an 2A-adrenergic receptor agonist, and an iterative series of Taar1 fi Because is also expressed in the DRN (10), we asked structural modi cations within this chemical class led to the dis- whether RO5166017 and pTyr also affect the firing frequency covery of RO5166017 [(S)-4-[(ethyl-phenyl-amino)-methyl]-4,5- of 5-HT neurons. Both compounds decreased the spike rate of dihydro-oxazol-2-ylamine] (Fig. 1). DRN 5-HT neurons in WT mice (Fig. 2 C and D and Fig. S3), with fi RO5166017 exhibited high potency and ef cacy at mouse, rat, an IC50 value of 2.99 nM for RO5166017 (Table 1). As in the cynomolgus monkey, and human TAAR1 stably expressed in VTA, the inhibitory effects of pTyr and RO5166017 were blocked HEK293 cells (Table 1). Maximal cAMP levels reached by by EPPTB, which increased the firing frequency over the basal RO5166017 stimulation (maximal efficacy) were in a range similar level. The outward current generated by RO5166017 in the DRN to that achieved by PEA stimulation (set as 100%) at human, rat, was also K+-mediated (Fig. S2 C and D). Finally, neither and monkey TAAR1 (81–95%) and somewhat lower at mouse C D fi RO5166017 (Fig. 2 and ) nor−/− pTyr (Fig. S3) affected the ring TAAR1 (65%). Electrophysiological recordings in Xenopus oocytes rate of 5-HT neurons in Taar1 mice, supporting the specificity as well as VTA and DRN slices resulted in EC50 and IC50 values for of their effects in WT mice. Interestingly, the spontaneous spike mouse TAAR1 (1.7–8 nM) comparable with those obtained by the rate in DRN slices from mutant mice was significantly increased cAMP assay in HEK293 cells (3.3 nM).
Recommended publications
  • National Center for Toxicological Research
    National Center for Toxicological Research Annual Report Research Accomplishments and Plans FY 2015 – FY 2016 Page 0 of 193 Table of Contents Preface – William Slikker, Jr., Ph.D. ................................................................................... 3 NCTR Vision ......................................................................................................................... 7 NCTR Mission ...................................................................................................................... 7 NCTR Strategic Plan ............................................................................................................ 7 NCTR Organizational Structure .......................................................................................... 8 NCTR Location and Facilities .............................................................................................. 9 NCTR Advances Research Through Outreach and Collaboration ................................... 10 NCTR Global Outreach and Training Activities ............................................................... 12 Global Summit on Regulatory Science .................................................................................................12 Training Activities .................................................................................................................................14 NCTR Scientists – Leaders in the Research Community .................................................. 15 Science Advisory Board ...................................................................................................
    [Show full text]
  • 'Gating' Residues Ile199 and Tyr326 in Human Monoamine Oxidase B
    The ‘gating’ residues Ile199 and Tyr326 in human monoamine oxidase B function in substrate and inhibitor recognition Erika M. Milczek1,*, Claudia Binda2, Stefano Rovida2, Andrea Mattevi2 and Dale E. Edmondson1 1 Departments of Chemistry and Biochemistry, Emory University, Atlanta, Georgia, USA 2 Department of Genetics and Microbiology, University of Pavia, Italy Keywords The major structural difference between human monoamine oxidases A dipartite to monopartite cavity conversion; (MAO A) and B (MAO B) is that MAO A has a monopartite substrate inhibitor specificity; monoamine oxidase B; cavity of 550 A˚3 volume and MAO B contains a dipartite cavity struc- mutations of gating residues; structure of ture with volumes of 290 A˚3 (entrance cavity) and 400 A˚3 (substrate methylene blue complex cavity). Ile199 and Tyr326 side chains separate these two cavities in MAO Correspondence B. To probe the function of these gating residues, Ile199Ala and Ile199Ala- D. E. Edmondson, Department of Tyr326Ala mutant forms of MAO B were investigated. Structural data on Biochemistry, Emory University, 1510 the Ile199Ala MAO B mutant show no alterations in active site geometries Clifton Road, Atlanta, GA 30322, USA compared with wild-type enzyme while the Ile199Ala-Tyr326Ala MAO B Fax: +1 404 727 2738 mutant exhibits alterations in residues 100–103 which are part of the loop Tel: +1 404 727 5972 gating the entrance to the active site. Both mutant enzymes exhibit catalytic E-mail: [email protected] properties with increased amine KM but unaltered kcat values. The altered *Present address KM values on mutation are attributed to the influence of the cavity struc- Department of Chemistry, Princeton ture in the binding and subsequent deprotonation of the amine substrate.
    [Show full text]
  • Parkinson Disease and Other Movement Disorders
    P1: Trim: 8.375in × 10.875in Top: 0.373in Gutter: 0.664in LWBK915-57 LWW-KodaKimble-educational September 17, 2011 2:31 Parkinson Disease and Other 57 Movement Disorders Michael E. Ernst and Mildred D. Gottwald CORE PRINCIPLES CHAPTER CASES PARKINSON DISEASE 1 Parkinson disease (PD) is a chronic, progressive movement disorder resulting from Case 57-1 (Questions 1, 2) loss of dopamine from the nigrostriatal tracts in the brain, and is characterized by rigidity, bradykinesia, postural disturbances, and tremor. 2 Treatment for PD is aimed at restoring dopamine supply through one, or a Case 57-1 combination, of the following methods: exogenous dopamine in the form of a (Questions 3–18), precursor, levodopa; direct stimulation of dopamine receptors via dopamine Case 57-2 (Questions 1, 2) agonists; and inhibition of metabolic pathways responsible for degradation of levodopa. 3 Therapy for PD is usually delayed until there is a significant effect on quality of life; Case 57-1 (Questions 4, 10) generally younger patients start with dopamine agonists, whereas older patients may start with levodopa. 4 Initial therapy with dopamine agonists is associated with a lower risk of developing Case 57-1 (Questions 4, motor complications than with levodopa, but all patients will eventually require 10–15) levodopa. 5 Advanced PD is characterized by motor fluctuations including a gradual decline in Case 57-1 (Questions on time, and the development of troubling dopaminergic-induced dyskinesias. 15–18), Case 57-2 Dopamine agonists, monoamine oxidase type B (MAO-B) inhibitors, and (Question 1), Case 57-3 catechol-O-methyltransferase (COMT) inhibitors can reduce motor fluctuations; (Questions 1, 2) amantadine can improve dyskinesias.
    [Show full text]
  • Recent Applications of Capillary Electrophoresis in the Determination of Active Compounds in Medicinal Plants and Pharmaceutical Formulations
    molecules Review Recent Applications of Capillary Electrophoresis in the Determination of Active Compounds in Medicinal Plants and Pharmaceutical Formulations Marcin Gackowski 1,* , Anna Przybylska 1 , Stefan Kruszewski 2 , Marcin Koba 1 , Katarzyna M ˛adra-Gackowska 3 and Artur Bogacz 4 1 Department of Toxicology and Bromatology, Faculty of Pharmacy, L. Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, A. Jurasza 2 Street, PL–85089 Bydgoszcz, Poland; [email protected] (A.P.); [email protected] (M.K.) 2 Biophysics Department, Faculty of Pharmacy, L. Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Jagiello´nska13 Street, PL–85067 Bydgoszcz, Poland; [email protected] 3 Department of Geriatrics, Faculty of Health Sciences, L. Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Skłodowskiej Curie 9 Street, PL–85094 Bydgoszcz, Poland; [email protected] 4 Department of Otolaryngology and Oncology, Faculty of Medicine, L. Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Skłodowskiej Curie 9 Street, PL–85094 Bydgoszcz, Poland; [email protected] Citation: Gackowski, M.; Przybylska, * Correspondence: [email protected] A.; Kruszewski, S.; Koba, M.; M ˛adra-Gackowska,K.; Bogacz, A. Abstract: The present review summarizes scientific reports from between 2010 and 2019 on the use Recent Applications of Capillary of capillary electrophoresis to quantify active constituents (i.e., phenolic compounds, coumarins, Electrophoresis in the Determination protoberberines, curcuminoids, iridoid glycosides, alkaloids, triterpene acids) in medicinal plants and of Active Compounds in Medicinal herbal formulations. The present literature review is founded on PRISMA guidelines and selection Plants and Pharmaceutical criteria were formulated on the basis of PICOS (Population, Intervention, Comparison, Outcome, Formulations.
    [Show full text]
  • Β-Carboline Alkaloids and Essential Tremor: Exploring the Environmental Determinants of One of the Most Prevalent Neurological Diseases
    Review TheScientificWorldJOURNAL (2010) 10, 1783–1794 ISSN 1537-744X; DOI 10.1100/tsw.2010.159 β-Carboline Alkaloids and Essential Tremor: Exploring the Environmental Determinants of One of the Most Prevalent Neurological Diseases Elan D. Louis1,2,3,4,* and Wei Zheng5 1GH Sergievsky Center, 2Department of Neurology, 3Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, College of Physicians and Surgeons, Columbia University, New York; 4Department of Epidemiology, Mailman School of Public Health, Columbia University, New York; 5School of Health Sciences, Purdue University, West Lafayette, IN E-mail: [email protected]; [email protected] Received May 24, 2010; Revised July 15, 2010; Accepted July 15, 2010; Published September 1, 2010 Essential tremor (ET) is among the most prevalent neurological diseases, yet its etiology is not well understood. Susceptibility genotypes undoubtedly underlie many ET cases, although no genes have been identified thus far. Environmental factors are also likely to contribute to the etiology of ET. Harmane (1-methyl-9H-pyrido[3,4- β]indole) is a potent, tremor-producing β-carboline alkaloid, and emerging literature has provided initial links between this neurotoxin and ET. In this report, we review this literature. Two studies, both in New York, have demonstrated higher blood harmane levels in ET cases than controls and, in one study, especially high levels in familial ET cases. Replication studies of populations outside of New York and studies of brain harmane levels in ET have yet to be undertaken. A small number of studies have explored several of the biological correlates of exposure to harmane in ET patients.
    [Show full text]
  • MALDI Mass Spectrometry Imaging for Visualizing in Situ Metabolism of Endogenous Metabolites and Dietary Phytochemicals
    Metabolites 2014, 4, 319-346; doi:10.3390/metabo4020319 OPEN ACCESS metabolites ISSN 2218-1989 www.mdpi.com/journal/metabolites/ Review MALDI Mass Spectrometry Imaging for Visualizing In Situ Metabolism of Endogenous Metabolites and Dietary Phytochemicals Yoshinori Fujimura * and Daisuke Miura * Innovation Center for Medical Redox Navigation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan * Authors to whom correspondence should be addressed; E-Mails: [email protected] (Y.F.); [email protected] (D.M.); Tel.: +81-92-642-6160 (Y.F.); +81-92-642-6091 (D.M.); Fax: +81-92-642-6285 (Y.F.); +81-92-642-6285 (D.M.). Received: 26 February 2014; in revised form: 17 April 2014 / Accepted: 4 May 2014 / Published: 5 May 2014 Abstract: Understanding the spatial distribution of bioactive small molecules is indispensable for elucidating their biological or pharmaceutical roles. Mass spectrometry imaging (MSI) enables determination of the distribution of ionizable molecules present in tissue sections of whole-body or single heterogeneous organ samples by direct ionization and detection. This emerging technique is now widely used for in situ label-free molecular imaging of endogenous or exogenous small molecules. MSI allows the simultaneous visualization of many types of molecules including a parent molecule and its metabolites. Thus, MSI has received much attention as a potential tool for pathological analysis, understanding pharmaceutical mechanisms, and biomarker discovery. On the other hand, several issues
    [Show full text]
  • EFFECTS of FLAVONOIDS and OTHER PHYTOCHEMICALS on FISH Cypla MONOOXYGENASES, EMBRYONIC and REPRODUCTIVE DEVELOPMENT
    EFFECTS OF FLAVONOIDS AND OTHER PHYTOCHEMICALS ON FISH CYPlA MONOOXYGENASES, EMBRYONIC AND REPRODUCTIVE DEVELOPMENT A Thesis Submitted to the Committee of Graduate Studies in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the Faculty of Arts and Science Trent University Peterborough. Ontario. Canada 0 Y iannis Kiparissis 200 1 Watershed Ecosystems Ph.D. Program June 2001 National Library Bibliothbque nationale du Canada Acquisitions and Acquisitions et Bibliographic Sewices services bibliographiques 395 Wellington Street 395, rue WeUington Ottawa ON KIA ON4 Ottawa ON KIAON4 Canada Canada The author has granted a non- L'auteur a accorde une Licence non exclusive licence allowing the exclusive pernettant a la National Library of Canada to Bibliotheque nationale du Canada de reproduce, loan, distr!iiute or sell reproduire, preter, distniuer ou copies of this thesis in microform, vendre des copies de cette these sous paper or electronic formats. la forme de microfiche/film, de reproduction sur papier ou sur format electronique. The author retains ownership of the L'auteur conserve la propriete du copyright in this thesis. Neither the droit d'auteur qui protege cette these. thesis nor substantial extracts &om it Ni la these ni des extraits substantiels may be printed or othenvise de celfe-ci ne doivent &re imprimes reproduced without the author's ou autrement reproduits sans son permission. autorisation. ABSTRACT Effects of Flavonoids and Other Phytochemicals on Fish CYPIA Monooxygenases, Embryonic and Reproductive Development The purpose of this study was to demonstrate that biological responses in fish such as induction of CYP1 A-dependent monooxygenases. embryonic defects.
    [Show full text]
  • Download Product Insert (PDF)
    PRODUCT INFORMATION Harmane Item No. 29613 CAS Registry No.: 486-84-0 Formal Name: 1-methyl-9H-pyrido[3,4-b]indole Synonyms: 1-Methyl-β-Carboline, NSC 54439 H MF: C H N 12 10 2 N FW: 182.2 Purity: ≥98% N UV/Vis.: λmax: 213, 234, 249, 288 nm Supplied as: A solid Storage: -20°C Stability: ≥2 years Information represents the product specifications. Batch specific analytical results are provided on each certificate of analysis. Laboratory Procedures Harmane is supplied as a solid. A stock solution may be made by dissolving the harmane in the solvent of choice, which should be purged with an inert gas. Harmane is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide (DMF). The solubility of harmane in ethanol is approximately 10 mg/ml and approximately 20 mg/ml in DMSO and DMF. Harmane is sparingly soluble in aqueous buffers. For maximum solubility in aqueous buffers, harmane should first be dissolved in DMSO and then diluted with the aqueous buffer of choice. Harmane has a solubility of approximately 0.25 mg/ml in a 1:3 solution of DMSO:PBS (pH 7.2) using this method. We do not recommend storing the aqueous solution for more than one day. Description Harmane is a β-carboline that has been found in P. harmala, as well as in cooked meats and tobacco 1-7 and has diverse biological activities. It is an inhibitor of monoamine oxidase A (MAO-A; IC50 = 0.5 µM) 3 that also inhibits MAO-B (IC50 = 5 µM).
    [Show full text]
  • Gaurtieraa 1939Redux.Pdf (9.693Mb)
    SYNTHESIS AND RESOLUTION OF PUKATEINE AND OF A CLOSELY RELATED ALKALOID. by Alec A. Gautier (Geneva) Dipl. Ing. E.T.H. Zürich. Thesis submitted for the Degree of Ph.D. University of Edinburgh. April 1939. Preface. This thesis embodies the results of synthetical work carried out in the laboratory of the Medical Chemistry Department of the University of Edinburgh during the academic years 1°37 -38 and 193M -39, under the supervision of the late Professor G. Barger, F.R.S. The author wishes to express his sincere thanks to Professor G.F. Marrian of the Medical Chemistry Department of the University of Edinburgh, for his valuable advice towards the competion of this work, and to the Moray Fund of the University of Edinburgh for a grant which has defrayed the cost of materials. Contents. Page I. THEORY OF THE ORIGIN OF ALKALOIDS.. 1 II. SYNTHESIS OF PUKATEINE. (a) Theoretical 21 (b) Experimental .. .. .. .. .. 37 III. SYNTTHESIS AND RESOLUTION OF A CLOSELY RELATED ALKALOID. (a) Theoretical . 52 (b) Experimental .. .. .. .. .. 56 References .. .. 72 -1- I. THEORY OF THE ORIGIN OF ALKALOIDS. The wide occurrence of alkaloids in the vegetable kingdom makes it reasonable to assume that they must fulfil some important function in the life of the plant. Although this matter has often been dis- cussed and investigated experimentally, it cannot be said that the solution of the problem is in sight. Heckel put forward the assumption that the alkaloids were intermediate products in the building up of protoplasm. If this is the case at all, it must be confined to a few alkaloids which are related to the protein amino- acids.
    [Show full text]
  • Hyperkinetic Movement Disorders Differential Diagnosis and Treatment
    Hyperkinetic Movement Disorders Differential diagnosis and treatment Albanese_ffirs.indd i 1/23/2012 10:47:45 AM Wiley Desktop Edition This book gives you free access to a Wiley Desktop Edition – a digital, interactive version of your book available on your PC, Mac, laptop or Apple mobile device. To access your Wiley Desktop Edition: • Find the redemption code on the inside front cover of this book and carefully scratch away the top coating of the label. • Visit “http://www.vitalsource.com/software/bookshelf/downloads” to download the Bookshelf application. • Open the Bookshelf application on your computer and register for an account. • Follow the registration process and enter your redemption code to download your digital book. • For full access instructions, visit “http://www.wiley.com/go/albanese/movement” Companion Web Site A companion site with all the videos cited in this book can be found at: www.wiley.com/go/albanese/movement Albanese_ffirs.indd ii 1/23/2012 10:47:45 AM Hyperkinetic Movement Disorders Differential diagnosis and treatment EDITED BY Alberto Albanese MD Professor of Neurology Fondazione IRCCS Istituto Neurologico Carlo Besta Università Cattolica del Sacro Cuore, Milan, Italy Joseph Jankovic MD Professor of Neurology Director, Parkinson’s Disease Center and Movement Disorders Clinic Department of Neurology Baylor College of Medicine Houston, TX, USA A John Wiley & Sons, Ltd., Publication Albanese_ffirs.indd iii 1/23/2012 10:47:45 AM This edition first published 2012, © 2012 by Blackwell Publishing Ltd Blackwell Publishing was acquired by John Wiley & Sons in February 2007. Blackwell’s publishing program has been merged with Wiley’s global Scientific, Technical and Medical business to form Wiley-Blackwell.
    [Show full text]
  • Exploring the Physiological Role of Vibrio Fischeri Pepn
    EXPLORING THE PHYSIOLOGICAL ROLE OF VIBRIO FISCHERI PEPN A thesis presented to the Faculty of California Polytechnic State University, San Luis Obispo In Partial Fulfillment of the Requirements for the Degree Master of Science in Biological Sciences by Sally L. Cello April 2015 © 2015 Sally L. Cello ALL RIGHTS RESERVED ii COMMITTEE MEMBERSHIP TITLE: Exploring the physiological role of Vibrio fischeri PepN AUTHOR: Sally L. Cello DATE SUBMITTED: April 2015 COMMITTEE CHAIR: Pat Fidopiastis, Ph.D. Associate Professor of Biological Sciences COMMITTEE MEMBER: Chris Kitts, Ph.D. Department Chair of Biological Sciences Professor of Biological Sciences COMMITTEE MEMBER: Candace Winstead, Ph.D. Associate Professor of Biological Sciences iii ABSTRACT Exploring the physiological role of Vibrio fischeri PepN Sally L. Cello The primary contributor to Vibrio fischeri aminopeptidase activity is aminopeptidase N, PepN. Colonization assays revealed the pepN mutant strain to be deficient at forming dense aggregates and populating the host’s light organ compared to wildtype within the first 12 hours of colonization; however the mutant competed normally at 24 hours. To address the role of PepN in colonization initiation and establish additional phenotypes for the pepN mutant strain, stress response and other physiological assays were employed. Marked differences were found between pepN mutant and wildtype strain in response to salinity, acidity, and antibiotic tolerance. This study has provided a foundation for future work on identifying a putative role for V. fischeri PepN in regulating stress response. iv ACKNOWLEDGMENTS Thank you to my wonderful friends and family for their endless support during this journey. Thank you to Pat Fidopiastis for his guidance and quick replies! Thank you to my committee members for their words of wisdom contributing to a well-polished paper.
    [Show full text]
  • View Preprint
    A peer-reviewed version of this preprint was published in PeerJ on 22 June 2016. View the peer-reviewed version (peerj.com/articles/2130), which is the preferred citable publication unless you specifically need to cite this preprint. Read HM, Mills G, Johnson S, Tsai P, Dalton J, Barquist L, Print CG, Patrick WM, Wiles S. 2016. The in vitro and in vivo effects of constitutive light expression on a bioluminescent strain of the mouse enteropathogen Citrobacter rodentium. PeerJ 4:e2130 https://doi.org/10.7717/peerj.2130 The in vitro and in vivo effects of constitutive light expression on the mouse enteropathogen Citrobacter rodentium Hannah M Read, Grant Mills, Sarah Johnson, Peter Tsai, James Dalton, Lars Barquist, Cristin G Print, Wayne M Patrick, Siouxsie Wiles Bioluminescent reporter genes, such as those from fireflies and bacteria, let researchers use light production as a non-invasive and non-destructive surrogate measure of microbial numbers in a wide variety of environments. As bioluminescence needs microbial metabolites, tagging microorganisms with luciferases means only live metabolically active cells are detected. Despite the wide use of bioluminescent reporter genes, very little is known about the impact of continuous (also called constitutive) light expression on tagged bacteria. We have previously made a bioluminescent strain of Citrobacter rodentium, a bacterium which infects laboratory mice in a similar way to how enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC) infect humans. In this study, we investigated whether constitutive light expression makes the bioluminescent C. rodentium strain ICC180 less competitive when competed against its non-bioluminescent parent (strain ICC169).
    [Show full text]