Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-159004 from National Institute On Drug Abuse
Project start date: 2001-08-01
Project end date: 2002-09-29
Sponsored Links Excellgen http://Excellgen.com
ROLE OF INTERNALIZATION IN THE FUNCTIONAL REGULATION OF OPIOID And ORL-1 RECEPTORS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-130009 from National Institute On Drug Abuse
Abstract: To study the trafficking of mu, delta and ORL-1 receptors and their ligands following receptor activation. a). What is the cellular distribution and/or vesicular localization of mu receptors both in vitro and in vivo following various acute and chronic agonist treatments. In vitro, is the trafficking of the delta and ORL-1 receptors similar to that of the mu receptor? b) With the development and use of fluorescent opioid peptide ligands, can we study the trafficking of ligands and receptors be observed in real time. C) What are the kinetics of recover of surface opioid and ORL-1 receptors following agonist treatment both in vitro and in vivo? What are the processes involved in the recovery of surface receptor in vitro. 2. To identify the structural features of opioid ligands which are critical for triggering mu, delta,, and ORL-1 receptor sequestration and determine if any correlation exists between a ligand s ability to trigger internalization and its potency or efficacy for other receptor functions. a) What structural features of peptide and alkaloid ligands are required to induce sequestration of mu, delta and ORL-1 receptors in vitro. b) What is the relationship between a ligand s ability to trigger internalization and its binding affinity potency or efficacy in inhibition of cAMP accumulation and stimulation of GTPgammaS binding in mu, delta and ORL-1 receptors. c) Is an agonist s capacity to cause internalization the same in vivo as in vitro for mu, delta and ORL-1 receptors? 3. To determine the functional significance of sequestration of mu receptors. a. Is there a difference in desensitization or resensitization of the mu receptor in vitro between agonists that induce sequestration verses those that do not? b. Do mu agonists that induce internalization have the same effects on long-term potentiation (LTP) as those which do not? c. How do adaptational processes differ between wild type and internalization-deficient mutant mu receptors? d. What are the behavioral manifestations and pharmacological consequences of opioid treatment in a mouse expressing an internalization-deficient mu receptor? 4. To determine whether mu, delta, or ORL-1 receptor internalization can be observed under conditions which induce the endogenous release of opioid peptides. a. Can receptor sequestration be observed in the CNS following stress- induced analgesia (SIA) using the swim stress model? b. Does amygdala kindling result in receptor sequestration? c. Does stimulation of the perforant pathway of the hippocampus result in receptor sequestration?
Keywords: opioid receptor, protein structure /function, receptor binding, receptor expression, ligand, neuropeptide, opioid, protein transport, autoradiography, fluorescent dye /probe, laboratory mouse, molecular cloning, peptide chemical synthesis, transgenic animal, video microscopy
CENTER FOR STUDY OF NEUROREGULATORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-06 from National Institute On Drug Abuse IRG: SRCD
Abstract: This NIDA Center Grant application is for studies of the biomedical aspects of drug abuse with a primary focus on ligand- receptor processes. The central goal is to use the tools of molecular biology and peptide chemistry to study key processes of the opioid systems including their ligands, receptors, and processing enzymes in a coordinated manner to investigate the differential, region-specific and time-related effects of drugs of abuse. The research ultimately will permit the development of human probes for clinical investigation of differential diagnosis, treatment, and prevention. The research is oriented to the mechanisms underlying drug tolerance and ultimately addiction and withdrawal with special emphasis on the endorphins, or opioid peptides, naturally present in nervous tissue which have a key role in drug abuse. The specific goals are to study 1) opioid precursors in terms of their processing and regulation in order to determine the ways in which the naturally present substances are controlled and how they are affected by drugs of abuse; 2) enzymes which process opioid peptides by cloning them and investigating their normal regulation and how they are altered by drugs of abuse; and 3) the receptors of the opioids by cloning them and investigating their regulation since the receptors may be key to understanding the effects of drugs. As parts of the Center, other drug abuse systems will also be examined. The proposed program expands ongoing studies of members of the Center using the techniques of protein and peptide chemistry (including peptide synthesis, immunoassays, peptide and protein isolation); molecular biology (including cDNA cloning and mRNA techniques); histochemistry (including light and EM immunocytochemistry and in situ hybridization); analytical neurochemistry (including fast atom bombardment mass spectrometry); and human studies in a cross-disciplinary and integrated manner. The more precise understanding of the mechanisms of tolerance, addiction, and withdrawal yielded by the studies and the probes should provide the basis for improving methods of diagnosis, treatment, and prevention of drug abuse.
Project start date: 1987-09-30
Project end date: 1993-07-31
5P50DA005010-06 (1992): $672296
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-09 from National Institute On Drug Abuse IRG: SRCD
Abstract: This continuation of the NlDA Center at UCLA will further characterize key molecules and mechanisms involved in opioid receptor regulation with the broad goal of more clearly understanding the processes of tolerance, dependence and withdrawal associated with drugs of abuse. This basic research ultimately will permit the development of human probes and reagents for clinical investigations addressing differential diagnosis and treatment. The focus of the continuing research of the Center is based primarily upon our successful isolation of a delta opioid receptor gene (DOR-1) and draws upon the breadth of opioid pharmacological and neurochemical accomplishments of the Center. The program comprises five major research areas 1) The characterization of gene(s) and mRNAs encoding DOR-1. 2) The determination of the structural basis for opiate alkaloid and opioid peptide recognition and for signal transduction. 3) The identification and structural characterization of opioid receptors related to, but distinct from, DOR-1. 4) The analysis of opioid receptor neuroanatomical localization and relationship with other neurotransmitter systems implicated in drug abuse. 5) The understanding of mRNA regulation, post-translational modifications and intracellular trafficking of opioid receptors in both normal and drug perturbed states. The resources and expertise incorporated in the Center span techniques of molecular biology (cDNA cloning, mRNA analysis, genomic analysis, mutagenesis and expression), protein and peptide chemistry (peptide synthesis, peptide and protein purification, protein sequencing), analytical neurochemistry (mass spectroscopy, immunoassay, amino acid analysis), neuropharmacology (receptor binding, second messenger systems, microdialysis, behavior), and histochemistry (immunocytochemistry and in situ hybridization). Functionally the Center s research areas are highly integrated, interactive and interdependent. The continued application of this successful multidisciplinary and collaborative approach will lead to a more precise understanding of the molecular mechanisms underlying the addictive process and thus provide a basis for improved clinical methods for ameliorating the problems associated with drugs of abuse.
Keywords: neurotransmitter, opiate alkaloid, opioid receptor
Project start date: 1987-09-30
Project end date: 1997-07-31
5P50DA005010-09 (1995): $644166
5P50DA005010-08 (1994): $704868
5P50DA005010-20 (2006): $1653580
5P50DA005010-19 (2005): $1647091
5P50DA005010-18 (2004): $1417340
5P50DA005010-17 (2003): $1376056
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Abstract: The Administrative Core provides programmatic structure, logistical support, and mediation of the Center´s interactions with the training programs, administrative offices, and scientific and governmental institutions. The Core also is responsible for overseeing the Pilot Program. Day-to-day operation of the Core is the responsibility of the Core Supervisor (Terry Novorr), with immediate oversight by the Administrative Core Director, Dr. Chris Evans or in his absence, the Associate Director Dr. Nigel Maidment. The Core coodinates many educational functions of the center, including its translational efforts spearheaded by Dr Martin Iguchi. Responsibilities of the Core include ¿ Coordination and evaluation of the Center´s scientific directions, educational roles, progress and interactions. ¿ Overseeing operation of the Pilot Program; including advertising for potential grantees, evaluation of proposals and integration with ongoing research in CSORDA. ¿ Resolving personnel issues including hiring, conflicts, promotions and replacements. ¿ Logistical support for publications, grant preparations, coordinating of meetings, travel and purchasing. ¿ Budgetary preparation, prioritization and monitoring. The Administrative Core is essential to the operation of CSORDA
Keywords: Address; Administrator; Advertisements; Advertising; Animal Experimental Use; Animal Experimentation; Animal Housing; Animal Research; Animals; Applications Grants; Award; Book Chapters; Budgets; Carbon Dioxide Snow; Cell Isolation; Cell Segregation; Cell Separation; Cell Separation Technology; Charge; Chemicals; Computer Programs; Computer software; Computers; Conflict; Conflict (Psychology); Contracting Opportunities; Contracts; Dry Ice; Ensure; Environment; Equipment; Equipment and supply inventories; Evaluation; Expenditure; Faculty; Fees; Fellowship; Funding; Funding Opportunities; Gases; Grant; Grant Proposals; Grants, Applications; Guidelines; Human Resources; Institution; Interview; Inventory; Journals; Laboratories; Lectures; Lectures (PT); Lectures [Publication Type]; Libraries; Licensing; Magazine; Mails; Maintenance; Maintenances; Manpower; Manuscripts; Mediation; Mentors; Microscope; Monitor; Negotiating; Negotiation; Neurosciences; Office Management; Operation; Operative Procedures; Operative Surgical Procedures; PYMT; Participant; Peptide Synthesis; Personnel Staffing; Pilot Projects; Policies; Preparation; Procedures; Process; Program Evaluation; Programs (PT); Programs [Publication Type]; Progress Reports; Publications; Publishing; Radiation; Radioactive; Reagent; Recommendation; Refuse Disposal; Refuse Disposals; Research; Research Resources; Resolution; Resources; Role; SCHED; Safety; Schedule; Scientific Publication; Services; Software; Speech; Structure; Students; Substance abuse problem; Surgical; Surgical Interventions; Surgical Procedure; Training; Training Activity; Training Programs; Travel; Universities; Update; Visit; Work; ing; abuse of substances; animal care; cell sorting; computer network; computer program/software; graduate student; lectures; medical schools; meetings; member; payment; personnel; pilot study; programs; ray (radiation); response; satisfaction; social role; substance abuse; surgery; waste disposal
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
5P50DA005010-24_9010 (2010): $145978
Sponsored Links Excellgen http://Excellgen.com
OPIOID RECEPTOR SIGNALING: SELECTIVE MECHANISM OF REGULATION & RECEPTOR CROSSTALK
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Abstract: Principal Investigator/Program Director Evans, Christopher J., Ph.D., Component II Component II title Opioid Receptor Signaling selective mechanisms of regulation and receptor cross-talk Component II will study the mechanisms by which mu opioid agonists can trigger differential signaling and receptor trafficking. During the past funding period we have developed mouse dorsal root ganglia (DRG) primary cultures to study both native and virally expressed opioid receptors in wild-type and mutant mice (such as those generated in Component I and others available through the Mutant Mouse Core). The use of real time PCR, gene arrays, flow cytometry and confocal microscopy has enabled us to fully characterize the expression of regulated mRNAs as well as protein expression, distribution, and states of phosphorylation. Using these techniques in combination with electrophysiological recording, we have developed evidence for unique trafficking and signaling profiles for morphine and DAMGO with implication for a major regulatory Vole of arrestin isoforms. Based upon these preliminary data Component II will 1) Characterize the differential mu-receptor-mediated signaling initially observed with DAMGO and morphine and subsequently define signaling profiles of other clinically important and endogenous agonists; 2) Analyze the contributions of receptor cross-talk via alpha 2a and delta opioid receptors on mu-receptor trafficking and signaling; 3) Test the hypothesis that beta-arrestins, and the cellular targeting of c-src are critical in determining both the opioid agonist signaling-profiles and trafficking of mu opioid receptors. Together these aims will provide a foundation for understanding agonist-directed signaling via mu opioid receptors with the goal of identifying agonist properties that may differentiate the clinically useful from the detrimental effects of opioid drugs
Keywords: 3-Hepta, 6-(dimethylamino)-4, 4-diphenyl-; 6, 14-Ethenomorphinan-7-methanol, 17-(cyclopropylmethyl)-alpha-(1, 1-dimethylethyl)-4, 5-epoxy-18, 19-dihydro-3-hydroxy-6-methoxy-alpha-methyl-, (5alpha, 7alpha(S))-; 6, 14-Ethenomorphinan-7-methanol, 4, 5-epoxy-3-hydroxy-6-methoxy-alpha, 17-dimethyl-alpha-propyl-, (5alpha, 7alpha(R))-; ARRB2; ASV; ASVSRC1; Absence of pain sensation; Absence of sensibility to pain; Adanon; Agonist; Ala(2)-MePhe(4)-Gly-ol(5) Enkephalin; Alkaloids; Althose; Arrestins; Behavioral; Buprenorphine; Cell Communication and Signaling; Cell Signaling; Cells; Chronic; Confocal Microscopy; Contin, MS; Cytofluorometry, Flow; D-Ala(2)-MePhe(4)-Gly-ol(5) Enkephalin; D-Ala2-NMe-Phe4-Gly-ol Enkephalin; DAGO; DAGOL; DAMGE; DAMGO; Data; Doctor of Philosophy; Dolophine; Dorsal Root Ganglia; Drugs; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, alanyl(2)-methylphenylalanyl(4)-glycine(5)-; Ethorphine; Etorphine; Exhibits; Extracellular Signal-Regulated Kinase Gene; Feels no pain; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Foundations; Funding; G-Proteins; GTP-Binding Proteins; GTP-Regulatory Proteins; Ganglia, Spinal; Gene Expression; Genes; Genes, c-src; Genetic; Goals; Guanine Nucleotide Coupling Protein; Guanine Nucleotide Regulatory Proteins; Infumorph; Intracellular Communication and Signaling; Investigators; Isoforms; Kadian; L-Phenylalaninamide, L-tyrosyl-D-alanylglycyl-N-(2-hydroxyethyl)-nalpha-methyl-; Ligands; MAP Kinase Gene; MAPK; MSir; Mammals, Mice; Mediating; Medication; Methadone; Methadose; Methods and Techniques; Methods, Other; Mice; Mice, Mutant Strains; Microfluorometry, Flow; Microtus; Mitogen-Activated Protein Kinase Gene; Morphia; Morphinan-3, 6-diol, 7, 8-didehydro-4, 5-epoxy-17-methyl- (5alpha, 6alpha)-; Morphine; Murine; Mus; Mutant Strains Mice; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; Neurons; No sensitivity to pain; Opioid; Opioid Receptor; Oramorph; Oramorph SR; Ph.D.; PhD; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphorylation; Principal Investigator; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Protein Isoforms; Protein Phosphorylation; Receptor Cross-Talk; Receptor Protein; Receptor Signaling; Receptors, Opiate; Receptors, Opioid, delta; Receptors, Opioid, mu; Receptors, delta; Receptors, mu; Regulation; Research Personnel; Researchers; Role; Roxanol; SRC; SRC gene; SRC1; Series; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Spinal Ganglia; Statex SR; System; System, LOINC Axis 4; Techniques; Testing; Time; Transcript; Tyr-Ala-Gly-(NMe)Phe-Gly-ol; V (voltage); Vole; ing; analgesia; base; beta-arrestin; beta-arrestin 2; biological signal transduction; c src; c-src Proto-Oncogenes; cellular targeting; desensitization; dorsal root ganglion; drug/agent; field mouse; flow cytophotometry; inhibitor; inhibitor/antagonist; mouse mutant; neuronal; programs; protein expression; receptor; receptor internalization; receptor-mediated signaling; response; social role; trafficking; v-SRC Avian Sarcoma (Schmidt-Ruppin A-2) Viral Oncogene Homolog; voltage
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
5P50DA005010-24_0016 (2010): $256858
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE (CSORDA)
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Grant 5P50DA005010-24 from National Institute On Drug Abuse
Abstract: Center for study of opioid receptors and drugs of abuse (CSORDA) the research objectives of CSORDA are to gain insights into the mechanisms of action of endogenous opioids and opioid drugs at their cognate receptors with the goal of discerning fundamental processes contributing to behaviors such as analgesia, addiction, tolerance and withdrawal. The Center has four integrated components, which will investigate the activity of opioid ligands at the molecular, cellular and behavioral levels utilizing complementary methodologies and shared resources. The Components of CSORDA will specifically investigate I) The regulation of trafficking and signaling of mu and delta receptors in vivo by high resolution imaging of mice expressing fluorescent receptors; II) The basis for mu ligand- directed signaling and receptor trafficking using primary dorsal root ganglia cells from wild-type as well as receptor and arrestin-deficient mice; III) The role of the endogenous opioid system in goal-directed and habitual behaviors both in drug free and opioid dependent conditions; IV) The functional regulation of transcripts in the striatonigral and striatopallidal projection neuron circuits and their relation to opioid drug reward. In addition to the research specified within the components, a Pilot Program will continue to create new ´ avenues of investigation within the Center and enrich the ongoing programs. For facilitation of the component and pilot projects, an Administrative Core and the CSORDA Mouse Core (CMC), serve to integrate resources and coordinate the needs of the center. The overall mission of CSORDA is to enhance the understanding of the mammalian opioid system thereby providing strategies for improving the clinical use of opioids and treatments for addition
Project start date: 1987-09-30
Project end date: 2012-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
5P50DA005010-24 (2010): $1202180
5P50DA005010-23 (2009): $1178928
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-15 from National Institute On Drug Abuse IRG: ZDA1
Abstract: Applicant s ) The research objectives of the Center for Study of Opioid Receptors and Drugs of Abuse are to gain insights into the mechanisms of action of opioid drugs and their receptors with the ultimate goal of discerning molecular processes that contribute to opiate addiction, tolerance and withdrawal. The Center has four integrated components which have evolved since the cloning of the opioid and ORL-1 receptors. These components will investigate the actions and regulation of receptors both in vitro and in vivo utilizing overlapping methodologies and resources. The Center will specifically investigate 1) The coupling of opioid and ORL-1 receptors through G-proteins and second messenger systems to the release process, with an emphasis on the contribution of specific calcium channel subtypes. 2) The role of receptor internalization in the regulation of receptors and the functional consequences of drugs that induce sequestration (i.e. enkephalins) versus those that do not (i.e. morphine). 3) The role of ORL-1 receptor ligands and their interaction with opiate drugs in the modulation of hippocampal function. 4) Whether ORL-1 receptor agonists have "anti-opioid" effects on brain reward systems. To facilitate the research, the Center has an Administrative Core and five Scientific Cores which serve to integrate its resources, as well as provide practical expertise in specific technical areas. The Scientific Cores cover Molecular Biology, Peptide Chemistry, Analytical Neurochemistry, Tissue Culture and Imaging of both antibody and nucleic acid probes. The continued application of this multidisciplinary and collaborative approach is enhancing our understanding of the molecular mechanisms underlying the addiction process and thus provides a basis for improved clinical methods for ameliorating the problems associated with drugs of abuse.
Keywords: neuropeptide, opiate alkaloid, opioid, opioid receptor
Project start date: 1987-09-30
Project end date: 2002-09-29
5P50DA005010-15 (2001): $1146251
5P50DA005010-14 (2000): $1210262
5P50DA005010-13 (1999): $1112462
5P50DA005010-12 (1998): $1073613
OPIOID RECEPTOR SIGNALING: SELECTIVE MECHANISM OF REGULATION & RECEPTOR CROSSTALK
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Keywords: 3-Hepta, 6-(dimethylamino)-4, 4-diphenyl-; 6, 14-Ethenomorphinan-7-methanol, 17-(cyclopropylmethyl)-alpha-(1, 1-dimethylethyl)-4, 5-epoxy-18, 19-dihydro-3-hydroxy-6-methoxy-alpha-methyl-, (5alpha, 7alpha(S))-; 6, 14-Ethenomorphinan-7-methanol, 4, 5-epoxy-3-hydroxy-6-methoxy-alpha, 17-dimethyl-alpha-propyl-, (5alpha, 7alpha(R))-; ARRB2; ASV; ASVSRC1; Absence of pain sensation; Absence of sensibility to pain; Adanon; Agonist; Ala(2)-MePhe(4)-Gly-ol(5) Enkephalin; Alkaloids; Althose; Arrestins; Behavioral; Buprenorphine; Cell Communication and Signaling; Cell Signaling; Cells; Chronic; Confocal Microscopy; Contin, MS; Cytofluorometry, Flow; D-Ala(2)-MePhe(4)-Gly-ol(5) Enkephalin; D-Ala2-NMe-Phe4-Gly-ol Enkephalin; DAGO; DAGOL; DAMGE; DAMGO; Data; Doctor of Philosophy; Dolophine; Dorsal Root Ganglia; Drugs; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, alanyl(2)-methylphenylalanyl(4)-glycine(5)-; Ethorphine; Etorphine; Exhibits; Extracellular Signal-Regulated Kinase Gene; Feels no pain; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Foundations; Funding; G-Proteins; GTP-Binding Proteins; GTP-Regulatory Proteins; Ganglia, Spinal; Gene Expression; Genes; Genes, c-src; Genetic; Goals; Guanine Nucleotide Coupling Protein; Guanine Nucleotide Regulatory Proteins; Infumorph; Intracellular Communication and Signaling; Investigators; Isoforms; Kadian; L-Phenylalaninamide, L-tyrosyl-D-alanylglycyl-N-(2-hydroxyethyl)-nalpha-methyl-; Ligands; MAP Kinase Gene; MAPK; MSir; Mammals, Mice; Mediating; Medication; Methadone; Methadose; Methods and Techniques; Methods, Other; Mice; Mice, Mutant Strains; Microfluorometry, Flow; Microscopy, Confocal; Microtus; Mitogen-Activated Protein Kinase Gene; Morphia; Morphinan-3, 6-diol, 7, 8-didehydro-4, 5-epoxy-17-methyl- (5alpha, 6alpha)-; Morphine; Murine; Mus; Mutant Strains Mice; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; Neurons; No sensitivity to pain; Opioid; Opioid Receptor; Oramorph; Oramorph SR; Ph.D.; PhD; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphorylation; Principal Investigator; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Protein Isoforms; Protein Phosphorylation; Receptor Cross-Talk; Receptor Protein; Receptor Signaling; Receptors, Opiate; Receptors, Opioid, delta; Receptors, Opioid, mu; Receptors, delta; Receptors, mu; Regulation; Research Personnel; Researchers; Role; Roxanol; SRC; SRC gene; SRC1; Series; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Spinal Ganglia; Statex SR; System; System, LOINC Axis 4; Techniques; Testing; Time; Transcript; Tyr-Ala-Gly-(NMe)Phe-Gly-ol; V (voltage); Vole; ing; abused drugs; analgesia; base; beta-arrestin; beta-arrestin 2; biological signal transduction; c src; c-src Proto-Oncogenes; cellular targeting; desensitization; dorsal root ganglion; drug of abuse; drug/agent; drugs abused; drugs of abuse; field mouse; flow cytophotometry; inhibitor; inhibitor/antagonist; mouse mutant; neuronal; programs; protein expression; receptor; receptor internalization; receptor-mediated signaling; response; social role; trafficking; v-SRC Avian Sarcoma (Schmidt-Ruppin A-2) Viral Oncogene Homolog; voltage
Budget start date: 1-JUN-2009
Budget end date: 31-MAY-2010
5P50DA005010-23_0016 (2009): $251899
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Keywords: Address; Administrator; Advertisements; Advertising; Animal Experimental Use; Animal Experimentation; Animal Housing; Animal Research; Animals; Applications Grants; Award; Book Chapters; Budgets; Carbon Dioxide Snow; Cell Isolation; Cell Segregation; Cell Separation; Cell Separation Technology; Chapters, Book; Charge; Chemicals; Computer Programs; Computer software; Computers; Conflict; Conflict (Psychology); Contracting Opportunities; Contracts; Dry Ice; Ensure; Environment; Equipment; Equipment and supply inventories; Evaluation; Expenditure; Faculty; Fees; Fellowship; Funding; Funding Opportunities; Gases; Grant; Grant Proposals; Grants, Applications; Guidelines; Housing, Animal; Human Resources; Institution; Interview; Inventory; Journals; Laboratories; Lectures; Lectures (PT); Lectures [Publication Type]; Libraries; Licensing; Magazine; Mails; Maintenance; Maintenances; Manpower; Manuscripts; Mediation; Mentors; Microscope; Monitor; Negotiating; Negotiation; Neurosciences; Office Management; Operation; Operative Procedures; Operative Surgical Procedures; Opioid Receptor; PYMT; Participant; Peptide Synthesis; Personnel Staffing; Pilot Projects; Policies; Preparation; Procedures; Process; Program Evaluation; Programs (PT); Programs [Publication Type]; Progress Reports; Publications; Publishing; Radiation; Radioactive; Reagent; Receptors, Opiate; Recommendation; Refuse Disposal; Refuse Disposals; Reports, Progress; Research; Research Resources; Resolution; Resources; Role; SCHED; Safety; Schedule; Schools, Medical; Scientific Publication; Services; Software; Speech; Staffing, Personnel; Structure; Students; Substance abuse problem; Surgical; Surgical Interventions; Surgical Procedure; Training; Training Activity; Training Programs; Travel; Universities; Update; Visit; Work; ing; abuse of substances; abused drugs; animal care; cell sorting; computer network; computer program/software; drug of abuse; drugs abused; drugs of abuse; graduate student; lectures; medical schools; meetings; member; payment; personnel; pilot study; programs; ray (radiation); response; satisfaction; social role; substance abuse; surgery; waste disposal
Budget start date: 1-JUN-2009
Budget end date: 31-MAY-2010
5P50DA005010-23_9010 (2009): $143156
ROLE OF INTERNALIZATION IN THE FUNCTIONAL REGULATION OF OPIOID And ORL-1 RECEPTORS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-150009 from National Institute On Drug Abuse
Sponsored Links Excellgen http://Excellgen.com
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-149004 from National Institute On Drug Abuse
Project start date: 2000-08-05
Project end date: 2001-07-31
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-10 from National Institute On Drug Abuse IRG: SRCD
Project start date: 1987-09-30
Project end date: 1997-09-28
5P50DA005010-10 (1996): $767917
IDENTIFICATION OF RECEPTORS RELATED TO, BUT DISTINCT FROM, DOR-1
Christopher J Evans, Stefan Hatos Professor And Director
Institution:
Grant 5P50DA005010-100005 from National Institute On Drug Abuse
Keywords: molecular cloning, opioid receptor, G protein, complementary DNA, nucleic acid probe, nucleic acid sequence, oligonucleotide, receptor coupling, human genetic material tag, laboratory mouse, laboratory rat, polymerase chain reaction
CORE--ADMINISTRATIVE AND RESEARCH FACILITY
Christopher J Evans, Stefan Hatos Professor And Director
Institution:
Grant 5P50DA005010-109001 from National Institute On Drug Abuse
Keywords: biomedical facility, nucleic acid chemical synthesis, peptide chemical synthesis, biomedical equipment resource, nucleic acid sequence, oligonucleotide, protein sequence, radioactive waste, sanitation, waste disposal, laboratory mouse, laboratory rat, mass spectrometry, tissue /cell culture
IDENTIFICATION OF RECEPTORS RELATED TO, BUT DISTINCT FROM, DOR-1
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-090005 from National Institute On Drug Abuse
CORE--ADMINISTRATIVE AND RESEARCH FACILITY
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 5P50DA005010-099001 from National Institute On Drug Abuse
Grants awarded to Christopher J Evans
ROLE OF INTERNALIZATION IN THE FUNCTIONAL REGULATION OF OPIOID And ORL-1 RECEPTORS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-110009 from National Institute On Drug Abuse
Abstract: To study the trafficking of mu, delta and ORL-1 receptors and their ligands following receptor activation. a). What is the cellular distribution and/or vesicular localization of mu receptors both in vitro and in vivo following various acute and chronic agonist treatments. In vitro, is the trafficking of the delta and ORL-1 receptors similar to that of the mu receptor? b) With the development and use of fluorescent opioid peptide ligands, can we study the trafficking of ligands and receptors be observed in real time. C) What are the kinetics of recover of surface opioid and ORL-1 receptors following agonist treatment both in vitro and in vivo? What are the processes involved in the recovery of surface receptor in vitro. 2. To identify the structural features of opioid ligands which are critical for triggering mu, delta,, and ORL-1 receptor sequestration and determine if any correlation exists between a ligand s ability to trigger internalization and its potency or efficacy for other receptor functions. a) What structural features of peptide and alkaloid ligands are required to induce sequestration of mu, delta and ORL-1 receptors in vitro. b) What is the relationship between a ligand s ability to trigger internalization and its binding affinity potency or efficacy in inhibition of cAMP accumulation and stimulation of GTPgammaS binding in mu, delta and ORL-1 receptors. c) Is an agonist s capacity to cause internalization the same in vivo as in vitro for mu, delta and ORL-1 receptors? 3. To determine the functional significance of sequestration of mu receptors. a. Is there a difference in desensitization or resensitization of the mu receptor in vitro between agonists that induce sequestration verses those that do not? b. Do mu agonists that induce internalization have the same effects on long-term potentiation (LTP) as those which do not? c. How do adaptational processes differ between wild type and internalization-deficient mutant mu receptors? d. What are the behavioral manifestations and pharmacological consequences of opioid treatment in a mouse expressing an internalization-deficient mu receptor? 4. To determine whether mu, delta, or ORL-1 receptor internalization can be observed under conditions which induce the endogenous release of opioid peptides. a. Can receptor sequestration be observed in the CNS following stress- induced analgesia (SIA) using the swim stress model? b. Does amygdala kindling result in receptor sequestration? c. Does stimulation of the perforant pathway of the hippocampus result in receptor sequestration?
Keywords: opioid receptor, protein structure /function, receptor binding, receptor expression, ligand, neuropeptide, opioid, protein transport, autoradiography, fluorescent dye /probe, laboratory mouse, molecular cloning, peptide chemical synthesis, transgenic animal, video microscopy
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-07 from National Institute On Drug Abuse IRG: SRCD
Abstract: This continuation of the NlDA Center at UCLA will further characterize key molecules and mechanisms involved in opioid receptor regulation with the broad goal of more clearly understanding the processes of tolerance, dependence and withdrawal associated with drugs of abuse. This basic research ultimately will permit the development of human probes and reagents for clinical investigations addressing differential diagnosis and treatment. The focus of the continuing research of the Center is based primarily upon our successful isolation of a delta opioid receptor gene (DOR-1) and draws upon the breadth of opioid pharmacological and neurochemical accomplishments of the Center. The program comprises five major research areas 1) The characterization of gene(s) and mRNAs encoding DOR-1. 2) The determination of the structural basis for opiate alkaloid and opioid peptide recognition and for signal transduction. 3) The identification and structural characterization of opioid receptors related to, but distinct from, DOR-1. 4) The analysis of opioid receptor neuroanatomical localization and relationship with other neurotransmitter systems implicated in drug abuse. 5) The understanding of mRNA regulation, post-translational modifications and intracellular trafficking of opioid receptors in both normal and drug perturbed states. The resources and expertise incorporated in the Center span techniques of molecular biology (cDNA cloning, mRNA analysis, genomic analysis, mutagenesis and expression), protein and peptide chemistry (peptide synthesis, peptide and protein purification, protein sequencing), analytical neurochemistry (mass spectroscopy, immunoassay, amino acid analysis), neuropharmacology (receptor binding, second messenger systems, microdialysis, behavior), and histochemistry (immunocytochemistry and in situ hybridization). Functionally the Center s research areas are highly integrated, interactive and interdependent. The continued application of this successful multidisciplinary and collaborative approach will lead to a more precise understanding of the molecular mechanisms underlying the addictive process and thus provide a basis for improved clinical methods for ameliorating the problems associated with drugs of abuse.
Keywords: neurotransmitter, opiate alkaloid, opioid receptor
Project start date: 1987-09-30
Project end date: 1997-07-31
2P50DA005010-07 (1993): $613820
DIFFERENTIAL SIGNALING OF OPIOID DRUGS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-160009 from National Institute On Drug Abuse IRG: ZDA1
Abstract: The variable actions of opioid drugs in vivo are generally attributed to a combination of factors including receptor selectivity and efficacy for signaling, pharmacokinetics, metabolism, and ability to reach target receptors. More recently, opioid drugs have been postulated to display differential selectivity in the activation of opioid receptor homodimers/heterodimers, and in differential triggering of adaptive responses such as internalization. Here we propose that aspects of drug action may be attributed to selective opioid receptor-mediated signaling, and that different ligands, including the plethora of endogenous opioid peptides, may differentially trigger various signaling pathways via the same receptor, a phenomenon we term ligand-directed signaling. This issue will be addressed in Aim l using cell lines transfected with opioid receptors and measurement of pathways predominantly regulated by the alpha or the beta/gamma subunits of G proteins. Our preliminary data suggests that in vitro, inhibition of adenylate cyclase and activation of both MAPK and Akt can be regulated independently by different ligands following opioid receptor activation. Additionally, in mouse brain following acute morphine treatment, Akt activation appears to occur in mu expressing cells whereas MAPK activation occurs in different cells. Thus a related theme of Component II, Aims 2 through 4, will address the in vivo signaling pathways triggered by opioid ligands. We will determine why activation of the Akt pathway, but not the MAPK pathway, is specific to mu-expressing cells and whether the MAPK-activated cells are excitatory or inhibitory neurons. We will also compare the effects of acute and chronic treatment with various opioid agonists and antagonists in selectively activating these kinase pathways. These experiments will be performed in brain and spinal cord from both wildtype mice and those lacking components of the endogenous opioid or ORL-1 systems. This research could ultimately provide some insights into the role of specific signaling pathways on selective behaviors, such as analgesia, reward, sensitization or tolerance exhibited by opioid drugs.
Keywords: biological signal transduction, ligand, neuropharmacology, opiate alkaloid, opioid, opioid receptor, G protein, brain, cAMP response element binding protein, cerebral cortex, drug tolerance, enzyme activity, inhibitor /antagonist, mitogen activated protein kinase, naloxone, phosphorylation, phosphotransferase, receptor sensitivity, spinal cord, stimulant /agonist, cell line, gene targeting, laboratory mouse, transfection, transgenic animal
Project start date: 2002-08-01
Project end date: 2007-07-31
TRANSLATIONAL METHODS/FACILITIES CORE (TMF - CORE) (8 OF 8)
Christopher J Evans, Hatos Professor
University Of California Los Angeles, Office Of Research Administration, Los Angeles, Ca 90095
Grant 5PL1NS062410-04 from Office Of The Director, National Institutes Of Health
Abstract: The Translational Methods/Facilities (TMF) Core will provide state of the art genetic analysis as well as mouse model development, characterization and assessment for all components of the Consortium for Neuropsychiatric Phenomics. Three distinct but interactive units operationally define the specific aims of the TMF-Core ¿ The Genetic Studies Unit will be responsible for generating and analyzing all genetic data in the multiple association studies to be conducted by the Consortium. ¿ The BAG Transgenic Unit will create novel murine models using BAG transgenes or by crossing established mutant lines to a) test the cellular etiology of genotype/phenotype associations by genetic rescue strategies, b) drive cell-specific expression of fluorescent proteins to identify CNS circuitry that may be perturbed via mutant genes and c) model the contribution of candidate genes identified and/or anticipated from ongoing genetic association analysis to significantly modify memory processes or response inhibition. ¿ The Rodent Phenotyping Unit in close collaboration with the Human Translational Applications Core will design, validate and conduct behavioral assays to phenotype mouse models used by the Consortium. The three units leverage considerable physical and intellectual resources at UCLA. Such leveraging will enable the TMF-Core to cost-effectively and efficiently provide informational genetic/phenotype databases, shared animal models and validated behavioral assessments throughout the Consortium and subsequently to the research community outside of UCLA
Keywords: Abnormal coordination; Animal Model; Animal Models and Related Studies; Arts; Assay; Behavior assessment; Behavioral Assay; Bioassay; Biocompatible Materials; Biologic Assays; Biological; Biological Assay; Biomaterials; California; Candidate Disease Gene; Candidate Gene; Causality; Cells; Co-ordination disorder; Collaborations; Communities; Complex; Consultations; Coordination Disorder; Core Facility; DNA; Data; Data Banks; Data Bases; Databank, Electronic; Databanks; Database, Electronic; Databases; Deoxyribonucleic Acid; Development; Dyscoordination; Educational Mainstreaming; Ensure; Etiology; Faculty; Gene variant; Genes; Genetic; Genetic Diversity; Genetic Variation; Genetic analyses; Genome; Genomics; Genotype; Goals; Human; Human, General; Incoordination; International; Investigation; Investigators; Lack of Coordination; Mainstream Education, achievement; Mainstreaming; Mainstreaming (Education); Mammals, Mice; Mammals, Rodents; Man (Taxonomy); Man, Modern; Methods; Mice; Modeling; Murine; Mus; Operation; Operative Procedures; Operative Surgical Procedures; Phenotype; Population; Preparation; Process; Programs (PT); Programs [Publication Type]; Proteins; Research; Research Activity; Research Design; Research Personnel; Research Resources; Researchers; Resources; Rodent; Rodentia; Rodentias; Sampling; Study Type; Surgical; Surgical Interventions; Surgical Procedure; Testing; Time; Transgenes; Transgenic Organisms; Universities; Variation (Genetics); allelic variant; association test; base; behavioral assessment; clinical data repository; clinical data warehouse; cost; data management; data repository; design; designing; disease causation; disease etiology; disease/disorder etiology; disorder etiology; gene product; genetic analysis; genetic association; genome-wide; member; memory process; model development; model organism; mouse model; mutant; neuropsychiatric; neuropsychiatry; novel; phenomics; programs; relational database; response; study design; success; surgery; transgenic; vervet
Project start date: 2007-09-30
Project end date: 2012-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
PFA/PA: RFA-RM-06-008
5PL1NS062410-04 (2010): $653140
5PL1NS062410-03 (2009): $503565
5PL1NS062410-02 (2008): $667744
CORE--TISSUE CULTURE AND NEUROCHEMISTRY
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-169003 from National Institute On Drug Abuse IRG: ZDA1
Abstract: The Tissue Culture and Neurochemistry Core is an essential and critical resource for the full realization of the CSORDA objectives and goals. Nearly every Component of the Center will take advantage of the expertise provided by Core staff scientists and the manifold resources of the Core. The function of the Tissue Culture and Neurochemistry Core is to provide CSORDA members with resources related to the use of primary cell cultures and cell lines, to provide access to the Center s library of established cell lines, and to offer help or advice with transfections, propagation of viral stocks, and establishing cultures of neurons isolated from wild-type or knockout mice. This Core will likewise provide support for experiments which rely on immunohistochemistry, confocal, light, or fluorescent microscopy techniques, as well as provide video imaging and fluorescence activated cell sorting (FACS) expertise. In addition, this Core will provide key analytical neurochemistry support such as assays for intracellular cAMP levels, mitogen-activated protein kinase activity, radiolabeled ligand binding studies, radioimmunoassays (RIA), and ELISA. Finally, the Tissue Culture and Neurochemistry Core will consult with the various Principal Investigators and Core supervisors on overall strategies for individual projects and provide training, education, and support in selected tissue culture and neurochemical methods to students, technicians, visiting scientists, as well as collaborators.
Keywords: biomedical facility, neurochemistry, tissue /cell culture, cyclic AMP, enzyme activity, mitogen activated protein kinase, neuron, opiate alkaloid, opioid, opioid receptor, training, cell line, confocal scanning microscopy, enzyme linked immunosorbent assay, flow cytometry, fluorescence microscopy, gene targeting, immunocytochemistry, laboratory mouse, microorganism culture, radioimmunoassay, radiotracer, transfection, transgenic animal, video microscopy
Project start date: 2002-08-01
Project end date: 2007-07-31
Center For Study Of Opioid Receptors And Drugs Of Abuse
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-16 from National Institute On Drug Abuse IRG: ZDA1
Abstract: The research objectives of CSORDA are to gain insights into the mechanisms of action of opioid drugs at their cognate receptors with the goal of discerning molecular and cellular processes that contribute to opioid-induced behaviors as well as adaptations leading to opioid addiction, tolerance and withdrawal. The Center has six integrated components, which will investigate the activity of opioid ligands at the molecular, cellular and behavioral levels utilizing overlapping methodologies and resources. The Components of the Center will specifically investigate I) Constitutive activity of opioid receptors as a potential target for the development of therapeutic drugs. II) Differential signaling among opioids and targeting of ligands to selective signaling pathways. III) Mechanisms of mu/delta-opioid receptor interactions that influence cellular signaling. IV) Desensitization of opioid signaling in dorsal root ganglia. V) The endogenous, opioid system as a regulator of hedonic homeostasis. VI) The role of memory and fear in opioid adaptive responses. In addition to the research specified within the components, a Pilot Program will be implemented to create new avenues of investigation within the Center and enrich the ongoing programs. For facilitation of the component and pilot programs, the Center has an Administrative Core and three Scientific Cores, which serve to integrate resources, as well as provide practical expertise and training in specific technical areas. The Scientific Cores cover molecular biology, tissue culture, mutant animal breeding/genotyping and various neurochemical procedures. Continued application of this multidisciplinary and collaborative approach will enhance our understanding of the molecular mechanisms underlying opioid actions and provide a basis for improved opioid pharmacotherapies as well as clinical approaches to ameliorating problems associated with drugs of abuse.
Keywords: opiate alkaloid, opioid, opioid receptor, gene targeting, laboratory mouse, transgenic animal
Project start date: 1987-09-30
Project end date: 2007-05-31
2P50DA005010-16 (2002): $1117954
CORE--ADMINISTRATIVE AND PILOT PROGRAM
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-169007 from National Institute On Drug Abuse IRG: ZDA1
Abstract: The Administrative Core provides programmatic structure, logistical support, and mediation of the Center s interactions with the training programs, administrative offices, and scientific and governmental institutions. The Core also is responsible for overseeing the Pilot Program. Day-to-day operation of the Core is the responsibility of the Core Supervisor (Terry Novorr), with immediate oversight by the Administrative Core Director, Dr. Chris Evans or in his absence, the Associate Director Dr. Nigel Maidment. Responsibilities of the Core include Coordination and evaluation of the Center s scientific directions, educational roles, progress and interactions. Overseeing operation of the Pilot Program; including advertising for potential grantees, evaluation of proposals and integration with ongoing research in CSORDA. Resolving personnel issues including hiring, conflicts, promotions and replacements. Logistical support for publications, grant preparations, coordinating of meetings, travel and purchasing. Budgetary preparation, prioritization and monitoring. The Administrative Core is essential to the operation of CSORDA.
Keywords: biomedical facility, health science research, substance abuse, health science research analysis /evaluation, opiate alkaloid, opioid, opioid receptor, postdoctoral investigator
Project start date: 2002-08-01
Project end date: 2007-07-31
Center For Study Of Opioid Receptors And Drugs Of Abuse (CSORDA)
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-21 from National Institute On Drug Abuse IRG: ZDA1
Abstract: Center for study of opioid receptors and drugs of abuse (CSORDA) the research objectives of CSORDA are to gain insights into the mechanisms of action of endogenous opioids and opioid drugs at their cognate receptors with the goal of discerning fundamental processes contributing to behaviors such as analgesia, addiction, tolerance and withdrawal. The Center has four integrated components, which will investigate the activity of opioid ligands at the molecular, cellular and behavioral levels utilizing complementary methodologies and shared resources. The Components of CSORDA will specifically investigate I) The regulation of trafficking and signaling of mu and delta receptors in vivo by high resolution imaging of mice expressing fluorescent receptors; II) The basis for mu ligand- directed signaling and receptor trafficking using primary dorsal root ganglia cells from wild-type as well as receptor and arrestin-deficient mice; III) The role of the endogenous opioid system in goal-directed and habitual behaviors both in drug free and opioid dependent conditions; IV) The functional regulation of transcripts in the striatonigral and striatopallidal projection neuron circuits and their relation to opioid drug reward. In addition to the research specified within the components, a Pilot Program will continue to create new avenues of investigation within the Center and enrich the ongoing programs. For facilitation of the component and pilot projects, an Administrative Core and the CSORDA Mouse Core (CMC), serve to integrate resources and coordinate the needs of the center. The overall mission of CSORDA is to enhance the understanding of the mammalian opioid system thereby providing strategies for improving the clinical use of opioids and treatments for addition.
Project start date: 1987-09-30
Project end date: 2012-05-31
2P50DA005010-21 (2007): $1160461
Sponsored Links Excellgen http://Excellgen.com
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 3P50DA005010-15S1 from National Institute On Drug Abuse IRG: ZDA1
Abstract: Applicant s ) The research objectives of the Center for Study of Opioid Receptors and Drugs of Abuse are to gain insights into the mechanisms of action of opioid drugs and their receptors with the ultimate goal of discerning molecular processes that contribute to opiate addiction, tolerance and withdrawal. The Center has four integrated components which have evolved since the cloning of the opioid and ORL-1 receptors. These components will investigate the actions and regulation of receptors both in vitro and in vivo utilizing overlapping methodologies and resources. The Center will specifically investigate 1) The coupling of opioid and ORL-1 receptors through G-proteins and second messenger systems to the release process, with an emphasis on the contribution of specific calcium channel subtypes. 2) The role of receptor internalization in the regulation of receptors and the functional consequences of drugs that induce sequestration (i.e. enkephalins) versus those that do not (i.e. morphine). 3) The role of ORL-1 receptor ligands and their interaction with opiate drugs in the modulation of hippocampal function. 4) Whether ORL-1 receptor agonists have "anti-opioid" effects on brain reward systems. To facilitate the research, the Center has an Administrative Core and five Scientific Cores which serve to integrate its resources, as well as provide practical expertise in specific technical areas. The Scientific Cores cover Molecular Biology, Peptide Chemistry, Analytical Neurochemistry, Tissue Culture and Imaging of both antibody and nucleic acid probes. The continued application of this multidisciplinary and collaborative approach is enhancing our understanding of the molecular mechanisms underlying the addiction process and thus provides a basis for improved clinical methods for ameliorating the problems associated with drugs of abuse.
Keywords: neuropeptide, opiate alkaloid, opioid, opioid receptor
Project start date: 1987-09-30
Project end date: 2002-09-29
3P50DA005010-15S1 (2002): $268142
3P50DA005010-13S1 (1999): $54178
2P50DA005010-11 (1997): $1041761
MUTATIONAL LIBRARIES TO STUDY OPIOID RECEPTOR FUNCTION
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 1R03DA011962-01 from National Institute On Drug Abuse IRG: ZDA1
Abstract: Applicant s ) Point mutations have proven to be powerful for the identification of structural amino acids involved in Gprotein coupled receptor function. However, the generation of appropriate mutants necessitates a preconceived hypothesis regarding which amino acids may be relevant. For a number of receptor functions it is difficult to assume a priori the involvement of particular amino acids and what properties of the amino acids might be important. This proposal aims to generate region-selective random mutational libraries of the mu-opioid receptor using PCR strategies. Initially these libraries will be used to screen for mutations that perturb mu-opioid receptor sequestration. Previously we have found that endogenous opioid peptides and a subset of alkaloids can induce sequestration of the mu-opioid receptor, whereas morphine and many clinically-used opiate drugs do not trigger receptor internalization, or do so only weakly. This suggests that agonist ligands which have similar effects on receptor-mediated signaling can have dramatically different effects on receptor trafficking. The aim of the study is to use the random mutational libraries to understand the molecular basis for ligand-dependent receptor internalization. In the future, the results should help to clarify the significance of internalization in the development of opiate drug-induced tolerance and dependence.
Keywords: morphine, opioid receptor, protein structure /function, receptor binding, receptor mediated endocytosis, complementary DNA, intracellular transport, nucleic acid sequence, protein transport, recombinant protein, flow cytometry, peptide library, polymerase chain reaction, site directed mutagenesis, tissue /cell culture
Project start date: 1998-07-01
Project end date: 2000-05-31
1R03DA011962-01 (1998): $75833
Translational Methods/Facilities Core
Christopher J Evans, Stefan Hatos Professor & Director
University Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 1PL1NS062410-01 from National Institute Of Neurological Disorders And Stroke IRG: ZRR1
Keywords: behavioral genetics, biomedical facility, neuropsychology, phenotype NIH Roadmap Initiative tag, clinical research, human genetic material tag, laboratory mouse
Project start date: 2007-09-30
Project end date: 2012-06-30
1PL1NS062410-01 (2007): $975318
ROLE OF INTERNALIZATION IN THE FUNCTIONAL REGULATION OF OPIOID And ORL-1 RECEPTORS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 3P50DA005010-15S10009 from National Institute On Drug Abuse
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 3P50DA005010-15S19004 from National Institute On Drug Abuse
Project start date: 2001-08-01
Project end date: 2002-09-29
ROLE OF INTERNALIZATION IN THE FUNCTIONAL REGULATION OF OPIOID And ORL-1 RECEPTORS
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 3P50DA005010-13S10009 from National Institute On Drug Abuse
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 3P50DA005010-13S19004 from National Institute On Drug Abuse
Project start date: 1999-08-01
Project end date: 2000-07-31
CENTER FOR STUDY OF OPIOID RECEPTORS AND DRUGS OF ABUSE
Christopher J Evans, Stefan Hatos Professor & Director
University Of California Los Angeles
office Of Research Administration
los Angeles, Ca 90095
Grant 3P50DA005010-10S1 from National Institute On Drug Abuse IRG: SRCD
Project start date: 1987-09-30
Project end date: 1997-09-28
3P50DA005010-10S1 (1997): $130352
Sponsored Links Excellgen http://Excellgen.com
Christopher J Evans, Stefan Hatos Professor And Director
University Of California Los Angeles Office Of Research Administration Los Angeles, Ca 90095
Grant 2P50DA005010-119004 from National Institute On Drug Abuse
Keywords: biomedical facility, peptide chemical synthesis, protein engineering, protein purification, antigen, opioid receptor, synthetic peptide
Project start date: 1997-09-30
Project end date: 1998-07-31