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center for substance abuse research

Research Programs

 

The objective of the Center for Substance Abuse Research (CSAR) is to provide the core facilities and the intellectual milieu to support research of funded investigators devoted to exploring how drugs of abuse interact with a variety of systems of the body to alter function.  Approaches include research projects devoted to understanding the receptors for these drugs, their active sites, how they are triggered, and their signal transduction pathways.  For example, we are investigating receptor desensitization and heterodimer formation as it alters function at the biochemical, cellular, tissue, and whole-animal levels.  These studies have put researchers in CSAR at the forefront in delineating the pharmacologic mechanisms and physiological significance of pathways within the nervous system, as well as pathways connecting the nervous and immune systems.  The studies bear on consequences of interaction of drugs of abuse and related endogenous ligands with their receptors as they modulate immune responses, body temperature regulation, and pain perception.  In addition, a major focus of studies utilizing the P30 Center has been drug interactions.  It is well established that addicts rarely abuse only a single drug.  We are investigating basic aspects of drug interactions for effects on signal transduction at the cellular level, as well as physiological processes in the whole animal.  These experimental studies are built on continuing expansion of theoretical advances in understanding the pharmacology of drug interactions carried out by one of our members.  This type of approach, supported by the Cores of the P30 Center Grant, differs from that taken by P30 Centers whose focus is on prevention and treatment.  We believe that exploration of the basic pathways utilized by drugs of abuse will lay the foundation for understanding the biological basis of drug addiction, for development of future therapies, and for providing the scientific basis for new approaches to prevention and treatment. We make the analogy to cancer research, where understanding of the biological basis for neoplasia has led to new fundamental concepts in the causes of cancer and how it spreads, with concomitant new approaches to treatment and prevention.  It is our belief that pursuit of the biology of opioids, cannabinoids, cocaine, methamphetamines, and nicotine, alone and in combination, will lead to similar advances in prevention and treatment of drug abuse.

 

The research Cores of the NIDA P30 Center are listed below:

Animal Core

Biochemical Pharmacology Core

Cell and Immunology Core

Integrative Pharmacology Core

Molecular Core

 

 

Animal Core

 

Ellen M. Unterwald, Director

                                                                                                                                           

The Animal Core provides the following services to CSAR:

 

  • Breeding and maintenance of animals that have targeted gene deletions or express transgenes 
  • Genotyping of the animals carried out in collaboration with the Molecular Core
  • Phenotyping of the animals characterized together with the Integrative Pharmacology Core

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Biochemical Pharmacology Core

 

Lee-Yuan Liu-Chen, Director

                                                                                                                             

The Biochemical Pharmacology Core provides the following services:

  • Opioid receptor binding assays and receptor autoradiography and immunohistochemistry for hemagglutinin peptide (HA) or FLAG-tagged opioid receptors
  • Detection of mRNA of opioid receptors and opioid peptides by in situ hybridization
  • Assessment of receptor activation by determination of cAMP levels, [35S]GTPgS binding, and MAP kinase phosphorylation
  • Protein gel electrophoresis and Western blots
  • Limited assessment of levels of drugs following in vivo administration to animals (service now transferred to Analytical Core)

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Cell and Immunology Core

 

Toby K. Eisenstein, Director

                                                                               

The Cell and Immunology Core provides the following services:

 

  • Maintains, prepares, and stores human and murine cell lines and clones
  • Carries out fusions and screening to identify hybridomas with reactivity to opioid receptors; grows and purifies monoclonal antibodies
  • Characterizes cells by their surface markers using Flow Cytometry (FACS)
  • Separates cells by magnetic separation
  • Supports the confocal microscopy laboratory
  • Carries out ELISA and Cytometic Bead Array assays for neuropeptides and cytokines
  • Carries out basic tests of immune function such as response of     lymphocytes to mitogens and natural killer assays
  • Performs mycoplasma testing of cell lines

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Integrative Pharmacology Core

 

Alan Cowan and Ronald J. Tallarida, Co-Directors

 

The Integrative Pharmacology Core provides the following services:

  • Quantitation of overt behavior (mice and rats)
  • Assessment of analgesia (using several different nociceptive stimuli), tolerance, and physical dependence (mice and rats)
  • Evaluation of stomach emptying, small intestinal and colonic transit (mice)
  • Assessment of body temperature effects of different abused drugs
  • Administration of drugs into various brain areas
  • Collection of effluents from various brain areas using microdialysis in non-anesthetized rats

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Molecular Core

 

Thomas J. Rogers, Director

 

In the P30 application funded four years ago, it was proposed that the following services be offered by the Molecular Core:

 

  • General molecular services including:

a.  Vector design and construction

b.  Plasmid preparation for DNA or RNA sequencing

c.  Expression plasmid preparation and cell transformations

d.  Antisense oligonucleotide design

 

  • Gene expression services including:

a.  RNase protection analyses

b.  Quantitative and semi-quantitative RT-PCR design and internal

     standard preparation

c.  Preparation of samples for gene microarrays

 

  • Animal genotyping services

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