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Full text of "Annual report : National Institute on Drug Abuse"

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ANNUAL REPORT 
OF THE 



Addiction Research Center 



National Institute on Drug Abuse 

P.O. Box 5180 
Baltimore, Maryland 21224 



NATIONAL INSTITUTES OF HEALTH 
NIH LlS&ARY 



JAN 3 2008 



BLDG 10, 1 n CENTER DR 
BETHESDA. MD 20892-1150 



U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 

Public Health Service 
Alcohol, Drug Abuse and Mental Health Administration 



Table of Contents 
Director's Overview 
Neurosciences Branch 
Overview 

Molecular Pharmacology Laboratory 
Overview 

Summary of Ongoing Projects 
Publications 
Project Notification Forms 



Z01 DA 001 12-04 MPL 
M.J. Kuhar 

Z01 DA 00107-05 MPL 
M.J. Kuhar 

Z01 DA 0108-04 MPL 
M.J. Kuhar 

Neuropharmacology Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notificaiton Forms 

Z01 DA 00200-05 NPL 
E.D. London 



Z0I DA 00202-07 NPL 
E.D. London 

Z01 DA 00312-01 NPL 
E.D. London 

Z01 DA 00206-06 NPL 
Tsung-Ping Su 

Z01 DA 00210-05 NPL 
Alane Kimes 



Drug Receptors. Neurotransmitters and 
Addiction 

Drug Receptors In Vivo 
The Cocaine Receptor 



Cerebral Effects of Abused Drugs: 
Brain imaging in Humans and Laboratory 

Animals 

Physiological Effects of Opioids 

Ligand-Gated Ion Channels 

Sigma Receptors 

HIV Infection and Drug Abuse 



1 

2 
3 



10 

12 
12 
14 



18 



20 



22 



24 



26 



Neurobiology Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notification Forms 

Z01 DA 00302-03 NBL 
Errol De Souza 

Z01 DA 00303-03 NBL 
Errol De Souza 

Z0I DA 00304-03 NBL 
Errol De Souza 

Z01 DA 00305-03 NBL 
Errol De Souza 

Z01 DA 0031 1-02 NBL 
Errol De Souza 

Z01 DA 00307-03 NBL 
Stan Yeh 

Z01 DA 00308-03 NBL 
Errol De Souza 

Z01 DA 00309-03 NBL 
Errol De Souza 



Neurotoxic Effects of MDA and MDMA 



CRF in Addictive, Neuropsychiatric and 
Neurodegenerative Disorders 

CRF as a Stress Neurotransmitter in the 
Brain-Endocrine-Immune Axis 

Neurotoxicity of Selected Drugs to 
Monoamine Neurons in Brain 

Sigma and PCP Receptors in Neuropsychiatric 
Disorders 

Effects of Cocaine on Monoamines and Their 
Metabolites in Rat Brain 

Role of Sigma Receptors in Endocrine Organs 
and Immune Tissue 

Interlcukjn- 1 in the Brain- Endocrine- 
Immune axis 



28 
27 
28 



34 



36 



38 



41 



43 



45 



47 



49 



Molecular Neurobiology Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notification Forms 

Z01 DA 001 14-02 MNL 
George Uhl 

Z01 DA 001 15-03 MNL 
George Uhl 

Z01 DA 001 16-03 MNL 
Geroge Uhl 



Receptor cDNA Epression Coning Using 
Ligand Autoradiographic Screening 

Receptor cDNA Expression Cloning Using 
Xenopus Oocyte Expression 

Genes Related to Drug Abuse I: Regulation 
of Opioid Peptide Genes 



51 
51 
53 



56 



58 



60 



Z01 DA 001 17-02 MNL 
George Utal 

Z01 DA 00314-01 MNL 
George Uhl 



Genes Related to Drag Abuse II: Central 
Brain Pathways of Reinforcement/Reward 

D2 Receptor Gene Allelic Association with 
Substance Abuse 



63 



65 



Prelinical Pharmacology Branch 
Overview 
Behavioral Pharmacology and Genetic Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notification Forms 



Z01 DA 00001-06 BPL 
Steven Goldberg 

Z01 DA 00003-06 BPL 
Steven Goldberg 

Z01 DA 00009-04 BPL 
Charles Schindler 

Z01 DA 00012-02 BPL 
Rodney Marley 

Z01 DA 00101-05 BGL 
Gregory Elmer 

Z01 DA 00102-05 BGL 
Gregory Elmer 

Psychobiology Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notification Forms 

Z01 DA 00103-01 PBL 
Jonathan Katz 



Control of Behavior by Drug Injection 



Effects of Drugs on Schedule-Controlled 
Behavior in Experimental Animals 

Cardiovascular Changes Induced by Cocaine 



Genetic Factors in Response to Chronic Drug 
Treatment 

Pharmacogenetics: Acute Responses to Drug 
Administration 

Pharmacogenetic Factors in Drug Reinforced 
Behavior 



67 

67 
68 
76 



82 
85 
88 
91 
93 
96 

98 
98 
101 



Basic Mechanisms of Cocaine Effects 



105 



Z01 DA 00104-01 PBL 
Jonathan Katz 

Z01 DA 00105-01 PBL 
Jonathan Katz 



Drug Development 



Behavioral Pharmacology of 
Dopamine Systems 



108 



110 



Clinical Pharmacology Branch 

Overview 

Biology of Dependence and Abuse Liability Laboratory 

Overview 

Summary of Ongoing Projects 

Publications 

Project Notification Forms 



Z01 DA 00037-01 BDL 
Jack Henningfield 

Z01 DA 00028-02 BDL 
Jack Henningfield 

Z01 DA 00029-02 BDL 
Jack Henningfield 

Z01 DA 00033-02 BDL 
Jack Henningfield 

Z01 DA 00038-01 BDL 
Wallace Pickworth 

Z01 DA 00039-01 BDL 
Wallace Pickworth 



Z01 DA 00013-05 BDL 
Charles Haertzen 

Z01 DA 00040-01 BDL 
Jack Henningfield 

Z01 DA 00041-01 BDL 
Jack Henningfield 

Z01 DA 00042-01 BDL 
Jack Henningfield 



Physiologic, Cognitive and Subjective Effects 
of Commonly Abused Drugs 

Assessment of Mazindol for Abuse Liability 



112 

113 
113 
114 



120 



121 



Interaction Between Ethanol and Prostaglandin 
Synthetase Inhibitors 

Nicotine Patch: Effects on Smoking Subjective 
and Physiologic Functions 

Transcranial Electrostimulation Therapy (TCET) 
During Smoking Cessation 

Physiologic and Performance Effects of Ethanol 
and Pentobarbital and Interactions with 
Indomethacin 

Archival Data Base Project 

Opioid Agonist and Antagonist Sensitivity in Opiate 
Experienced Individuals 1 28 

Discriminitive Stimulus Effects of Stimulant Drugs 

129 

Effect of Reinforcement Contingencies on Human 
Task Performance 130 



122 



123 



124 



125 



126 



Z01 DA 00034-02 BDL 
Jack Henningfield 

Z01 DA 00026-02 BDL 
Jack Henningfield 

Z01 DA 00044-01 BDL 
Jack Henningfield 

Z01 DA 00031-02 BDL 
Jack Henningfield 

Z01 DA 00045-01 BDL 
Jack Henningfield 

Z01 DA 00046-01 BDL 
Jack Henningfield 

Z01 DA 00047-01 BDL 
Jack Henningfield 

Z01 DA 00006-04 BDL 
Jack Henningfield 

Z01 DA 00009-06 BDL 
Jack Henningfield 

Z01 DA 00010-06 BDL 
Jack Henningfield 

Z01 DA 00024-03 BDL 
Jack Henningfield 

Z01 DA 00007-05 BDL 
Jack Henningfield 

Z01 DA 00030-02 BDL 
Jack Henningfield 

Z01 DA 00004-05 BDL 
Jack Henningfield 

Z01 DA 00025-03 BDL 
Jack Henningfield 

Z01 DA 00014-03 BDL 
Jack Henningfield 

Z01 DA 00012-07 BDL 
Jack Henningfield 

Z01 DA 00005-05 BDL 
Jack Henningfield 



Opioid Self-Administration in Humans 



Assessment of Opioid Agonists and Antagonists 



Psychotropic Properties of Stimulants and 
Sedatives: Discriminative Properties 

Effects of Nicotine in Nonsmokers 



Lobeline and Nicotine: Subjective, Physiologic 
and Kinetic Effects 

Arterial Kinetics of Smoked Cocaine 



Outpatient Drug Discrimination 

Triazolam Self-Administration: Effects of 
Yohimbine Pretreatment 

Effects of Drugs on Cigarette Smoking and 
Responses to Nicotine 

Behavioral and Pharmacologic Factors in Nicotine 
Replacement for Tobacco Dependence 

Opioid Self-Administration: Humans Compared 
to Animals 

Effects of Commonly Used Drugs on Behavioral 
Performance in Normal Subjects 

Passive Tobacco Smoke: Nicotine Absorption, 
Subjective Effects and Performance 

Comparative Self-Administration (Monkeys and 
Human): Nicotine and Cocaine 

Acquisition of Dependence to Cigarettes and 
Smokeless Tobacco 

Cholinergic Agonists and Antagonists 



Factors Influencing Behavioral and Physiologic 
Response to Opioids (mu Project) 

Abuse Liability of Smokeless Tobacco Products 



131 



132 



134 



136 



138 



139 



140 



141 



142 



144 



146 



148 



149 



150 



152 



154 



155 



157 



Z01 DA 00032-02 BDL 
George Uhl 

Z01 DA 00035-02 BDL 
Jack Henningfield 

Z01 DA 0001 1-05 BDL 
Jack Henningfield 

Z01 DA 00008-04 BDL 
Jack Henningfield 



Dopaminergic Lessions and Subjective 
Effects of Methylphenidate 

Why Do Substance Abusers Seek Help? 
What are Their Worries About that Help? 

Physiologic Dependence to Tobacco: Cigarette 
Withdrawal and Nicotine Substitution 

Behavioral Performance and Physiologic Effects 
of Drugs: Atropine and Diazepam 



Chemistry and Drug Metabolism Laboratory 

Overview 

Summary of Ongoing Projects 

Project Notification Forms 



Z01 DA 00002-CDM 
Edward Cone 

Z01 DA 00003-05 CDM 
Edward Cone 

Z01 DA 00006-04 CDM 
Edward Cone 

Z01 DA 00007-03 CDM 
Edward Cone 

Z01 DA 00008-02 CDM 
Edward Cone 

Z01 DA 00009-02 CDM 
Marylin Huestis 

Z01 DA 00010-02 CDM 
Edward Cone 



Validity Studies of Commercial Drug 
Screening Assays 

Detection of Drugs of Abuse in Human 
Saliva 

Pharmacokinetics and Pharmacodynamics 
of Opiate Analgesics 

Pharmacokinetics and Pharmacodynamics of 
Drugs of Abuse in Hair 

Pharmacokinetics and Pharmacodynamics of 
Methampnetamine 

Fast Action Dynamics of Marijuana 



Methodological Asessment of the Risk of 
Passive Inhalation of Drugs of Abuse 



Neuroendocrinology/Immunology Laboratory 
Overview 

Summary of Ongoing Projects 
Publications 
Project Notification Forms 



159 
160 
161 
162 

164 
164 



166 
168 
170 

172 
174 
175 
177 

179 
179 
180 



Z01 DA 00004-04 NEI 
Elizabeth Dax 



Inhalable Nitrites - Immune Function 
and Abuse Potential 



184 



Z01 DA 00005-04 NEI 
Elizabeth Dax 

Z01 DA 00006-04 NEI 
Elizabeth Dax 

Z01 DA 00007-04 NEI 
Elizabeth Dax 

Z01 DA 00008-04 NEI 
Elizabeth Dax 

Z01 DA 00103-03 NEI 
Elizabeth Dax 

Z01 DA 00011-03 NEI 
Elizabeth Dax 

Z01 DA 00012-03 NEI 
William Lange 

Z01 DA 00013-03 NEI 
Elizabeth Dax 

Z01 DA 00014-02 NEI 
Nancy Pilotte 

Z01 DA 00015-02 NEI 
Nancy Pilotte 

Z01 DA 00016-02 NEI 
Nancv Pilotte 



Z01 DA 00017-02 NEI 
Elizabeth Dax 



Z01 DA 00018-02 NEI 
Elizabeth Dax 



HTV Prevalence: in Depth Survey of 

Baltimore 186 

Cannabinoids and Their Effects on the 

Immune System and Cognitive Function 187 

Neuroendocrine Secretion During Cocaine 

Withdrawal 189 

The Effects of Cocaine on Hormone Secretion 

From the Anterior Pituitary 191 

Development of Monoclonal Antibodies to 

Drugs and Hormones 193 

Neuroendocrine Correlates of HIV Infection 

and the Development of ARC and AIDS 1 94 

HTV Sero-status in Missionaries from Africa 
1968-1983 195 

Neuroendocrine Correlates of Aggressive/ 

Impulsive Behavior 197 

The Effects of Cocaine on Prolactin Secretion from 
Single Cells of the Anterior Pituitary Gland 199 

The Effects of Cocaine on Dopamine Release from 
Hypothalamic Neurons 201 

Effects of Cocaine and Withdrawal from Cocaine 

on Central Receptors for Peptides, Catecholamines 

and Catecholamine Uptake Markers 202 

Cardia Effects of IV Cocaine Administration as 
Measured by Radionucleotide Scanning, 
Echocardiography and Holier Monitoring 204 

The Effects of Chronically Administered Meta- 
chlorophenylpiperazine on Rat Brain Receptors, 
Neurotransmitters and Neuroendocrine Secretion 205 



Etiology Branch 
Overview 
Vulnerability Laboratory 

Overview 



206 



206 



Summary of Ongoing Research 

Publications 

Project Notification Forms 

Z01 DA 00500-01 VL 
David Newlin 

Z01 DA 10701-02 VL 
David Newlin 



Z01 DA 00501-01 VL 
David Newlin 

Z01 DA 00502-01 VL 
David Newlin 

Z01 DA 00504-01 VL 
David Newlin 



Individual Differences in Cocaine Craving: 
Physiological and Affective Correlates 

Alcohol Ingestion Increases Self-Report of 
Cocaine-Craving: Individual Differences 
in Craving 



206 
208 



210 



212 



Intravenous Morphine Produces Initial Heart 

Rate Increases and Cardiac Vagal Tone Decreases 214 

Cardiovascular Responses to Naloxone-Precipitated 
Withdrawal 216 

Acute Intravenous Cocaine Reduces Cardiac Vagal 
Tone in Cocaine Abusers 218 



Z01 DA 00505-01 VL 
David Newlin 



Z01 DA 00506-01 
David Newlin 



Z01 DA 09601-03 VL 
Geroge Uhl 

Z01 DA 00507-01 VL 
David Newlin 



Z01 DA 06801-04 VL 
Ronald Heming 

Z01 DA 05801-04 VL 
Ronald Heming 

Z01 DA 02101-05 VL 
Ronald Heming 

Z01 DA 02001-05 VL 
Ronald Heming 

Z01 DA 03101-05 VL 
Ronal Heming 



The ARC Drug Expectancy Questionnaire: An 
Instrument for Assessing Expectations Concerning 
use of Cocaine, Heroin. Marijana, Alcohol, and 
Nicotine 

Acute Effects of Various Abused Drugs on Heart 
Rate and Cardiac Vagal Tone: a Common Factors 
Approach 

Antagonists- Withdrawal Up-Regulation of 
of Endogenous Opiate 

Vagal Tone and Attention in 8 to 1 2 Year-Old 
Males Exposed to Opiates in Utero: a 
Preliminary Report 

Cognitive Neurophysiologic Signs of 
Cocaine Abstinence 

Mapping the Effects of Cocaine by EEG 



Acute Abstinenence from Tobacco: Electro- 
Physiological and Cognitive Signs 

Mapping the Effects of Opioid Agonists by EEG 



Effects of Atropine on Cognitive 
Information Processing 



220 



222 



224 



226 



228 



230 



232 



234 



236 



Z01 DA 00508-01 VL 
Ronald Heming 



Neurocognitive Status of Young Boys 
Exposed to Opiates in Utero 



238 



Treatment Branch 

Overview 

Pharmacotherapy Laboratory 

Overview 

Summary of Ongoing Research 

Publications 

Project Notification Forms 

Z01 DA 00139-01 PTL 
David Gorelick 

Z01 DA 00100-01 PTL 
Lawrence Cheskin 

Z01 DA 00101-01 PTL 
David Gorelick 

Z01 DA 00107-01 PTL 
David Gorelick 

Clinical Trials Laboratory 

Overview 

Summary of Ongoing Research 

Project Notification Forms 

Z01 DA 00090-02 CTL 
Rolley Johnson 

Z01 DA 00081-03 CTL 
Lino Covi 



Z01 DA 00091-01 CTL 
Lawrence Cheskin 

Z01 DA 00508-01 CTL 
Lino Covi 

Z01 DA 00098-02 CTL 
Lino Covi 



Effects of Carbarn azepine on Cocaine 
Self- Administration 

Comparison of Detoxification Treatment 
of Opiate Addiction 

HIV Infection. High-Risk Behaviors, and 
Drug Abuse Treatment 

Esterase Activity in Human Cocaine Abusers 



Buprenorphine/Methadone Comparison - 
Maintenance and Detoxification 

Double-Blind Comparison of Desipramine, 
Fluoxetine and Bromocriptine for the Treatment 
of Cocaine and PCP Abuse 

Assessment of Nalmefene Glucuronide as 
a Selective Antagonist of Gut Opioid Action 

Reasons for seeking drug abuse treatment 



Impact of Differing Intensities of Drug Abuse 
Counseling 



239 

239 
240 

241 



245 
246 
247 
249 

250 
250 



252 



253 



254 



256 



258 



ZO 1 DA 00 1 06-0 1 CTL Evaluation of Methadone Maintenance 

Jonn Ball Treatment for Opiate Dependence 260 



Annual Report of the 

Addiction Research Center 

National Institute on Drug Abuse 

January 1, 1990 - December 31, 1990 

Roy W. Pickens, Ph.D., Director 



In 1990. scientists at the Addiction Research Center (ARC) made major contributions across a broad 
range of drug abuse issues. These contributions significantly increased our understanding of the causes, 
consequences, and treatments of drug abuse. They included further progress in characterizing the 
dopamine transporter, which has been shown to be strongly associated with the reinforcing properties of 
cocaine; further studies of the sigma receptor and its role in drug abuse; clarification of the sites in the 
brain at which hallucinogens act; progress toward developing possible medications for treating cocaine 
overdoses; studies of genetic factors associated with opiate and cocaine abuse which may eventually be 
important in the development of prevention and treatment programs; studies of the safety of 
carbamazepine, a drug which may be useful in the treatment of cocaine abuse; studies of the mechanisms 
involved in the cardiovascular effects of cocaine: clinical studies of nalmefene and buprenorphine, two 
drugs which may be useful in the treatment of opiate addiction; and a clinical study of carbamazepine 
for the treatment of cocaine abusers. These and other studies are described in this Annual Report. 

In addition to the strides made in the ARCs science program mentioned above and described in detail in 
the following pages, a number of management initiatives were undertaken in 1990 to improve the 
oversight of the science program and to build a foundation for a stronger program in the future. In 
September, 1990. the ARC held the first of what will become annual Laboratory Reviews. These reviews 
are designed to provide the Director of the ARC with a formal presentation of the work of individual 
laboratories over the past year. A major effort was initiated in 1990 to provide for future expansion of 
several of the ARCs programs by obtaining new space. Approximately 1 2,000 square feet of remodeled 
space has been obtained in the "G Building," a building adjoining the ARCs building on the east side. 
This building will be used to expand our clinical ward from 18 to 32 beds, to provide space for an 
expanded PET program, and to expand our out-patient treatment research program from 40 to 80 slots. 
The out-patient treatment program will be operated under contract to provide basic drug abuse treatment 
services, allowing ARC staff to focus on innovative treatment research studies. The creation of this out- 
patient facility will also allow the ARC to expand its present Etiology program by providing it with the 
space now being used for treatment out-patient studies. The ARC has also obtained approximately 9,000 
square feet of new space in the Triad Building, a building located about a quarter of a mde from the ARC 
building. This space will be used primarily to expand our animal research program which is facing 
severe space shortages in our current building. 

Training is an integral part of the ARCs research program. Training programs at the ARC range from 
formal post-doctoral research fellowships to special summer training programs for high school science 
teachers. In 1990 the ARC provided training to 29 staff fellows, 7 visiting scientists, 17 visiting fellows, 
2 IP As and 1 IRTA. Training was provided to scientists from Japan, China, Hungary, Israel, Spain, the 
United Kingdom, Belgium, Denmark. Columbia, Italy, Poland, Argentina, India and Australia, as well as 
to scientists from the United States. 

The Minority Recruitment and Training Program (MRTP) was established in 1990. The general goal is 
to increase the participation of minorities in research and training across the broad spectrum of activities 
at the ARC. Specifically, this involves increasing the number of minority persons who are trained for 
careers in drug abuse research, increasing the participation of minority persons in the design and 
execution of ARC research, increasing the sophistication of ARC scientists in the design of research 
relevant to minority concerns, and to enhance the access of minority researchers to ARC physical and 
human resources. Progress was made in each of these areas by efforts which included the following: 
visits by ARC staff to cluster groups of Historically Black Colleges and Universities throughout the 



country to establish collaborative working relationships and obtain advice; development of contracts with 
minority professionals to assist in the establishment of training opportunities for minority persons at the 
Addiction Research Center, collaborative efforts with Historically Black Colleges and Universities in the 
Baltimore/Washington area in the development of conferences and training programs for undergraduate 
students: recruitment of minority scientists who are assisting in development of research via our guest 
worker and guest speaker programs, and consultantships. 

In 1990, the Student Fellowship Program was supported primarily by a grant from Marion Merrell Dow 
Inc. for the fifth consecutive year. A total of 17 students, representing 11 colleges or universities, 
participated in the summer program and were placed in 12 laboratories throughout the ARC. On the 
basis of their interests, students were assigned to work directly with an ARC scientist on a specific 
research project. Additionally, students were encouraged to attend seminars, journal clubs, and clinical 
rounds to enhance their knowledge of drug abuse and to broaden their educational experience at the 
ARC. 



Neurosaence Branch 

Michael J. Kuhar, Ph.D. ■ Chief 

Introduction 

The Neurosaence Branch carries out interdisciplinary research to elucidate the mechanisms of action and 
the effects of abused drugs on biological systems. A major goal is to develop new knowledge which can 
be used in the development of new treatment and prevention strategies and new medications for drug 
abusers. Research areas include drug and neurotransmitter receptors, brain imaging, molecular biology, 
and the neuroanatomy and neurochemistry of reinforcement. 

The Neuroscience Branch is composed of four laboratories: The Molecular Pharmacology Laboratory 
directed by Dr. Michael J. Kuhar, the Neuropharmacology Laboratory by Dr. Edythe London; and the 
Molecular Neurobiology Laboratory by Dr. George Uhl. The Neurobiology Laboratory was formerly 
directed by Dr. Errol De Souza. who left the ARC this past year. A search committee is currently 
evaluating applications to fill this position. 

The Branch has been productive in a variety of areas. New chemical probes for the cocaine 
receptor/dopamine transporter have been characterized in in vitro and in vivo binding studies with the 
ultimate goal of characterizing the transporter and in carrying out brain imaging studies. PET scan 
studies of human volunteers have demonsi rated a fundamental role of the amygdala in cocaine induced 
euphoria. A new, useful approach to study amphetamine uptake into nerve terminals has been developed. 
Receptor genes have been studied: a novel GABA receptor cDNA has been cloned and linkage between 
the human dopamine D2 receptor gene and substance use was excluded. 

The research of the Neuroscience Branch has received national and international recognition and 
attention. Dr. Michael Kuhar received the A. Ross Mclntyre Award, which is given by the University of 
Nebraska Medical Center for an outstanding research achievement. He was also invited to give the first 
Ivan Davidson Memorial Lecture at the Bowman Gray School of Medicine. Dr. George Uhl received the 
NIDA Director's Award for outstanding achievement in establishing molecular biological and clinical 
programs. Dr. Edythe London received numerous invitations to present findings at national and 
international meetings. Dr. Errol De Souza received the Jordi Foch-Pi Award from the American Society 
for Neurochemistry for work carried out at the ARC. Dr. Arthur Weissman, a staff fellow in the 
Neurobiology laboratory, was awarded the ARC Staff Fellow Research Award. 



1. Molecular Pharmacology Laboratory - Michael J. Kuhar, Ph.D., Chief 

Overview 

The focus of the Molecular Pharmacology Laboratory is the mesolimbic dopaminergic system and its 
relationship to drugs of abuse. Because of the strong evidence that mesolimbic dopaminergic neurons 
may be a final common pathway for some drugs of abuse, and because of the strong evidence that the 
psychostimulants, particularly cocaine, have receptors at the dopaminergic nerve terminal, we are 
currendy focusing on elements of the dopaminergic synapse. A particular focus is the dopamine 
transporter, the "receptor" for cocaine. Current studies of the transporter include: a detailed structure- 
activity investigation using nearly a hundred analogues of cocaine and mazindol; efforts to purify and 
characterize the transporter protein: development of new ligands for both binding studies and PET and 
SPECT imaging; and behavioral testing of new cocaine analogues. 



Summary of Ongoing Research: 

A. The Cocaine Receptor. 

Because of the identification of the dopamine transporter as the cocaine receptor related to drug self- 
administration, many studies have been focused on the dopamine transporter. A highly detailed 
structure- activity study of the binding of cocaine analogues to the dopamine transporter is being 
undertaken. At this point, approximately 60 cocaine analogues have been screened in binding assays. 
Several compounds that are an order of magnitude more potent that any other known cocaine analogue 
have been identified. In addition, several irreversible binding probes have been synthesized and 
characterized. The carbohydrate moiety of the dopamine transporter has been characterized extensively 
using enzymes that remove specific sugars from glycoproteins. It has been shown that the carbohydrate 
moiety is rich in sialic acids. Efforts are also underway to purify the dopamine transporter. 

B. Drug Receptors, Neurotransmitters and Addiction. 

This is a multifaceted project aimed at examining a variety of drugs and neurotransmitters that are 
associated with addiction. In one of our studies, we examined the binding of PCP to striatal cocaine 
receptors. We found that PCP has a low affinity for cocaine receptors and therefore would interact with 
cocaine receptors only at high concentrations. Thus, the bulk of the psychotropic effects of PCP are 
unlikely to be due to interaction with cocaine receptors. We also characterized the binding of opiate 
receptors with a new ligand, carfentanii, and identified a new ligand, spectramide, for D2 dopamine 
receptors. Several studies are underway examining the effects of drug administration on dopamine 
receptors in the brain. 

C. Measuring Drug Receptors In Vivo 

Several new in vivo labeling ligands for the dopamine transporter have been identified. Perhaps the most 
promising is radiolabeled WDM 35,428. This ligand binds to cocaine receptors in vivo more efficiently 
than any other known compound. This finding allows us to study the occupancy of cocaine receptors in 
vivo by a variety of compounds. For example, in preliminary studies we recently showed that mazindol 
enters the brain and occupies cocaine receptors much more slowly than cocaine itself. These in vivo 
labeling studies wdl justify a study of cocaine receptors in living humans by PET and SPECT scanning in 
the near future. 



Publications 

Kuhar. M.J.. P.M. Sanchez-Roa. D.F. Wong, R.F. Dannais. D.E. Grigoriadis. R. Lew and M. Milberger. 
Dopamine Transporter: Biochemistry, Pharmacology and Imaging. Eur. Neurol. 30(1). 15-20. 1990. 

Ritz, M.C., E.J. Cone and M.J. Kuhar. Cocaine Inhibition of Ligand Binding at Dopamine. 
Norepinephrine and Serotonin Transporters: A Structure- Activity Study. Life Sciences 46_, 635-645, 
1990. 

Kuhar, M.J. and J.R. Unnerstall. Receptor Autoradiography. In: Methods in Nenrotransmitter Receptor 
Analysis . I. Yamamura et. al. (Eds.) Raven Press, Ltd., New York. pp. 177-218, 1990. 

Kuhar. M.J. Introduction to Neurotransmitters and Neuroreceptors. In: Quantitative Imaging: 
Neuroreceptors. Neu rotransmi tters, and Enzvmes . J. James Frost and Henry W. Wagner, Jr. (Eds.) 
Raven Press. Ltd. New York. pp. 1-7. 1990. 

Kuhar, M.J., J.W. Boja and E.J. Cone Phencyclidine Binding to Striatal Cocaine Receptors. 
Neuropharmacology 29: 295-297, 1990. 

Boja, J.W., F.I. Carroll, M. Abdur Rahman, A. Philip, A.H. Lewin and M.J. Kuhar. New, Potent Cocaine 
Analogs: Ligand Binding and Transport Studies in Rat Striatum. Eur. J. Pharmacol. 154:329-332, 1990. 

Kuhar, M.J. Cocaine Receptors and Dopamine Transporters. Neurosci. Facts 1(3):3, 1990. 

Zarbin, M.A., J.K. Wamsley and M.J Kuhar. Anterograde Transport of Opioid Receptors in Rat Vagus 
Nerves and Dorsal Roots of Spinal Nerves: Pharmacology and Sensitivity to Sodium and Guanine 
Nucleotides. Ex p. Brain Research 81: 267-278. 1990. 

Naseree, T.M.. P. Abraham, J.A. Kepler, F.L Carroll, A.H. Lewin and M.J. Kuhar. Synthesis of 

[ 3 H|WIN 35.065-2; A New Radioligand for Cocaine Receptors. Journal of Labelled Compounds and 
Radiopharmaceuticals. 28(9): 101 1-1016. 1990. 

Kuhar. M.J. A GAB A Transporter cDNA Has Been Cloned. Trends Neurosci. 13:473-474. 1990. 

Ritz, M.C., J.W. Boja, D. Grigoriadis, R. Zaczek. F.I. Carroll, A.H. Lewis and M.J. Kuhar. [ 3 H]WIN 
35,065-2: A ligand for cocaine receptors in striatum. J. Neurochem. 55. 1556-1562, 1990. 

Boja, J.W. and Schechter, M.D. Increased drug sensitivity in the drug discrimination procedure afforded 
by drug vs. drug training. Psvchopharmacologv 102 :221-226. 1990. 

Kuhar, M.J. Cocaine receptors and dopamine transporters. Neurosci. Eacjs 1(3):3, 1990. 

Kuhar. M.J. Introduction to neurotransmitters and neuroreceptors. In: Quantitative imaging: 
Neuroreceptors, neurotransmitters and enzvmes. J. James Frost and Henry J. Wagner, Jr. (Eds.) Raven 
Press. Ltd., New York. pp. 1-7, 1990. 

Articles in Press 

Lew, R., Grigoriadis, D., Wilson, A., Boja, J.W., Simantov, R. and Kuhar. M.J. Dopamine transporter: 
Deglycosylation with exo- and endo-glycosidases. Accepted, Brajn Research. 1990. 



Boja, J.W., Rahman, M.A.. Philip, A.. Lewin, A.H., Carroll. F.L and Kuhar. M.J. Isothiocyanate 
derivatives of cocaine: Irreversible of ligand binding at the dopamine transporter. Accepted. Mol. 
Pharmacol. . 1990. 

Carroll, F.L. Lewin, A.H., Philip. A., Parham, K.. Boja, J.W. and Kuhar. M.J. Synthesis and ligand 
binding of cocaine isomers at the cocaine receptor. Accepted. J. Med Chem. . 1990. 

Cline. E.J.. Scheffel. U., Boja. J.W.. Carroll. F.L. Katz. J.L. and Kuhar, M.J. Behavioral effects of novel 
cocaine analogs: a comparison with in vivo receptor binding potency. Submitted. J. Pharmacol. Exp. 
Ih£L, 1990. 

Kuhar. M.J. Neuropeptides in the CNS, Part II. In: Handbook of Chemical Neuroanatomy . A. 
Bjorklund and T. Hokfelt (Eds). Elsevier. New York. Vol. 9, 1990. 

Vaughan. R.A.. Simantov, R., Lew. R. and Kuhar. M.J. A rapid binding assay for solubilized dopamine 
transporters using 13H]-WIN 35.428. Submitted. J. Neurosci. Methods . 1990. 

Pogun, S., Scheffel. U.. and Kuhar. M.J. Cocaine binds to dopamine uptake sites in vivo more rapidly 
than mazmdol or GBR 12909. Submitted. Eur. J. Pharmacol. . 1990. 

Scheffel. U.. Pogun. S.. Stathis. M., Boja, J.W. and Kuhar. M.J. In vivo labeling of cocaine binding sites 
on dopamine transporters with [3H]WIN 35,428. Submitted. J. Pharmacol. Exp. Ther. . 1990. 

Battaglia. G., Sharkey, J., Kuhar, M.J., and De Souza. E.B. Neuroanatomic specificity and time course 
of alterations in rat brain serotonergic pathways induced by MDMA (3,4-methylenedioxy- 
methamphetaminet: Assessment using quantitative autoradiography. Submitted. Synapse . 1990. 

Simantov. R., Vaughan. R., Lew, R.. Wilson. A. and Kuhar. M.J. Dopamine transporter - cocaine 
receptor: Characterization and purification. Submitted. Advances in the Biosciences . 1990. 

Lew, R., Vaughan. R.. Simantov. R., Wilson, A. and Kuhar. M.J. Dopamine transporters in the nucleus 
accumbens and the striatum have different apparent molecular weights. Submitted. Svnapse . 1990. 

Abstracts Published 

Lew, R., J.W. Boja, A. Wilson and M.J. Kuhar. Identification of the Dopamine Transporter in Various 
Regions of the Rat Brain Using the Photoaffinity Radioligand [ 125 I]DEEP. FASEB Journal 4(3), 342. 
1990. 

Carroll. F.L, J.W. Boja, A. Philip, A.H. Lewin. and M.J. Kuhar. Structure-Activity Relationships of Ring 
and Side-group Substitutions of the Cocaine Molecule. FASEB Journal 4(3), 343, 1990. 

Boja, J.W., A. Lewin. F.L Carroll and M.J. Kuhar. Irreversible Binding to the Cocaine Receptor by 
Meta- and Para-isothiocyanato- benzoylecgonine Methyl Ester. FASFB Jnnmal 4(3), 344, 1990. 

Scheffel, U., J.W. Boja, M. Stathis and M.J. Kuhar. In Vivo Labeling of Cocaine Receptors With 3 H-(- 
)Cocaine. 3 H-WIN 35.065-2 and 3 H-WIN 35.428. FASEB Journal 4(3), 2781, 1990. 

Vaughan. R.A., R. Simantov, R. Lew, E. Webster and M.J. Kuhar. A Rapid Binding Assay for Soluble 
Dopamine Transporters. Soc. Neurosci. 16 (1). 745, 1990. 

Scheffel, U., R.F. Dannals, A.A. Wilson, H.T. Ravert, J.W. Boja, M. Stathis and M.J. Kuhar. 3H/11C- 
WIN-35,428 Labels the Cocaine Receptor In Vivo . Soc. Neurosci. 16 ( 1), 746, 1990. 



Lew, R., J.W. Boja, R. Simantov, F.I. Carroll, A. Lewin and M.J. Kuhar. New Photaffinity Probes for 
the Cocaine Receptor. Soc. Neurosci. 1£ (1), 746, 1990. 

Boja, J.W., T. Kopajtic. F.I. Carroll, A. Lewin and M.J. Kuhar. High Affinity Inhibition of the Cocaine 
Binding Site by Novel Cocaine Analogs. Soc. Neurosci. 16 ( 1), 746, 1990. 

Boja, J.W.. E.J. Cline. D.M. Pearsall, F.I. Carroll, A. Lewin and M.J. Kuhar. Neurochemical and 
Behavioral Effects of Novel High Potency Cocaine Analogs. Presented at ACNP Meeting, San Juan, 
Puerto Rico, 1990. 

Kuhar, M.J.. F.I. Carroll. U. Scheffel, R. Dannals, D. Wong, E. Shaya and J.W. Boja. New Tracers and 
the Distribution of (-) Cocaine Binding Sites in the Brain. Presented at ACNP Meeting, San Juan, Puerto 
Rico. 1990. 

Boja, J.W., F.I. Carroll and M.J. Kuhar. The Dopamine Transporter Binding by the Irreversible ligands 
meta- and para-isothiocyanato-benzylecgonine methyl ester. Presented at the 11th IUPHAR Congress 
Amsterdam, The Netherlands. 1990. 

Simantov. R.. R. Lew, J.W. Boja, D.E. Grigonadis, R. Zaczek. R. Vaughan and M.J. Kuhar. Dopamine 
Transporter as a Cocaine Receptor. IUPHAR Satellite International Symposium on Presynaptic 
Receptors & Neuronal Transports. Rouen, France, 1990. 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00112-04 MPL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: Drug Receptors, Neurotransmitters and Addiction 

Principal Investigators: Cooperating Units 

P.I. M.J. Kuhar Chief, Neuroscience Branch 

Others: Cone, E. Chief, Chemistry Laboratory, Clinical Pharmacology Branch 

Boja, J.W. Staff Fellow, ARC 

Cooperating Unit: None 

Lab/Branch: Laboratory of Molecular Pharmacology, 

Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 1 Professional: 3/4 Other: 1/4 

Check Appropriate Boxes: 

Human Subjects Human Tissues _2£ Neither 

Minors 

Interviews 

Summary of Work 

Tliis project focuses on various drugs of abuse and the receptors with which they interact. We completed 
a study examining the binding of PCP and related compounds at the cocaine receptor. The relative 
potencies of these compounds were such that some of the behavioral effects of PCP might be related to 
its action at the cocaine receptor, in other words, the affinity of PCP at the cocaine binding site was 
interesting but considerably less than its affinity at its own receptor. (Kj of 1.59 mM vs. a Kj of about 

0.12 mM). Thus, at high blood levels, PCP could interact with the cocaine receptor but it would most 
likely occupy the PCP site at the NMDA receptor at much lower concentrations. 

Another study examined the movement of opiate receptors within neurons. Receptors are synthesized by 
the usual protein synthetic machinery of cells. They are then transported to sites and inserted into the 
membrane. We examined the transport of opiate receptors in rat vagus and in dorsal roots of spinal 
nerves. We found that the opiate receptors were transported in these neurofibers by fast transport and this 
agrees with many studies of other receptors. The binding sites in the axons had properties similar to 
those for receptors observed in brain tissue; the binding sites had appropriate pharmacology, were GTP 
sensitive as well as showed effects of sodium. 



PUBLICATIONS 

Kuhar, M.J.. J.W. Boja and E.J. Cone Phencyclidine Binding to Striatal Cocaine Receptors. 
Neuropharmacology 29: 295-297, 1990. 

Zarbin, M.A.. J.K. Wamsiey and M.J Kuhar. Anterograde Transport of Opioid Receptors in Rat Vagus 
Nerves and Dorsal Roots of Spinal Nerves: Pharmacology and Sensitivity to Sodium and Guanine 
Nucleotides. Exp. Brain Research Si: 267-278, 1990. 

Kuhar, M.J. A GABA Transporter cDNA Has Been Cloned Trends Neurosci. 12:473-474, 1990. 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00107-05 MPL 



Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Drug Receptors In Vivo 

Principal Investigators: Cooperating Units 

P.I. M.J. Kuhar Chief, Neuroscience Branch 

Others: Wong, D. Division of Nuclear Medicine, JHU 

Wagner. H.N. Division of Nuclear Medicine, JHU 

Scheffel, U. Division of Nuclear Medicine, JHU 

Dannals, R. Division of Nuclear Medicine. JHU 

Cline. E. PRAT Fellow, Neuroscience Branch 

Pogun S. Division of Nuclear Medicine. JHU 

Shaya, E. Division of Nuclear Medicine, JHU 

Cooperating Unit: Division of Nuclear Medicine. 

Johns Hopkins University School of Medicine 

Lab/Branch: Laboratory of Molecular Pharmacology, 

Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3 Professional: 2 1/4 Other: 3/4 

Check Appropriate Boxes: 

Human Subjects Human Tissues _X_ Neither 

Minors 

Interviews 

Summary of Work 

Considerable progress has been made in the area of imaging cocaine receptors in living animals. Our 
first efforts involved utilizing a known antidepressant drug, nomifensine. A detailed study of the in vivo 
binding characteristics of this compound revealed that it localized to dopamine transporters quite 
effectively but with a low specific to non-specific binding ratio. Because of this, we set out to develop 
more efficient binding ligands. 

Studies from our laboratory previously showed that cocaine analogs, WTN-35,065-2 and WIN-35,428 
were better ligands for the dopamine transporter in that higher specific to non-specific binding ratios 
were obtained in vivo . More recendy, we utilized WIN-35,428 in a more extensive study which 
conclusively showed that this compound localized to cocaine receptors/dopamine transporters in rat 
striatum. We are in the process of utilizing this compound in its positron emitting form to carry out PET 
scanning studies of dopamine transporters in vivo . 

One important use of in vivo labelling techniques is to examine the rate of occupancy of receptors in vivo 
by various drugs. Because of the avadability of our in vivo labeling model using WIN-35,428, we were 
able to examine the relative rate of occupancy of cocaine receptor by mazindol. GBR- 12,909 and 



cocaine. We clearly showed that cocaine enters the brain and occupies cocaine receptors much more 
rapidly than mazindol or GBR-1 2,909. It is known that abuse liability is greater for drugs that enter the 
brain and occupy receptors rapidly. Thus, this model will be useful in quantitatively assessing how 
rapidly drugs enter the brain and occupy receptors; this will be helpful in contributing quantitative 
knowledge of this factor for abuse liability assessment. 

PUBLICATIONS 

Kuhar. M.J., P.M. Sancbez-Roa, D.F. Wong, R.F. Dannals. D.E. Grigoriadis. R. Lew and M. Milberger. 
Dopamine Transporter: Biochemistry, Pharmacology and Imaging. Eur. Neurol. 30(1) . 15-20, 1990. 

Pogun, S., Scheffel, U., and Kuhar, M.J. Cocaine binds to dopamine uptake sites in vivo more rapidly 
than mazindol or GBR 12909. Eur. J. Pharmacol. , in press. 

Scheffel, U., Pogun. S., Stathis, M., Boja, J.W. and Kuhar, M.J. In vivo labeling of cocaine binding sites 
on dopamine transporters with PH]WIN 35,428. J. Pharmacol. Exp. Ther. . in press. 

ABSTRACTS 

Scheffel, U.. J.W. Boja, M. Stathis and M.J. Kuhar. In Vivo Labeling of Cocaine Receptors With %-(- 
(Cocaine. 3 H-WIN 35.065-2 and 3 H-WIN 35,428. FASEB Journal 4(3). 2781, 1990. 

Scheffel. U., R.F. Dannals, A.A. Wilson, H.T. Ravert, J.W. Boja,M. Stathis and M.J. Kuhar. 3H/11C- 
WDM- 3 5 .428 Labels the Cocaine Receptor In Vivo . Soc. Neurosci, 16(1), 746, 1 990. 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00108-04 MPL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: The Cocaine Receptor 
Principal Investigators: Cooperating Units 



P.I. 


M.J. Kuhar 


Chief, Neuroscience Branch 


Others: 


Boja, J.W. 


Staff Fellow, ARC 




Lew, R. 


Visiting Fellow, ARC 




Simantov, R. 


Visiting Fellow. ARC 




Carroll, I. 


Research Triangle Institute 




Vaughan, R. 


Visiting Fellow. ARC 



Cooperating Unit: RTI 

Lab/Branch: Laboratory of Molecular Pharmacology, 

Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse. Baltimore. MD 21224 

Total Man Years: 6 Professional: 5 Other: 1 

Check Appropriate Boxes: 

Human Subjects Human Tissues _X_ Neither 

Minors 

Interviews 

Summary of Work 

In order to carry out a detailed study of the cocaine receptor, we have synthesized and tested a large 
number of cocaine analogs in collaboration with Dr. Ivy Carroll at RTI. For example, we found that 
analogs of cocaine that are also analogs of WIN 35,428 are potent in ligand binding studies as well as in 
dopamine transport studies in rat striatal tissue. These analogs are the most potent cocaine analogs 
known, some of them being 80 to 100 times more potent than cocaine itself. We have also synthesized 
3H-WIN 35,065-2 and carried out binding studies because it is a potent cocaine compound that has been 
studied by a variety of laboratories. 

These compounds also contribute substantially to structure- activity relationship information. We have 
shown that halogen substituents on the phenyl ring at the C3 carbon can increase potency. These studies 
have implications for understanding the nature of the cocaine receptor in the binding region of the 
molecule. 

We have also utilized irreversible binding ligands at the cocaine receptor/dopamine transporter to study 
the carbohydrate moiety of the protein molecule. Our results indicate that the carbyhydrate is rich in 
sialic acid residues and is probably N-linked to the protein. 



10 



PUBLICATIONS 

Ritz. M.C.. E.J. Cone and M.J. Kuhar. Cocaine Inhibition of Ligand Binding at Dopamine. 
Norepinephrine and Serotonin Transporters: A Structure-Activity Study. Life Sciences 46_, 635-645, 
1990. 

Boja, J.W., F.I. Carroll, M. Abdur Rahman. A. Philip, A.H. Lewin and MJ. Kuhar. New, Potent Cocaine 
Analogs: Ligand Binding and Transport Studies in Rat Striatum. Eur. J. Pharmacol. 124:329-332, 1990. 

Kuhar, M.J. Cocaine Receptors and Dopamine Transporters. Neurosci. Facts 1(3):3, 1990. 

Naseree, T.M., P. Abraham, J.A. Kepler, F.L Carroll, A.H. Lewin and M.J. Kuhar. Synthesis of 

[%|WIN 35.065-2: A New Radioligand for Cocaine Receptors. Journal of Labelled Compounds and 
Radiopharmaceuticals . Vol. XXVUI . No. 9: 1011-1016, 1990. 

Ritz, M.C., J.W. Boja, D. Grigoriadis, R. Zaczek, F.I. Carroll, A.H. Lewis and M.J. Kuhar. [ 3 H]WIN 
35,065-2: A ligand for cocaine receptors in striatum. J. Neurochem. 5_5_, 1556-1562, 1990. 

Lew, R., D. Grigoriadis, A. Wilson, J.W. Boja, R. Simantov and M.J. Kuhar. Dopamine transporter: 
Deglycosylation with exo- and endoglycosidases. Brain Research , in press. 

Boja, J.W., MA. Rahman. A. Philip, A.H. Lewin, F.I. Carroll and M.J. Kuhar. Isothiocyanate 
derivatives of cocaine: Irreversible inhibition of ligand binding at the dopamine transporter. Mol. 
Pharmacol. , in press. 

Carroll. F.L, A.H. Lewin. A. Philip, K. Parham, J.W. Boja and M.J. Kuhar. Synthesis and ligand binding 
of cocaine isomers at the cocaine receptor. J. Med. Chem. . in press. 

Boja, J.W.. A. Patel, F.I. Carroll. MA. Rahman, A. Philip, A.H. Lewin and M.J. Kuhar. [ 125 I]RTI-55: 
A potent ligand for dopamine transporters. Eur. J. Pharmacol. , in press. 



ABSTRACTS 

Lew, R., J.W. Boja, A. Wilson and M.J. Kuhar. Identification of the Dopamine Transporter in Various 
Regions of the Rat Brain Using the Photoaffinity Radioligand [ 125 I]DEEP. FASEB Journal 4(3), 342, 
1990. 

Carroll, F.L, J.W. Boja, A. Philip, A.H. Lewin, and M.J. Kuhar. Structure-Activity Relationships of Ring 
and Side-group Substitutions of the Cocaine Molecule. FASEB Journal 4(3), 343, 1990. 

Boja, J.W., A. Lewin, F.I. Carroll and M.J. Kuhar. Irreversible Binding to the Cocaine Receptor by 
Meta- and Para-isothiocyanato- benzoylecgonine Methyl Ester. FASEB Journal 4(3). 344, 1990. 

Vaughan, R.A., R. Simantov, R. Lew, E. Webster and M.J. Kuhar. A Rapid Binding Assay for Soluble 
Dopamine Transporters. Soc. Neurosci. 16 (1), 745, 1990. 

Lew, R., J.W. Boja, R. Simantov, F.I. Carroll, A. Lewin and M.J. Kuhar. New Photaffinity Probes for 
the Cocaine Receptor. Soc. Neurosci. 16 (1), 746, 1990. 

Boja, J.W., T. Kopajtic, F.I. Carroll, A. Lewin and M.J. Kuhar. High Affinity Inhibition of the Cocaine 
Binding Site by Novel Cocaine Analogs. Soc. Neurosci. 16 (1), 746, 1990. 



11 



2. Neuropharmacology Laboratory -- Edythe D. London. Ph.D., Chief 

Overview 

The Neuropharmacology Laboratory investigates biological systems that mediate effects of abused drugs. 
Studies focus on molecular mechanisms, involving neurotransmitter systems. Techniques used include 
receptor binding, purification and identification of endogenous neuroactive substances, 
electrophysiology, and brain imaging. The information obtained may help design new treatments, 
including medications for substance abuse. 

Summary of Ongoing Research: 

A. Brain Imaging 

These studies aim to elucidate brain mechanisms that underlie the etiology and sequelae of drug abuse. 
Euphorigenic treatments with morphine or cocaine reduced the regional cerebral metabolic rate for 
glucose (rCMRglc) in humans, particularly in cortical areas. These are the first and only large-scale 
studies with street-equivalent doses of euphoriant drugs. Cerebral ventnculomegaiy was associated with 
insensitivity to the euphorigenic and cardiac stimulant properties of cocaine. The studies are being 
continued to relate mood to rCMRglc. Volunteers without drug abuse histories are being studied to 
determine if substance abusers have abnormal brain metabolism or electrical activity. A new study is 
examining effects of nicotine, with respect to tolerance and metabolic correlates of euphoria. Other 
protocols have been initiated to examine opioid and cocaine withdrawal, and potential therapeutic 
interventions. Methods to improve quantitation of PET measurements are in development. 

Studies in rodents have focused on nicotine, psychomotor stimulants, and GP120 (HIV envelope 
glycoprotein). Chronic nicotine treatment produced tolerance to nicotine's effects on rCMRglc. 
Psychomotor stimulants, including cocaine and methylenedioxymethamphetamine (MDMA) stimulated 
rCMRglc in the extrapyramidal motor system and reduced rCMRglc in the lateral habenula. Findings 
with MDMA were consistent with stimulant and hallucinatory actions. Genetic differences in 
responsivity to cocaine were observed. As drug abusers are at risk for HIV infection, we examined the 
effect of GP120 on rCMRglc. GP120 reduced brain metabolism, reminiscent of decrements observed in 
AIDS patients with dementia. Studies in mice were performed to develop new ligands for imaging 
receptors with PET. The results demonstrated the feasibility of using radiolabeled nicotine and 
halopendol as in vivo ligands. 

B. Physiological Effects of Opioids 

This project centers on neural systems that contribute to opioid effects. Interactions with Ca2+ 
antagonists were tested. In human volunteers, verapamil antagonized morphine-induced respiratory 
depression and tended to block euphoria. Interactions between nifedipine and U50488H on activities of 
L- and N-type Ca2+ channels in cultured dorsal root ganglion cells suggested additive effects at L and N 
channels. In the isolated spinal cord, nifedipine inhibited long latency C-fiber reflexes, but did not affect 
monosynaptic reflexes. Kappa and 8 agonists had similar effects. Such studies may lead to safer and 
more effective opioid treatments. 

Studies of the opioid abstinence syndrome involve intact rats, isolated neonatal rat spinal cords, and co- 
cultures of dorsal root ganglion and spinal dorsal hom neurons. Cerebral metabolic studies in rats 
showed that clonidine attenuates hypermetabolism during morphine withdrawal. We identified limbic 
and hypothalamic areas, not previously implicated in these phenomena. 

Isomers of ketocyclazocine were tested for their ability to produce k and o effects in the dog. The 
inactivity of d- vs. 1-ketocyclazocine did not reflect pharmacokinetics, and the actions of 1- 



12 



ketocyclazocine typically were K-like, lacking pbencyclidine- or o-like activity. We demonstrated that 
BW942C is a peptidic partial agonist at k opioid receptors. Naltrexone-induced salivation in rats was 
established as a model for involvement of various opioid receptors in behavioral conditioning. 1- 
Ketocyclazocine mainly acted as a k drug whereas no pharmacological actions of d-ketocyclazocine were 
seen. We are investigating the mechanism by which DADLE. a 6 opioid receptor ligand, prolongs organ 
survival time in an autoperfusion multi-organ preparation. 

C. Ligand-Gated Ion Channels 

This project is directed at elucidating modulatory mechanisms that govern the function of the superfamily 
of ligand-gated ion channels. The family includes the N-methyl-D- aspartate (NMDA), the nicotinic, and 
the gamma-aminobutyric acid (GABA) receptors. 

The NMDA receptor has been a focus of study because of the interaction of pbencyclidine ( PCP) with 
sites inside the open state of the cationic channel formed by this receptor. We have studied new 
dimensions of NMDA receptor function, by identifying and characterizing polyamine binding sites on the 
NMDA receptor complex, and by discovering a mechanism of NMDA receptor regulation by redox 
phenomena. Some reductants (ascorbic acid, hydroquinone > inhibit ligand binding and receptor function, 
and protect cultured cortical cells from neurotoxicity due to exitatory amino acids. Others (dithiothreitol, 
mercaptoethanol ). which break cystine disulfide bonds, potentiate NMDA action. Polyamine receptors 
offer promise of providing a template for the design of new therapeutic agents directed at the function of 
the NMDA receptor. 

To understand the molecular basis of nicotine addiction, we study endogenous mechanisms regulating the 
function of nicotinic cholinergic receptors (nAChRs). We characterized interactions of noncompetitive 
inhibitors of nAChRs in rat brain, and used [3H]mecamylamine as a new radioligand probe for the 
cationic channel of nAChRs. Nucleotides, in particularly ATP, were modulators of binding both at 
acetylcholine recognition sites and at sites thought to be located within the channel, suggesting a 
mechanism for modulation of nAChR function. 

We investigated the actions of substances which might influence GABAa receptors, which are primary 
targets for the actions of benzodiazepines. By using [3H]pregnenolone sulfate and 

[3H]dehydroepiandrosterone sulfate, modulatory sites on the GABAa receptor complex were identified 
and characterized. Electrophysiological studies demonstrated that DHEAS is a noncompetitive inhibitor 
of the GABAa receptor. 

D. Sigma (o) Receptor 

The purpose of this project is to discover the biological roles of o receptors. Structure-activity studies 
identified the most selective a ligand (PRE-084) reported to date. Neuronal a receptors were found 
throughout the animal kingdom. In cerebral metabolic studies, o drugs elicited responses in o receptor 
dense areas. Sigma receptors are selectively lost in certain areas of schizophrenic brains. A low-affinity 
receptor, closely related to tonic K+ channels was identified. The report was the first of a receptor 
affecting such a channel. The possible relationship between high- and low-affinity o receptors is being 
investigated. Sigma receptors displayed a unique subcellular distribution. SolubUized receptors from rat 
brain and liver retain all pharmacological characteristics of membrane-bound receptors, including high 
affinity for progesterone. Purification of o receptors is underway. 



13 



Publications 

Vu. T.H.. Weissman, A.D. and London, E.D. Pharmacological characteristics and distributions of a and 
pbencyclidine binding sites in the animal kingdom. J. Neurochem. 54:598-604, 1990. 

London, E.D.. Broussolle, E.P.M.. Links, J.M., Wong, D.F., Cascella, N.G., Dannals, R.F., Sano, M.. 
Herning, R., Snyder, F.R., Rippetoe, L.R., Toung, T.J.R., Jaffe, J.H. and Wagner, H.N., Jr.: Morphine- 
induced metabolic changes in human brain - studies with positron emission tomography and [fluorine 
181fluorodeoxyglucose. Arch. Gen. Psvchiat. 47:73-81. 1990. 

Vaupel, D.B., Cone, E.J., Johnson, R.E., and Su, T.-P. Kappa opioid partial agonist activity of the 
enkephalin-like pentapeptide BW942C based on urination and in vitro studies in humans and animals. L 
Pharmacol. Exp. Ther . 252:225-234, 1990. 

Su, T.-P., and Wu, X.-Z. Guinea-pig vas deferens contains a but not phencyclidine receptors. Neurosci. 
Lett. 108:341-345, 1990. 

Schindler. C.W.. Wu, X.-Z., Su, T.-P., Goldberg, S.R., and Katz, J.L. Enhanced sensitivity to the 
behavioral effects of naltrexone in rats: A conditioning phenomenon associated with opioid receptor 
changes. J. Pharmacol. Exp. Ther. 252:8-14. 1990. 

Su, T.-P., Chien, S.F. and Oeltgen, P.R. Hibernation induction trigger (HIT) extends preservation time in 
an autoperfusion multiorgan preparation. Clin. Pharmacol. Ther. 47:140. 1990. 

Kimes, A.S., Bell, J. A., and London, E.D. Clonidine antagonizes increased glucose metabolism during 
naloxone-precipitated morphine withdrawal. Neuroscience 34 (3):633-644. 1990. 

London, ED., Wilkerson, G.W., Ori, C. and Kimes, A.S.: Central action of psychomotor stimulants on 
glucose utilization in extrapyramidal motor areas of the rat brain. Brai n Res , 5_L2:155-158, 1990. 

Henningfield, J.E., London, E.D. and Benowitz, N.L.: Arterio-venous differences in plasma 
concentrations of nicotine after cigarette smoking. JAMA 263 :2049-2050, 1990. 

McCann. D.J.. and Su. T.-P.: Halopendol-sensitive <+)[ 3 H]SKF-10,047 binding sites ("a sites") exhibit a 
unique distribution in rat brain subcellular fractions. Eur. J. Pharmacol . 188 :21 1-218, 1990. 

London, E.D., Cascella, N.G., Wong, D.F.. Phillips, R.L., Dannals. R.F., Links, J.M., Herning, R., 
Grayson, R., Jaffe, and Wagner, Jr., H.N.: Cocaine-induced reduction of glucose utilization in human 
brain: A study using positron emission tomography and [fluorine 18]fluorodeoxyglucose. Arch. Gen. 
Psvchiat . 47:567-574, 1990. 

Mantione, C.R., Demirgoren, S. and London, E.D.: Specific binding of [3H]spermidine to membranes of 
rat brain. Eur. J. Pharmacol. 180 :393-394. 1990. 

London. E.D.: Effects of nicotine on cerebral metabolism. In: The Biology of Nicotine Dependence 
(Ciba Foundn. Symp. No. 152), (G. Bock and J. Marsh, eds.) Wiley, Chichester, pp. 131-146, 1990. 

McCann, D.J., and Su, T-.P.: Haloperidol competitively inhibits the binding of (+)[3H]SKF- 10,047 to o 
sites. Eur. J. Pharmacol . !8J}:361-364, 1990. 

London, E.D., Fanelli, R.J., Kimes, A.S. and Moses, R.L.: Effects of chronic nicotine on cerebral 
glucose utilization in the rat. Brain Res. 520 :208-214, 1990. 



14 



Bell, J. A.: Naloxone-induced facilitation of C-fiber reflexes is reduced by chronic morphine. Eur. J. 
Pharmacol. 168 :101-105. 1990. 

Majewska. M.D., Demirgoren, S., Spivak, C.E. and London. E.D.: The neurosteroid 
dehydroepiandrosterone sulfate is an allosteric antagonist of the GABA^ receptor. Brain Res 5_26_:143- 

146. 1990. 

Majewska. M.D.: Steroid regulation of GABA receptor: ligand binding, chloride transport, behavior. In: 
Steroids and p^in Activity (Ciba Foundn. Symp. No. 153), (G. Bock and J. Marsh, eds.) Wiley, 
Chichester, 1990. PP. 83-106. 

Weissman, ED., Broussolle. E.P. and London, E.D.: In vivo binding of [3H]d-N-allylnormetazocine and 
[3H")haloperidol t0 sigma receptors in the mouse brain. J. Chem. Neuroanatomy 1:347-354, 1990. 

Gund. T.M., Shukla. K. and Su, T-.P.: A unifying hypothesis for binding of psychotomimetic ligands at 
the sigma receptor. Chem. Pes. Autom. News 5:1-20. 1990. 

Majewska. M.D., Demirgoren, S. and London. E.D.: Binding of pregnenolone sulfate to rat brain 
membranes suggests multiple sites of steroid action at the GABA^ receptor. Eur. J. Pharmacol. . Mol. 

Pharmacol. Section 189 :307-315. 1990. 

Majewska. M.D.: Neurosteroids, metabolism and function. Neuroscience Facts 1:2, 1990. 

Majewska, M.D., Bell, J.A. and London, E.D.: Regulation of the NMDA receptor by redox phenomena: 
Inhibitory role of ascorbate. Brain Res. 537:328-332. 1990 

Phillips, R.L., London. E.D., Links, J.M., Cascella, N.G.: Program for PET image alignment: Effects on 
calculated differences in cerebral metabolic rates for glucose. J. Nucl. Med. 3.1:2052-2057, 1990. 

Majewska. M.D. and Bell, J.A.: Ascorbic acid protects neurons from injury induced by glutamate and 
NMDA. Neuroreport 1:194-196, 1990. 

London. E.D.: Glucose metabolism: An index of nicotine action in the brain. In: Proceedings of 
International Symposium on Nicotine. June 28-30, 1990, Hamburg. Advances in Pharmacological 
Sciences . (A. Adlkofer, Ed.), Birkhauser Verlag, Basel, 1990. 

Morgan, M.J. and Franklin, K.B.J. : 6-Hydroxydopamine lesions of the ventral tegmentum abolish D- 
amphetamine and morphine analgesia in the formalin test but not in the tail flick test. Brain Res . 
5J9_:144-149, 1990. 

ARTTC1.FS TN PRESS 

London, E.D.: Studies of a receptors and metabolic responses to o ligands in the brain. In: Si gma. PCP 
and NMDA Receptor Systems . EB De Souza, ED London, and D Clouet, eds. NBDA Research 
Monographs. 

Su, T-.P.: Pharmacological characterizations of o receptors. In: Si gma. PCP and NMD A Receptor 
Systems . NTDA Research Monographs. EB De Souza, ED London, and D Clouet, eds. DHHS. NJJDA. 

Su, T-.P., Shuklar, K. and Gund, T.: Steroid binding at receptors: CNS and immunological 
implications. In: Steroids and Neuronal Activity (Ciba Foundn. Symp. No. 153), (D.J. Chadwick, K. 
Widdows, eds.) Wiley, Chichester, 1990. 



15 



London, E.D., Morgan, M.J.. Phillips, R.L., Stapleton, J.M., Casceila, N.G. and Wong, D.F.: Mapping 
the metabolic correlates of drug-induced euphoria. NIDA Research Monograph : Proceedings of the 
Committee on Problems of Drug Dependence 1990. 

Weissman, A.D., Cassanova, M.F., Kleinman, J.K., London, E.D. and De Souza. E.B.: Selective 
reduction of cerebral cortical sigma. but not PCP binding sites in schizophrenia. Biol. Psvch . 

Cohen, S.R., Kimes, A.S. and London, E.D.: Morphine decreases cerebral glucose utilization in limbic 
and forebrain regions while pain has no effect. Neuropharmacology . 

London, E.D., Dawson, V.L. and Mantione, C.R.: Specific binding sites for polyamines in mammalian 
brain. In: NMDA Related Agents: Biochemistry. Pharmacology and Behavior . Proceedings from a 
satellite symposium of the 17th Congress of Collegium Internationale Neuro-Psychopharmacologicum. 
Nagoya, Japan, Sept. 16-17, 1990, NPP Books. Inc., Ann Arbor, Ml. T. Nabeshima. T. Kameyama. and 
E.F. Domino, eds. 

Spivak. C.E., Waters. J. A. and Aronstam. R.S.: t+)-Octahydro-2- methyl- trans -5( IHVisoquinolone 
methiodide: A probe to test the stenc limits of agonists active at the nicotinic acetylcholine receptor. J. 
Mol. Graphics . 

Su, T-.P., Wu, X-.Z..Spivak, C.E., London, E.D. and Bell, J.A.: Binding studies on intact NCB-20 cells 
suggest o receptor multiplicity: c\ and 02. In: NMDA Related Agents: Biochemistry, Pharmacology 
and Behavior . Proceedings from a satellite symposium of the 17th Congress of Collegium Internationale 
Neuro-Psychopharmacologicum, Nagoya, Japan, Sept. 16-17, 1990, NPP Books, Inc., Ann Arbor, MI. T. 
Nabeshima, T. Kameyama, and E.F. Domino, eds. 

Kimes, A.S., London, E.D., Szabo, G., Raymon. L. and Tabakoff, B.: Reduction of cerebral glucose 
utilization by the HTV envelope glycoprotein gp- 1 20. Exp. Neurol. 

Vaupel. D.B. and Cone, E.J.: Pharmacodynamic and pharmacokinetic action of ketocyclazocine 
enantiomers in the dog: Absence of sigma- or phencyclidine-like activity. J. Pharmacol. Exp. Ther. 

Casceila. N.G., Pearlson. G. Wong, D.F., Brousolle. E.P.M.. Nagoshi. C, Margolin, R.A. and London. 
E.D.: Effects of substance abuse on ventricular and sulcal measures assessed by computed tomography. 
British J. Psychiatry. 

Wu, X-.Z., Bell. J.A.. Spivak, C.E., London, E.D. and Su, T-.P.: Electrophysiological and binding studies 
on intact NCB-20 cells suggest presence of a low affinity sigma receptor. J. Pharmacol. Exp. Ther. 



ABSTRACTS 

McCann, D.J. and Su, T-.P.: Evidence that haloperidol inhibits the binding of (+)[%|SKF- 10047 to o 
sites through a competitive interaction. FASEB J. . 4:A329, 1990. 

Wu, X-.Z., Bell, J.A., Spivak, C.E., London. E.D. and Su, T-.P.: Sigma ligand [ 3 H]d-SKF- 10047 labelled 
two types of binding sites in intact NCB-20 cells. FASEB J. . 4:A329, 1990. 

McCann, D.J. and Su, T-.P.: Halopendol-sensitive (+) [ 3 H]SKF-10047 binding sites ("o sites") exhibit a 
unique distribution in rat brain subcellular fractions. Trans. Am. Soc. Neurochem . 21:247, 1990 



16 



Su, T-.P., Chien. S.F. and Oeltgen, P.R.: Hibernation induction trigger (HIT) extends preservation time 
in an autoperfusion muitiorgan preparation. Abstr. Am , fioc, CJi a, PharmacoL Tber .. 1990, In press. 

Morgan, M.J.. Cascella. N.G., Stapleton, J.M., Shaya, E.K., Wong, D.F. and London, E.D.: Relationship 
between ventricle/brain ratio, a measure of cerebral atrophy, and subjective responses to intravenous 
cocaine in human substance abusers. Soc. Neurosci. Abstr . 1£:582. 1990. 

Raymon. L.P., Kimes, A.S., Meltzer, L.T., Heffner, T.G. and London, E.D.: Effects of CI-943, A 
potential antipsychotic agent on cerebral glucose utilization in rats. Soc. Neurosci. Abstr . 16_:250, 1990. 

Vaupel, D.B., Della-Puppa, A., Lange, W.R. and London, E.D.: Verapamil reduces hypercapnia and 
euphoria produced by morphine in humans. Soc. Neurosci. Abstr. 16:928, 1990. 

Majewska, M.D., Bell, J. A. and London. E.D.: Redox phenomena modulate function of the NMDA 
receptor. Soc. Neurosci. Abstr . i£:464, 1990. 

Demirgoren. S.D., Majewska. M.D. and London, E.D.: Dehydroepiandrosterone sulfate, a neurosteroid, 
binds to rat brain membranes and modulates the function of GABA-A receptors. Soc. Neurosci. Abstr . 
16:691, 1990. 

Mantione, C.R., Demirgoren, S. and London. E.D.: Polyamine binding sites on synaptosomal 
membranes. Soc. Neurosci. Abstr . 16.:540, 1990. 

Kimes, A.S.. Wong, D.F. and London, E.D.: Use of mecamylamine (MEC) and lobeline (LOB) to 
estimate nonspecific binding of radiolabeled nicotine (NIC) in vivo. Soc. Neurosci. Abstr . 1^:684,1990. 

Wong, D.F., Gibson, R., London. E.D., Burns, H.D., Shaya. E., Dannals. R.F, Ravert, H.T.. Wilson, 
A. A. and Wagner Jr., H.N.: In vivo studies of o receptors with radiolabeled haloperidol. Soc. Neurosci. 
Abstr . 16:1140, 1990. 

Zaczek, R.. Culp, S., McCann, D.J. and De Souza, E.D.: Sequestration of 3 H-amphetamine into rat brain 
synaptosomes. Trans. Am. Soc. Neurochem. 21:248. 1990. 

London, ED.: Acute effects of cocaine on cerebral glucose metabolism in substance abusers. Am. 
College of Neuro-Psvchopharmaco l of rY Ann ual Meeting . San Juan, PR, Dec. 9-14, 1990. 

London, E.D.: Sigma receptor deficits in schizophrenia: Postmortem studies suggest a potential for PET 
diagnosis. Am. College of Neuro-Psvchopharr niiH"frv A nn "al Meeting . San Juan, PR, Dec. 9-14, 1990. 



17 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00200-05 NPL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: 

CEREBRAL EFFECTS OF ABUSED DRUGS: BRAIN IMAGING IN HUMANS AND 
LABORATORY ANIMALS 

Principal Investigator: 

ED. London (Chief, Neuropharmacology Lab, NPL, ARC) 

Others from NPL. ARC: Kimes, A.S. (Biologist), Phillips, R.L. (Sr. Staff Fellow) 

Stapleton, J.M. (Staff Fellow ), Morgan, M J. (Visiting Fellow), Yung, C.-K. (Visiting Fellow ) 

Cooperating Units: 

ARC: Heming, R. (Visiting Scientist. Psychology of Vulnerability and Cognitive Studies Laboratory, 

VCS) 

The Johns Hopkins Medical Institutions: Wong, D.F., Links. J., Dannals, R.F., Grayson, R., Wagner, 

H.N., Jr. 

Lab/Branch: Neuropharmacology Laboratory/Neuroscience Branch 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 5.60 Professional: 4.10 Other :_L5 

Check Appropriate Boxes: 

_ Human Subjects _ Human Tissues _x Neither 

_ Minors 
_ Interviews 

Summary of Work: 

Human positron emission tomographic (PET) scanning, using the [i8F]fluoro-deoxyglucose (FDG) 
method in placebo-controlled crossover studies, showed that euphorigenic doses of morphine or cocaine 
decrease rCMRglc. especially in cortical regions. The findings suggest that drugs of abuse produce 
euphoria by a mechanism which involves a reduction of rCMRglc. This hypothesis is being tested by 
examining the effects of other abused drugs (nicotine, buprenorphine ). Future PET studies with FDG will 
be used to obtain an objective measure of the opioid abstinence syndrome in human subjects, in order to 
test the effects of potential medications. 

Effects of nicotine and psychomotor stimulants on rCMRglc were studied by the deoxyglucose (DG) 
method in rats. Whereas nicotine stimulated rCMRglc in a pattern reflecting the distribution of nicotinic 
receptors, chronic nicotine produced tolerance, seen as a reduced response in some areas. In Fischer- 344 
rats, cocaine increased rCMRglc in motor areas and reduced rCMRglc in the lateral habenuia. Lewis rats 
were more sensitive to the effects than Fischer- 344 rats, and showed reductions in rCMRglc of the cortex, 
suggesting that genetic differences may influence susceptibility to cocaine abuse. Studies in gallamine- 
treated rats demonstrated that effects of psychomotor stimulant drugs on rCMRglc in motor systems are 
not secondary to limb movement. 



18 



The DG method also was used to identify brain areas important to opioid analgesia and the opioid 
abstinence syndrome. Although the formalin pain model was not associated with significant differences 
in rCMRglc as compared with control, morphine produced a dose-dependent reduction of rCMRglc in 
thalamic nuclei as well as in other areas implicated in nociception. The DG method was used to show 
that hypermetabolism in the brain, precipitated by administration of naloxone to opioid-dependent rats, is 
reversed by doses of clomdine which attenuate the opioid abstinence syndrome. Thus, efficacy of 
therapeutic interventions may be monitored by in vivo brain imaging in humans, using PET. 

PUBLICATIONS 

London, E.D., Broussolle. E.P.M., Links, J.M., Wong, D.F., Cascella, N.G., Dannals, R.F., Sano, M., 
Heming, R., Snyder, F.R., Rippetoe, L.R., Toung, T.J.R., Jaffe, J.H. and Wagner, Jr., H.N.: Morphine- 
induced metabolic changes in human brain - studies with positron emission tomography and [fluorine 
181fluorodeoxyglucose. Arch. Gen. Psvchiat. 47:73-81. 1990. 

Kimes. A.S.. Bell, J.A. and London. E.D.: Qonidine antagonizes increased glucose metabolism during 
naloxone-precipitated morphine withdrawal. Neuroscience 34(3):633-644, 1990. 

London. E.D.. Wilkerson, G.W., Ori, C. and Kimes, A.S.: Central action of psychomotor stimulants on 
glucose utilization in extrapyramidal motor areas of the rat brain. Rr a'Hl Res. 512:155-158. 1990. 

Henningfield J.E., London. ED. and Benowitz, N.L.: Arterio-venous differences in plasma 
concentrations of nicotine alter cigarette smoking. JAMA 263 :2049-2050, 1990. 

London, E.D., Cascella, N.G., Wong, D.F., Phillips, R.L., Dannals, R.F, Links, J.M., Heming, R., 
Grayson. R.. Jaffe. J.H. and Wagner, Jr., H.N.: Cocaine-induced reduction of glucose utilization in 
human brain: A study using positron emission tomography and [fluorine 18]fluorodeoxyglucose. Arch. 
Gen. Psvchiat . 4J:567-574, 1990. 

London. E.D.: Effects of nicotine on cerebral metabolism. In: The Biology of Nicotine Dependence 
(Ciba Foundn. Symp. No. 152), (G Bock and J. Marsh, eds.) Wiley, Chichester, pp. 131-146, 1990. 

London, E.D., Fanelli, R.J., Kimes. A.S. and Moses. R.L.: Effects of chronic nicotine on cerebral 
glucose utilization in the rat. Brain Re <; 5_2Q:208-214, 1990. 

Phillips, R.L.. London, E.D., Links, J.M. and Cascella, N.G.: Program for PET image alignment: 
Effects on calculated differences in cerebral metabolic rates for glucose. J. Nucl. Med 3.1:2052-2057, 
1990. 

London, E.D.: Glucose metabolism: An index of nicotine action in the brain. In: Proceedings of 
International Symposium on Nicotine. June 28-30, 1990, Hamburg. Advances in Pharmacological 
Sciences . (A. Adlkofer, Ed.), Birkhauser Verlag, Basel. 1990. 

London, E.D., Morgan, M.J., Phillips, R.L.. Stapleton, J.M., Cascella, N.G. and Wong, D.F: Mapping 
the metabolic correlates of drug-induced euphoria. NTDA Rese arch Monograph: Proceedings of the 
committee on Problems of Drug Dependence 1990. 

Cohen, S.R., Kimes, A.S. and London, E.D.: Morphine decreases cerebral glucose utilization in limbic 
and forebrain regions while pain has no effect. Neuropharmacology, in press. 

Cascella, N.G., Pearlson, G., Wong, D.F., Brousolle, E.P.M., Nagoshi. C, Margolin, R.A. and London, 
E.D.: Effects of substance abuse on ventricular and sulcal measures assessed by computed tomography. 
British J. Psychiatry , in press. 



19 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00202-07 NPL 



Period Covered: January 1, 1990 to December 3 1 , 1990 

Title of Project: 

PHYSIOLOGICAL EFFECTS OF OPIOIDS 

Principal Investigator: 

E.D. London (Chief. Neuropharmacology Lab, NPL, ARC) 

Others from NPL, ARC: Su, T.-P. (Pharmacologist), Bell, J A. (Pharmacologist), 

Vaupel, D.B. (Pharmacologist), Wu, X.-Z. (Visiting Associate), 

Cooperating Units: 

ARC: Goldberg, S. (Chief, Preclinical Parmacology Branch), Katz, J. (Chief, Psycholobiology Lab), 
Cone, E. (Chief, Chemistry & Drug Metabolism Lab.) 

Lab/Branch: Neuropharmacology Laboratory/Neuroscience Branch 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.10 Professional: 2.10 Other :_0JHI 

Check Appropriate Boxes: 

_x Human Subjects Human Tissues Neither 

Minors 

_ Interviews 

Summary of Work: 

We initiated studies on the interactions of verapamil (V), a Ca2+ channel antagonist, with morphine (M) 
in human volunteers. Preliminary findings suggest the following conclusions: 1. V reduces euphorigenic 
effects of M; 2. V reduces M-induced respiratory depression; and 3. V does not affect opioid analgesia. 

Using the isolated spinal cord, nifedipine, another Ca2+ channel blocker, and opioids block long latency - 
fiber reflexes, but have no effect on short latency monosynaptic reflexes. In contrast, in dorsal root 
ganglion cell cultures, opioids block N-type Ca2+ channels and nifedipine blocks L-type channels. Thus, 
Ca2+ channel antagonists and opioids both interfere with synaptic transmission, but they appear to act at 
different Ca2+ channels within the neuronal membrane. 

Comparisons of LAAM. norLAAM and dinorLAAM with morphine and methadone in the dog are being 
analyzed using a new parallel line bioassay program. NorLAAM is the most potent with respect to acute 
effects and in suppressing morphine withdrawal. In addition, LAAM and both metabolites produce acute 
physical dependence. 

Binding studies suggest that opioid receptors may be involved in behavioral conditioning. Rats trained 
under an fixed-ratio schedule became supersensitive to the rate-suppressing effects of naltrexone after 
weekly injections of naltrexone. Both kappa (k) and delta (5), but not mu (p) receptors were altered in 
certain brain areas suggesting that conditioning may involve changes in k and 8 opioid receptors. 



20 



In a multiple organ perfusion system, hibernation induction trigger (HIT) and DADLE, a synthetic 8 
opioid peptide, prolonged organ survival time. Mechanisms of actions of HIT and DADLE in this 
preservation model are under investigation. 

The pharmacology of the pentapeptide BW942C was examined in human and animal urination studies, 
isolated tissue preparations and binding studies. Results of dose-response analysis, antagonism studies 
and tolerance-cross tolerance paradigms suggest that BW942C is the first identified partial k agonist that 
is also a full u. agonist. 



PUBLICATIONS 

Vaupel. D.B , Cone, E.J., Johnson. R.E. and Su, T-.P.: Kappa opioid partial agonist activity of the 
enkephalin-like pentapeptide BW942C based on urination and in vitro studies in humans and animals. J_j 
Pharmacol. Exp. Ther . 252:225-234. 1990. 

Schindler. C.W., Wu. X-.Z.. Su, T-.P.. Goldberg, S.R. and Katz, J.L.: Enhanced sensitivity to the 
behavioral effects of naltrexone in rats: A conditioning phenomenon associated with opioid receptor 
changes. J. Pharmacol. Exp. Ther. 252 :8-14. 1990. 

Su. T-.P.. Chien, S.F. and Oeltgen, PR.: Hibernation Induction Trigger (HIT) extends preservation time 
in an autoperfusion multiorgan preparation. Clin. Pharmacol. Ther. 47:140. 1990. 

Bell. J. A.: Naloxone-induced facilitation of C-fiber reflexes is reduced by chronic morphine. Eur. J. 
Pharmacol. 168 :101-105. 1990. 

Vaupel, D.B. and Cone. E.J.: Pharmacodynamic and pharmacokinetic action of ketocyclazocine 
enantiomers in the dog: Absence of sigma- or phencyclidine-like activity. J. Pharmacol. Exp. Ther. . in 
press. 



21 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00312-01 NPL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: LIGAND-GATED ION CHANNELS 

Principal Investigator: 

E.D. London (Chief, Neuropharmacology Lab, NPL, ARC) 

Others From NPL, ARC: Mantione, C.R. (Sr. Staff Fellow,), Majewska, M.D. (Sr. Staff Fellow), Spivak, 

C.E. (Pharmacologist) 

Cooperating Units: 

Lab/Branch: Neuropharmacology Laboratory/Neuroscience Branch 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 5.20 Professional: 4.62 Other:_,4Jl 

Check Appropriate Boxes: 

_ Human Subjects Human Tissues _x Neither 

Minors 

_ Interviews 

Summary of Work: 

This project focuses on the receptor superfamily of ligand-gated ion channels, including the N-methyl-D- 
aspartate receptor (NMDA-R), the nicotinic cholinergic receptor (nAChR), and the GABAa receptor 
(GABA-R). 

NMDA-R is of interest because phencyclidine binds to sites inside the open state of the ionic channel 
formed by the receptor. This receptor has been linked to neurodegeneration in various conditions, 
including ischemia, epilepsy, and Huntington's disease. Our studies show that NMDA-R is regulated by 
tissue redox phenomena, as some reducing agents (e.g., ascorbic acid, hydroquinone) produce 
inactivation, whereas others (dithiothreitol) facilitate receptor activation. Furthermore, we found that 
ascorbic acid protects cultured cortical cells from the NMDA- and glutamate-induced toxicity. Using 
pHJspermidine, we have identified and characterized polyamine receptors, which also appear to 
modulate activity of NMDA-R. Our assay system may provide a template for the design of therapeutic 
agents to alter the function of the NMDA-R. 

[3H]Mecamylamine was used as a new radioligand probe for the canonic channels of nAChRs. We 
discovered that the binding of [3H]mecamylamine and [3H]chlorpromazine is markedly increased by 
some purinergic nucleotides (e.g., ATP), suggesting that they may be important modulators of nAChR. 
Preliminary studies have shown that polyamines negatively modulate the binding of [3H]mecamylamine 
but not [3H]methylcarbamylcholine (a ligand for the acetylcholine recognition site of nAChRs) in brain. 
Since both receptors are ligand-gated canonic channels, it seems possible that polyamines generally are 
regulators of such receptors. 

Pregnenolone sulfate (PS) and dehydroepiandrosterone sulfate (DHEAS) are modulators of GABA-R. 
We examined binding of [3H]PS in the rat brain, noting that this steroid seems to interact at the interface 



22 



of the receptor with associated phospholipids. We identified and characterized specific binding sites for 
[3H1DHEAS. and demonstrated, by biochemical and electrophysiological methods, that DHEAS is a 
noncompetitive negative modulator of the GABAa receptor complex. 

PUBLICATIONS 

Mantione. C.R., Demirgoren, S. and London. E.D.: Specific binding of [ 3 H]spermidine to membranes of 
rat brain. Eur. J. Pharmacol. l£0.:393-394, 1990. 

Majewska, M.D., Demirgoren. S.. Spivak, C.E. and London. E.D.: The neurosteroid 
dehydroepiandrosterone sulfate is an allosteric antagonist of the GABA^ receptor. Brajfl RfiS, 5_26.:143- 

146. 1990. 

Majewska, M.D.: Steroid regulation of GABA receptor: ligand binding, chloride transport, behavior. In: 
Steroids and Brain Activity (Ciba Foundn. Symp. No. 153), (G. Bock and J. Marsh, eds.) Wiley, 
Chichester. 1990. PP. 83-106. 

Majewska, M.D.. Demirgoren, S. and London. E.D.: Binding of pregnenolone sulfate to rat brain 
membranes suggests multiple sites of steroid action at the GABA^ receptor. Eur. J. Pharmacol. . Mol. 

Pharmacol. Section 189:307-315. 1990. 

Majewska. M.D.: Neurosteroids, metabolism and function. Neuroscience Facts 1:2, 1990. 

Majewska, M.D., Bell, J.A. and London, E.D.: Regulation of the NMDA receptor by redox phenomena: 
Inhibitory role of ascorbate. Brain Res. 537:328-332, 1990 

Majewska. M.D. and Bell, J.A.: Ascorbic acid protects neurons from injury induced by glutamate and 
NMDA. Neuxoreport 1:194-196, 1990. 

London, E.D., Dawson, V.L. and Mantione. C.R.: Specific binding sites for polyamines in mammalian 
brain. In: NMDA Related Agents: Biochemistry. Pharmacology and Behavior. Proceedings from a 
satellite symposium of the 17th Congress of Collegium Internationale Neuro-Psychopharmacologicum. 
Nagoya, Japan. Sept. 16-17. 1990. NPP Books, Inc., Ann Arbor. MI. T. Nabeshima, T. Kameyama. and 
E.F. Domino, eds. 

Spivak. C.E., Waters. J.A and Aronstam, R.S.: (+)-C)ctahydro-2-methyl-{rari5-5(lH)-isoquinolone 
methiodide: A probe to test the steric limits of agonists active at the nicotinic acetylcholine receptor. L 
Mol. Graphics , in press. 



23 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00206-06 NPL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: SIGMA RECEPTORS 
Principal Investigator: 

Su, T.-P. (Pharmacologist) 

Others from NPL, ARC: London, E.D. (Chief, Neuropharmacology Laboratory), Spivak. C.E. 
(Pharmacologist), Bell, JA. (Pharmacologist), Wu, X.-Z. (Visiting Associate), McCann, D. (Staff 
Fellow), Kimes, A.S. (Biologist) 

Cooperating Units: 

Lab/Branch: Neuropharmacology Laboratory/Neuroscience Branch 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: 2.53 Professional: 2.43 Other: 0.10 

Check Appropriate Boxes: 

_ Human Subjects _x Human Tissues Neither 

Minors 

_ Interviews 

Summary of Work: 

This project examines molecular, electrophysiological, and in vivo interactions of ligands for sigma (a) 
receptors. A unifying hypothesis for binding of o ligands. including steroids, "atypical" antipsychotics, 
such as BMY- 14802 and remoxipride. and the most selective o ligand. PRE-084. was formulated using 
conformational fitting and electrostatic potential calculations. All a ligands have a pharmacophore with 
three sites of interaction; the distances between the sites are identical for the ligands. Surface maps of 
electrostatic charges also are similar for all o ligands tested. 

Solubilized a receptors were labeled with (3H]progesterone, providing the first direct demonstration of 
steroid binding to the receptors. Solubilized o receptors also are modulated by certain anticonvulsant 
drugs. A preliminary estimation of the molecular weight of solubilized a receptors was obtained using 
molecular sizing chromatography. 

A low affinity o receptor that modulates the closing of a tonic potassium channel was demonstrated in 
the NCB-20 cells. The low affinity receptor may play an important physiological and pharmacological 
role in locomotion regulation. 

Studies of postmortem brains from schizophrenic patients demonstrated selective losses of o receptors in 
the temporal cortex and dentate nucleus of the cerebellum, suggesting a role of the o receptor in 
psychosis, and underscoring the importance of imaging the o receptor in vivo. Studies in mice indicated, 
that radiolabeled d-N-allylnormetazocine and haloperidol show potential to be developed as in vivo 
ligands. Studies of the binding of [l25r)p-iodophenyl amanantylguanidine (PIPAG), a potential SPECT 
ligand, to guinea pig brain showed high affinity (0.6 nM) and a pharmacological profile and 
neuroanatomical distribution typical of the classical a receptor. 



24 



Future work includes molecular modeling studies on other a ligands, further chemical and 
pharmacological studies of PRE-084, purification of o receptors, and biochemical studies on the 
mechanism of modulation of the low affinity o receptor on the tonic potassium channel. 



PUBLICATIONS 

Vu, T.H., Weissman, A.D. and London. E.D.: Pharmacological characteristics and distributions of o and 
pbencyclidine binding sites in the animal kingdom. J. Neurochem. 54:598-604, 1990. 

Su, T-.P., and Wu, X-.Z.: Guinea-pig vas deferens contains o but not phencyclidine receptors. Neurosci. 
LeJL 101:341-345, 1990. 

McCann. D.J. and Su, T-.P.: Haloperidol-sensihve (+)[ 3 H]SKF-10,047 binding sites ("a sites") exhibit a 
unique distribution in rat brain subcellular fractions. Eur. J. Pharmacol . 188 :211-218, 1990. 

McCann, D.J. and Su. T-.P.: Halopendol competitively inhibits the binding of (+)[%]SKF- 10.047 to o 
sites. Eur. J. Pharmacol . 1811:361-364, 1990. 

Weissman. E.D., Broussolle, E.P. and London, E.D.: In vivo binding of pjfld-N-allylnorrnetazocine and 
[-H]halopendol to sigma receptors in the mouse brain. J. Chem. Neuroanatomy 1:347-354, 1990. 

London. ED.: Studies of o receptors and metabolic responses to o ligands in the brain. In: Sigma. PCP 
and NMDA Receptor Systems . EB De Souza, ED London, and D Clouet, eds. NIDA Research 
Monographs. 

Gund. T.M.. Shukla. K. and Su. T-.P.: A unifying hypothesis for binding of psychotomimetic ligands at 
the sigma receptor. Chem. Pes. Autom. News 5_:l-20, 1990. 

Su. T-.P.: Pharmacological characterizations of a receptors. In: Sigma. PCP and NMDA Receptor 
Systems . NIDA Research Monographs. EB De Souza. ED London, and D Clouet. eds. DHHS. NIDA. 

Su, T-.P., Shuklar. K. and Gund, T.: Steroid binding at o receptors: CNS and immunological 
implications. In: Steroids and Neuronal Activity (Ciba Foundn. Symp. No. 153), (D.J. Chadwick, K. 
Widdows, eds.) Wiley, Chichester. 1990. 

Weissman. A.D., Cassanova, M.F., Kleinman, J.F., London, E.D. and De Souza, E.B.: Selective 
reduction of cerebral cortical sigma. but not PCP binding sites in schizophrenia. Biol. Psvch . in press. 

Su, T-.P., Wu, X-.Z., Spivak, C.E., London, E.D. and Bell, J.A.: Binding studies on intact NCB-20 cells 
suggest s receptor multiplicity: at and ai. In: NMDA Related Agents: Biochemistry. Pharmacology and 
Behavior . Proceedings from a satellite symposium of the 17th Congress of Collegium Internationale 
Neuro-Psychopharmacologicum. Nagoya, Japan, Sept. 16-17, 1990, NPP Books, Inc., Ann Arbor, MI, T. 
Nabeshima, T. Kameyama, and E.F. Domino, eds. 

Wu, X-.Z.. Bell, J.A., Spivak, C.W., London, E.D. and Su, T-.P.: Electrophysiological and binding 
studies on intact NCB-20 cells suggest presence of a low affinity sigma receptor. J. Pharmacol. Exp. 
Ther. . in press. 



25 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00210-05 NPL 



Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: HIV INFECTION AND DRUG ABUSE 
Principal Investigators: 

Kimes, A.S. (Biologist, Neuropharmacology Laboratory) 
Other London, E.D. (Chief, Neuropharmacology Laboratory) 

Cooperating Units: 

USAMRICD, Aberdeen Proving Ground, MD, Smith, W.J. (Research Chemist, Biochemical 

Pharmacology Branch), 

Intramural Research Program, NIAAA. Bethesda, MD, Tabakoff, B. (Director), 

Szabo, J. (Visiting Fellow) 

Lab/Branch: Neuropharmacology Laboratory/Neuroscience Branch 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.15 Professional: 0.15 Other: 0.00 

Check Appropriate Boxes: 

_ Human Subjects Human Tissues _x Neither 

Minors 

_ Interviews 

Summary of Work: 

Human immunodeficiency virus (HIV) infection in the central nervous system is characterized by 
dementia and deficits in motor functions. Cerebral metabolic effects of gpl20 (a HTV viral envelope 
glycoprotein), which binds to brain membranes and T-lymphocytes, were studied in rats. 
Intracerebroventricular injection of gpl20 reduced the regional cerebral metabolic rate for glucose in the 
lateral habenula and the suprachiasmatic nucleus, and produced an overall decrease in glucose 
metabolism. As reduced glucose metabolism is observed using positron emission tomography in humans 
presenting with HlV-associated dementia, the findings suggest that gp!20 can alter neuronal function and 
contribute to HTV-related dementia. 

As intravenous drug abusers show an abnormally high incidence of HTV infection, animal studies were 
performed to determine if chronic exposure to abused substances alters immune function. Mice receiving 
chronic opioid treatment had fewer circulating T-Iymphocytes (helpers and suppressor/cytotoxic) than 
concurrent controls. The effect was dose-dependent, was not blocked by treatment with the opioid 
antagonist naltrexone, and could be observed within 24 h of initiation of treatment. Oxymorphone 
caused a similar effect. Morphine treatment did not affect mitogen-stimulated lymphocyte proliferation. 
This work suggests that morphine compromises immunocompetency and that the use of opioids by 
intravenous drug abusers may increase the incidence of infection subsequent to exposure to bacterial and 
viral agents. 



26 



PUBLICATIONS 

Kimes, A.S., London, E.D., Szabo, G., Raymon, L. and Tabakoff. B.: Reduction of cerebral glucose 
utilization by the HIV envelope glycoprotein gp-120. Exp. Neurol. , in press. 



27 



3. Neurobiology Laboratory - Errol B. De Souza. Ph.D., Chief. 

Overview 

The Laboratory of Neurobiology conducts research on the neurobiological underpinnings of drug abuse 
and addiction. At present, the Laboratory has three major areas of research which include 1 ) the study of 
the neuroendocrine aspects of addiction, with a focus on stress, hypothalamic peptides and drugs of 
abuse, 2) the study of the pharmacological and neurotoxic effects of drugs of abuse, and 3) the study of 
the brain-neuroendocrine-immune axis and its related peptides, hormones, lymphokines and monokines. 
The Laboratory utilizes a multifaceted approach which includes biochemical, cellular, pharmacological, 
neuroendocrine and neuroanatomical techniques to investigate the problems outlined above. 

During the past year Dr. Errol B. De Souza has taken a position in industry and has left the ARC. At this 
point in time, a review of the Laboratory is being conducted and a search is made for a new Laboratory 
Chief. Within the next year, it is expected that plans will be made for the utilization of the resources in 
this laboratory. 

Summary of Ongoing Research 

Research has been conducted in a variety of areas related to drug abuse. One area has been corticotropin 
releasing factor (CRF) which is a stress-related neurotransmitter in the central nervous system. Relapse to 
addiction often is associated with stress and molecular mechanisms of stress have been explored in this 
context. The main focus of this program has been the CRF receptor and a number of studies have been 
carried out on this receptor. 

Another area of research involves the role of neurotransmitters and their receptors in human 
neuropsychiatric disorders and neurodegenerative diseases. For example, changes in sigma and PCP 
receptors have been examined in the brains from human populations. 

The designer drugs MDA and MDMA have potent, long-lasting neurotoxic effects in brain. These have 
been explored in rodent models of toxicity. Both biochemical and neurohistological changes have been 
identified. 

Additional projects include an examination of neurochemical, neuroendocrine, and neurotoxic effects of 
other drugs. Also, interactions of the brain-endocrine-immune axis have been examined. In particular, 
receptors for interleukin- 1 have been examined. 



28 



Publications 

Grigoriadis. D.E., Zaczek, R., Peaisall. D. and De Souza. Solubilization of high-affinity corticotropin- 
releasing factor (CHI 7 ) receptors from rat brain: Characterization of an active digitonin-solubilized 
receptor complex. Endocrinology 125:3068-3077, 1989. 

Zaczek, R.. Culp, S.G. and De Souza, E.B. Intrasynaptosomal sequestration of PH]-methylene- 
dioxyamphetamine: Characterization suggests the presence of a factor responsible for maintaining 
sequestration. J. Neorocbem. 54:195-204. 1990. 

Appel, N.M.. Mitchell, Wm.M., Contrera, J.F. and De Souza, E.B. Effects of high-dose fenfluramine 
treatment on monoamine uptake sites in rat brain: Assessment using quantitative autoradiography. 
Svnapse 6:33-44. 1990. 

Zaczek. R.. Battaglia, G.. Culp, S., Appel, N.M.. Contrera, J.F. and De Souza, E.B. Effects of repeated 
fenfluramine administration on indices of monoamine function in rat brain: Pharmacokinetics, dose 
response, regional specificity and time course data. J. Pharmacol. Exp. Ther. 253:104-1 12, 1990. 

Zaczek. R.. Fritschy. J-M. Culp S., De Souza, E.B. and Grazanna R. Differential effects of DSP-4 on 
norepinephrine uptake into synaptosomes from cerebral cortex and hypothalamus: Evidence for 
heterogeneity of the norepinephrine uptake sites. Brain Research 522:308-314, 1990. 

Takao, T.. Mitchell. Wm.M., Tracey, D.E. and De Souza, E.B. Identification of interleukin- 1 receptors 
in mouse testis. Endocrinology 127:251-258, 1990. 

Webster. E.L.. Tracey, D.E., Jutila. M.A., Wolfe, S.A. Jr. and De Souza, E.B. Corticotropin-releasing 
factor (CRF) receptors in mouse spleen: Identification of receptor bearing cells as resident macrophages. 
Endocrinology 127:440-452, 1990. 

Bitar, M.S. and De Souza. E.B. Diabetes-related changes in brain beta adrenoreceptors in rats as assessed 
by quantitative autoradiography: Relationship to hypothalamic norepinephrine metabolism and pituitary- 
gonadal hormone secretion. J. Pharmacol. Exp. Ther. 254:781-785. 1990. 

Takao. T.. Tracey, D.E.. Mitchell. Wm.M. and De Souza. E.B. Interleukin-1 receptors in mouse brain: 
characterization and neuronal localization. Endocrinology 127:3070-3078, 1990. 

Appel. N.M.. Mitchell. Wm.M., Garlick. R.K., Glennon. R.A., Titeler, M. and De Souza, E.B. 
Autoradiographic characterization of [I25JDOI: A novel phenyhsopropylamine derivative which labels 
both 5HT2 and 5HT| C receptors. J. Pharmacol. Exp. Ther. 225:843-857, 1990. 

McLeod. D.R.. Hoehn-Saric, R., Zimmerli, W.D., De Souza, E.B. and Oliver, L.K. Treatment effects of 
Alprozolam and Imipramine: Physiological versus subjective changes in patients with generalized 
anxiety disorder. Biological Psychiatry 28:849-861, 1990. 

Weissman. A.D., Casanova, M.F., Kleinman, J.E., London. ED. and De Souza, E.B. Selective reduction 
in cerebral cortical sigma . but not PCP binding sites in schizophrenia. Biological Psychiatry 29:41.54, 
1991. 

Weissman, A.D., Casanova, M.F., Kleinman, J.E. and De Souza, E.B. PCP and sjgma. receptors in brain 
are not altered after repeated exposure to PCP in man. Neuropsvcbopharmacologv 4:95102, 1991. 

Yeh, S.Y and De Souza, E.B. Lack of neurochemical evidence for neurotoxic effects of repeated cocaine 
administration in rats on brain monoamine neurons. Drug and Alcohol Dependence 27:51-61, 1991. 



29 



Battaglia, G., Zaczek. R. and De Souza, E.B. MDMA effects in brain: Pharmacologic profile and 
evidence of neurotoxicity from neurochemical and autoradiographic studies. In: MDMA: "Ecstasy" 
nnnVor Human Neurotoxin? (S.J. Peroutka, ed ) Kluwer Academic Publishers. Boston, pp. 171-199, 1989. 

De Souza, ED. Autoradiographic localization of monoamine and corticotropin-releasing factor (CRF) 
receptors in the pituitary: effects of glucocorticoids and peripheral amines. In: Stress. Neurochemical 
and Humoral Mec h a nisms (G.R. Van Loon, R. Kvetnansky, R. McCarty and J. Axelrod, eds.), Gordon 
and Breach Science Publishers. New York, pp. 391-407, 1989. 

De Souza. E.D., Webster, E.L. Grigoriadis, D.E. and Tracey, D.E. Corticotropin-releasing factor (CRF) 
and Interleukin- 1 (IL-1) receptors in the brain-pituitary-immune axis. Psychoph annacology Bulletin 
25:299-305, 1989. 

De Souza, E.B. and Insel. T.R. Corticotropin-releasing factor (CRF) receptors in the rat central nervous 
system: autoradiographic localization studies. In: Corticotropin-Releasing Factor: Basic and Clinical 
Studies of a Neuropeptide . (E.B. De Souza and C.B. Nemeroff. eds.), CRC Press, Boca Raton, FL., pp. 
69-90, 1990. 

De Souza, E.B. and Grigoriadis, D.E. Corticotropin-releasing factor (CRF) receptors in brain: 
characterization and regulation. In: Corticotropin-Releasing Factor Basic a "d. Clinic al Studies of a 
Neuropeptide . (E.B. De Souza and C.B. Nemeroff. eds.), CRC Press. Boca Raton. FL.. pp. 115-135, 
1990. 

Battaglia, G., Webster, E.L. and De Souza, E.B. Characterization of second messengers coupled to 
corticotropin-releasing factor (CRF) receptors in brain. In: Corticotropin-Releasing Factor: Basic and 
Clinical Studies of a Neuropeptide . (E.B. De Souza and C.B. Nemeroff, eds.), CRC Press, Boca Raton, 

FL., pp. 335-349, 1990. 

De Souza. E.B. and Appel, N.M. Distribution of brain and pituatary receptors involved in mediating 
stress responses. In: Neurobiology and Neuroendocrinologv of Stress (M.R. Brown, C. Rivier and GF. 
Koob, eds.), Marcel Dekker, Inc., New York, pp. 91-117, 1990. 

De Souza, E.B. Role of corticotropin-releasing factor in neuropsychiatric disorders and 
neurodegenerative diseases. Annu al Reports in Medicinal Chemistry . Vol. 25 (Topics in Biology): 215- 
224, 1990. 

De Souza, E.B. Neurotransmitter Receptor Imaging Techniques. In: Neuropepti des in Psychiatry 
(Progress in Psychiatry Series) (C.B. Nemeroff, ed) 29:261-278, 1990. 

De Souza, E.B. Neuroendocrine effects of benzodiazepines. L Psychiatric Research 24(52): 11 1-1 19, 
1990. 

Appel, N.M., Zaczek, R., Mitchell, Wm.M. and De Souza, E.B. Immunohistochemical and auto- 
radiographic investigations of high dose fenfluramine treatment on monoamine neurons in rat brain. In: 
Proceedings of Int. Svmp. on Serotonin: From Cell Biology to Pharmacology and. Therapeutics (P. 
Paoletti. P.M. Vanhoutte, N. Brunello and F.M. Maggi, eds.) Kluwer Academic Publishers, Boston pp. 
625-629, 1990. 

Titeler, M., Appel, N.M., De Souza, E.B. and Glennon, R.A. Receptor pharmacology of MDMA and 
related hallucinogens. Annals New York Acad. Sci. (The Neuropharmacology of Serotonin, P.M. 
Whitaker-Azmitia and S.J. Peroutka, eds.) Vol. 600:626-629, 1990. 



30 



De Souza. E.B., Battaglia. G. and Insel, T.R. Neurotoxic effects of MDMA on brain serotonin neurons: 
Evidence from neurochemical and radioligand binding studies. Appals N ew Yoik A cad. Sci. (The 
Neuropharmacology of Serotonin), Vol. 600:682-698, 1990. 

De Souza. E.B., Takao, T. and Tracey, D.E. Interieukin-1 receptors in the brain-endocrine-immune axis. 
Proceedings of the 17th C.I.N.P. Congress, Clinical Pharmacology, 1990. 

De Souza, E.B. Psychomimetic and neurotonic effects of amphetamines and related designer drugs. In: 
Practical Clinical Management: Drug Abuse Education for the Primary Care Physician. Medical and 
Chirurgical Faculty of Maryland , pp. 79-84, 1990. 

Webster, E.L., Grigoriadis. D.E. and De Souza. E.B. Corticotropin- releasing factor receptors in the 
brain-pituitary-immune axis. In: Stress. Neuropeptides, and Systemic Disease (J.A McCubbin, P.G. 
Kaufmann and C.B. Nemeroff. eds.), Academic Press, Inc., San Diego, pp. 233-260, 1991. 

Articles in Press 

Goeders. N.E., Bienvenu. O.J. and De Souza, E.B. Chronic cocaine administration alters corticotropin- 
releasing factor receptors in the rat brain. Brain Research (in press ). 

Blake. M.J.. Appel, N.M., Joseph. J.A.. Stagg, C.A., Anson, M., De Souza, E.B. and Roth. G.S. 
Muscarinic acetylcholine receptor subtype nRNA expression and ligand binding in the aged rat brain. 
Neurobiol. of Aging (in press). 

Zaczek, R.. Culp, S.. Goldberg, H., McCann, D.J. and De Souza, E.B. Interactions of [ 3 H]-amphetamine 
with rat brain synaptosomes: Part I. Saturable sequestration. J. Pharmacol. Exp. Ther. (in press). 

Zaczek. R., Culp, S. and De Souza, E.B. Interactions of [ 3 H]-amphetamine with rat brain synaptosomes: 
Part II. Active transport. J. Pharmacol. Exp. Ther. (in press). 

Appel, N.M.. Owens, M.J., Culp, S., Zaczek, R., Contrera, J.F., Bissette, G, Nemeroff, C.B. and De 
Souza, E.B. Role for brain corticotropin-releasing factor in the weight-reducing effects of chronic 
fenfluramine treatment in rats. Endocrinology (in press). 

Heroux, J.A., Grigoriadis. D.E. and De Souza, E.B. Age-related decreases in corticotropin-releasing 
factor receptors in the brain and anterior pituitary gland of the rat. Brain Research 542:155-158, 1991. 

Takao, T., Mitchell, Wm.M. and De Souza, E.B. Interieukin-1 receptors in mouse kidney: 
identification, localization and modulation by lipopolysaccaride treatment. Endorcrinology (in press). 

Battaglia, G., Sharkey, J., Kuhar, M.J. and De Souza. E.B. Neuroanatomic specificity and time course of 
alterations in rat brain sertonergic pathways induced by MNDA (3,4-methylenediozymethamphetamine): 
assessment using quantitative autoradiography. Svnapse (in press) 

Cunningham. E.T., Jr., Wada, E., Carter, D.B., Tracey, D.E., Battey, J.F. and De Souza, E.B. 
Localization of interieukin-1 receptor messenger RNA in murine hippacampus. Endocrinology (in press) 

Kuhar. M.J. and De Souza, E.B. Receptor autoradiography as an aid in explaining drug action. In: 
Im aging the functional neuroanatomy of drug action (E.D. London, ed.). The Telford Press, Caldwell, 
N.J. (in press). 



31 



De Souza, E.B., Grigoriadis. D.E. and Webster. E.L. Role of brain, pituitary and spleen corticotropin- 
releasing factor (CRF) receptors in the stress response. In: The Stress of Life. Revisited (Methods and 
Achievements in Experimental Pathology) (G. Jasmin and M. Cantin. eds.), S. Karger, Switzerland (in 
press*. 

Wolfe, S.A., Jr. and De Souza. E.B. Sigma receptors in the brain-endocrine-immune axis. In: Sigma. 
PCP and NMDA Receptor Systems (E.B. De Souza. E.D. London and D.H. Clouet. eds.), NIDA Research 
Monographs (in press). 

Grigoriadis, D.E. and De Souza. E.B. Biochemical, pharmacological and autoradiography methods to 
study corticotropin-releasing factor (CRF) receptors. Methods in Neuroscience . Vol. 5:510-538, Peptide 
Technology (P.M. Conn, ed.) Academic Press. Inc., Florida. 1991. 

De Souza. E.B. Corticotropin-Releasing Hormone receptors. In: Handbook of Chemical Neuroanatomy: 
Neuropeptide Receptors in the CNS. Part III (A. Bjorklund, T. Hokfelt and M.J. Kuhar, eds), Elsevier, 
Amsterdam (in press). 

Grigoriadis. D.E. and De Souza. E.B. Receptor binding techniques. In: Comprehensive Textbook of 
Neuroendorcrinologv (C.B. Nemeroff, ed. ), The Telford Press, Caldwell, N.J. (in press). 

Appel, N.M. and De Souza. E.B. Autoradiographic localization of non-receptor proteins in brain. In: 
Using Autoradiography and Correlative Imag in g in Vitro and In Vivo (W. Stumpf and H. Solomon, eds). 
Academic Press, Inc.. Florida. 

Weissman. AD. and De Souza, E.B. Postmortem investigations of Sigma and PCP receptors in 
psychosis. In: Exerpta Medica International Congress Series (5th World Congress of Biological 
Psychiatry, Florence) 1991. 

Abstracts Published 

Appel, N.M., Zaczek. R., Owens, M., Culp, S. Nemeroff. C.B. and De Souza, E.B. Relationships 
between brain serotonin (5HT) and corticotropin- releasing hormone (CRH) in the anti-obesity effects of 
fenfluramine. 1990 Winter Neuropeptide Conference, Breckenridge, Colorado, 1990. 

De Souza, E.B. Role of corticotropin-releasing factor (CRF) in the development of obesity syndromes 
and in the effects of antiobesity drugs. Winter Neuropeptide Conference, Breckenridge, Colorado, 1990. 

De Souza, E.B. The role of corticotropin-releasing factor (CRF and its receptors in coordinating the 
endocrine, autonomic and behavioral responses to stress. European Winter Conference on Brain 
Research, Les Arcs 2000, France. 

Zaczek, R., Culp, S., McCann, D. and De Souza, E.B. Sequestration of ^H-amphetamine into rat brain 
synaptosomes. American Society of Neurochemistry Meeting, Abstr. #324, 1990. 

Appel, N.M., Mitchell. Wm.M., Gariick, R.K.. Glennon, R.A., Titeler, M. and De Souza, E.B. 
Autoradiographic characterization of l25Mabeled 2,5-dimethoxy-4-iodophenylisopropylamine (DOI): A 
phenylisopropylamine derivative lableing both 5HT2 and 5HTi c receptors. FASEB J. . 4:A328, 1990. 

Wolfe, S.A., Jr., Aguayo, L.G. and De Souza, E.B. Sigma receptors in rat pineal gland: 
Electrophysiology and autoradiographic localization. FASEB J. . 4:A329. 1990. 



32 



Webster. E.L.. Tracey, D.E. and De Souza, E.B. Corticotropin-releasing factor (CRF) treatment 
upregulates interleukin- 1 (IL-1) receptors in AtT-20 putuitary tumor ceils. The Endocrine Society, 72nd 
Annual Meeting, Abstr. #379. 1990. 

Takao, T., Mitchell, Wm.M, Tracey, D.E. and De Souza, E.B. Identification of Interleukin- 1 receptors 
in mouse testis. The Endocrine Society, 72nd Annual Meeting, Abstr. #420, 1990. 

De Souza, E.B.. Takao, T. and Tracey, D.E. Interleukin-1 receptors in mouse brain. 2nd International 
Congress of Neuroendocrinology, Neuroendocrinology Vol. 52(S1):76, 1990. 

Heroux, J.A.. Grigoriadis. D.E. and De Souza, E.B. Age-related decreases in corticotropin-releasing 
factor receptors in the brain and anterior pituitary gland of the rat. Soc. for Neurosci - 16: 147, 1 990. 

Grigoriadis. D.E., Heroux, J. A., Pearsall, D.M. and De Souza, E.B. Biochemical isolation, 
characterization and partial purification of corticotropin-releasing factor (CRF) receptors from rat brain. 
Soc. for Neurosci .. 16:147, 1990. 

Zaczek. R.. Culp, S. and De Souza. E.B. High-affinity active transport of [^H]-d-amphetamine into rat 
striatal synaptosomes. Soc. for Neurosci . 16:303. 1990. 

Sharpe. L.G.. Pilotte. N.S., Mitchell. Wm.M.. De Souza, E.B. and Dax. E.M. Withdrawal from chronic 
cocaine decreased dopamine transporter sites in the rat nucleus accumbens (NAc). Soc. for Neurosci .. 
16:696. 1990. 

Wolfe, S.A. Jr., Aguayo, L.G. and De Souza. E.B. Sigma receptors in immune and endocrine tissues: 
Dichotomy between binding and function. Soc. for Neurosci . 16:801, 1990. 

Culp, S., Zazcek, R., Appel, N.M.. Contrera, J.F. and De Souza. E.B. Comparison of the effects of 
repeated oral versus subcutaneous d,l -fenfluramine administration on brain serotonin neurons in the rat. 
Soc. for Neurosci . 16:1033, 1990. 

Wada. E., Cunningham. E.T. Jr., Mitchell, Wm.M., Carter, D.B., Tracey, D.E.. Battey, J.F. and De 
Souza. E.B. Identification of interleukin-1 receptor mRNA in murine hippocampus. Soc. for Neurosci . 
16:1213, 1990. 

Takao, T., Tracey, D.E., Mitchell, Wm.M. and De Souza. E.B. Interleukin-1 receptors in mouse brain: 
Characterization and autoradiographic localization. Soc. for Neurosci . 16:1213, 1990. 

Appel, N.M.. Seamon. K.B., Laurenza, A., Simpson, I.A. and De Souza, E.B. Localization of adenylate 
cyclase and glucose transporter in rat brain using [I25l]-Labeled derivatives of forskolin. Soc. for 
Nsmosci 16:1302, 1990. 

Weissman, AD. and De Souza, E.B. Brain sipna and dopamine receptors are not modulated by chronic 
d-pentazocine administration in rats. Soc. for Neurosci . 16:1305, 1990. 

De Souza, E.B., Takao, T and Tracey, D.E. Interleukin-1 receptors in the brain-endocrine-immune axis. 
American College of Neuropsychopharmacology. San Juan. Puerto Rico, p. 66, 1990. 

De Souza, E.B. and Weissman, A.D. Neurotoxicity of phencyclidine and related drugs: Introduction to 
the problem and human autopsy data. American College of Neuropsychopharmacology, San Juan, Puerto 
Rico. p. 82, 1990. 



33 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00302-03 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Neurotoxic Effects of MDA and MDMA (Ecstasy) 
Principal Investigators: Cooperating Units 

P.I. E.B. De Souza Chief, Neurobiology Laboratory 

Others: R. Zaczek Staff Fellow, ARC 

N.M. Appel Staff Fellow, ARC 

A. Weissman Staff Fellow. ARC 

S.Y. Yeh Scientist, ARC 

T. Insel Scientist. LCS. NIMH 

Cooperating Unit: Laboratory of Clinical Science. NIMH 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3 00 Professional: 2.2 Other: 0.8 

Check Appropriate Boxes: 

Human Subjects Human Tissues X Neither 

Minors 

Interviews 

Summary of Work 

Substantial progress has been made in examining the mechanism of action and the effects ot 
amphetamine as well as amphetamine analogs. We have found that 3H-amphetamine is sequestered by 
synaptosomal fractions from brain. The neurotoxic effects of MDMA on brain serotonin neurons has 
been explored further by both biochemical techniques as well as autoradiographical techniques. Overall, 
these studies elucidate the mechanism of action of amphetamine as well as further explain the neurotoxic 
effects of drugs like MDMA. 

This project will not be continued in 1991. 



34 



PUBLICATIONS 

Zaczek. R., Culp, S.G. and De Souza. E.B. Intrasynaptosotnal sequestration of [ 3 H]-ampbetamine and 

[ 3 H]-methylenedioxy-amphetamine: Characterization suggests the presence of a factor responsible for 
maintaining sequestration. J. Neurochem , 54:195-204. 1990. 

Zaczek, R., Culp, S., Goldberg, H., McCann, D.J. and De Souza, E.B. Interactions of [ 3 H]-amphetamine 
with rat brain synaptosomes: Part L Saturable sequestration. J. Pharmacol. Exp. Ther. tin press). 

Zaczek. R., Culp, S. and De Souza, E.B. Interactions of [ 3 H]-amphetamine with rat brain synaptosomes: 
Part II. Active transport. J, Pharmacol. Exp. Ther. (in press). 

Titeler, M, Appel, N.M., De Souza, E.B. and Glennon, R.A. Receptor pharmacology of MDMA and 
related hallucinogens. Annals New York Acad. Sci. (The Neuropharmacology of Serotonin. P.M. 
Whitaker-Azmitia and S.J. Peroutka, eds.) Vol. 600:626-639, 1990. 

De Souza. E.B.. Battaglia. G. and Insel. T.R. Neurotoxic effects of MDMA on brain serotonin neurons: 
Evidence from neurochemical and radioligand binding studies. Anpals New York Acad. Sci. (The 
Neuropharmacology of Serotonin), Vol. 600:682-698, 1990. 

De Souza, E.B. Psychomimetic and neurotonic effects of amphetamines and related designer drugs. In: 
Practical Clinical Management: Drug Abuse Education for the Primary Care Physician. Medical and 
Chirurgical Faculty of Maryland , pp. 79-84, 1990. 

Kuhar. M.J. and De Souza. E.B. Receptor autoradiography as an aid in explaining drug action. In: 
Imaging the Functional Neuroanatomy of Drug Action (E.D. London, ed). The Telford Press, Caldwell, 
N.J. (in press). 

Zaczek, R., Culp, S., McCann. D. and De Souza, E.B. Sequestration of ^H-amphetamine into rat brain 
synaptosomes. American Society of Neurochemistry Meeting, Abstr. #324, 1990 

Zaczek. R.. Culp, S. and De Souza. E.B. High-affinity active transport of [ 3 H]-d-amphetamine into rat 
striatal synaptosomes. Soc. for Neurosci. . 16:303, 1990. 



35 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00303-03 NBL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: CRF in Addictive, Neuropsychiatric and Neurodegenerative disorders 

Principal Investigators: Cooperating Units 

P.I. E.B. De Souza Chief. Neurobiology Laboratory 

Others: D.E. Grigoriadis Postdoctoral Fellow. ARC 

D. Price Professor, JHU 

N. Goeders Assoc. Professor. LSU 

Cooperating Unit: Neuropathology Laboratory. JHU 

Department of Pharmacology, Louisiana State 
University Medical Center 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3.0 Professional: 2.0 Other: 1.0 

Check Appropriate Boxes: 

Human Subjects Human Tissues _X_Neither 

Minors 

Interviews 

Summary of Work 

The role of CRF in neurodegenerative diseases such as Alzheimer s disease has been examined. Changes 
in CRF and its receptor have been described. The role of CRF in neuropsychiatric disorders has been 
speculated upon as well as directly examined in patients with generalized anxiety disorder. 

This project will not be continued in 1991. 



36 



PUBLICATIONS 

McLeod. D.R., Hoehn-Saric, R., Zimmerli, W.D., De Souza, E.B., and Oliver, L.K. Treatment effects of 
Alprazolam and Imipramine: Physiological versus subjective changes in patients with generalized anxiety 
disorder. Biological Psychiatry 28:849-861. 1990. 

De Souza, E.B., Bissette, G., Whitehouse, P.J., Price, D.L., Vale, W.W. and Nemeroff, C.B. Role of 
corticotropin-releasing factor (CRF) in neurodegenerative diseases. In: Corticotropin- Releasing Factor: 
Basic and Clinical Studies of a Neuropeptide . (E.B. De Souza and C.B. Nemeroff, eds.), CRC Press, 
Boca Raton, FL., pp. 335-349, 1990. 

De Souza, E.B. Role of corticotropin-releasing factor in neuropsychiatric disorders and 
neurodegenerative diseases. Annual Reports in Medicinal Chemistry . Vol. 25 (Topics in Biology):215- 
224. 1990. 



37 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00304-03 NBL 

i 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: CRF as a Stress Neurotransmitter in the Brain-Endocrine-Immune Axis 

Principal Investigators: Cooperating Units 

P.I. E.B.DeSouza Chief, Neurobiology Laboratory 

Others: D.E. Grigoriadis Postdoctoral Fellow, ARC 

E. Webster Staff Fellow, ARC 

Cooperating Unit: None 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3.5 Professional: 2.8 Other: 0.7 

Check Appropriate Boxes: 

Human Subjects Human Tissues JL Neither 

Minors 

Interviews 

Summary of Work 

A variety of studies on CRF receptors in the brain-endocrine-immune axis have been earned out. CRF 
receptors have been identified in the mouse spleen: the cells bearing these receptors appear to be 
macrophages. Also, age-related decreases in CRF receptors have been identified in the brain and anterior 
pituitary gland of the rat. These major findings have been summarized in several publications. 

This project will not be continued in 1991. 



38 



PUBLICATIONS 

Webster, E.L., Tracey, D.E.. Jutila, M.A., Wolfe, S.A. Jr. and De Souza, E.B. Corticotropin-releasing 
factor (CRF) receptors in mouse spleen: Identification of receptor bearing cells as resident macrophages. 
Endocrinology 127:440-452, 1990. 

Heroux, J.A.. Grigoriadis, D.E. and De Souza, E.B. Age-related decreases in corticotropin-releasing 
factor receptors in the brain and anterior pituitary gland of the rat. Brain Research 542:155-158, 1991. 

De Souza, E.B. and Insel, T.R. Corticotropin-releasing factor (CRF) receptors in the rat central nervous 
system: autoradiographic localization studies. In: Corticotropin-Releasing Factor: Basic a nd Clinical 
Studies of a Neuropeptide . (E.B. De Souza and C.B. Nemeroff, eds.), CRC Press, Boca Raton, FL., pp. 
69-90, 1990. 

De Souza, E.B. and Grigoriadis, D.E. Corticotropin-releasing factor (CRF) receptors in brain: 
characterization and regulation. In: Corticotropin-Releasing Factor: Basic and Clinical Studies of a 
Neuropeptide . (E.B. De Souza and C.B. Nemeroff, eds.), CRC Press, Boca Raton, FL., pp. 115-135, 
1990. 

Battaglia, G, Webster, E.L. and De Souza, E.B. Characterization of second messengers coupled to 
corticotropin-releasing factor (CRF) receptors in brain. In: Corticotropin-Releasing Factor: Basic and 
Clinical Studies of a Neuropeptide . (E.B. De Souza and C.B. Nemeroff, eds.), CRC Press, Boca Raton, 
FL.,pp. 137-152, 1990. 

De Souza, E.B. and Appel, N.M. Distribution of brain and pituitary receptors involved in mediating 
stress responses. In: Neurobiology and Neuroendocrinologv of Stress (M.R. Brown, C Rivier and G.F. 
Koob, eds.). Marcel Dekker, Inc.. New York, pp. 91-117, 1990. 

Webster, EX., Grigoriadis, D.E. and De Souza, E.B. Corticotropin-releasing factor receptors in the brain- 
pituitary-immune axis. In: Stress. Neuropeptides, and Systemic Disease (J.A. McCubbin, P.G. Kaufmann 
and C.B. Nemeroff, eds.). Academic Press, Inc., San Diego, pp. 233-260, 1991. 

De Souza, E.B., Grigoriadis, D.E. and Webster, E.L. Role of brain, pituitary and spleen corticotropin- 
releasing factor (CRF) receptors in the stress response. In: The Stress of Life . Revisited (Methods and 
Achievements in Experiment Pathology; (G. Jasmin and M. Cantin, eds.), S. Karger, Switzerland (in 
press). 

Grigoriadis, D.E. and De Souza, E.B. Biochemical, pharmacological and autoradiography methods to 
study conicotropin-releasing factor (CRF) receptors. Methods in Neuroscience . Vol. 5:510-538, Peptide 
Technology (P.M. Conn, ed.) Academic Press, Inc., Florida, 1991. 

De Souza, E.B. Corticotropin-Releasing Hormone Receptors. In: Handbook of Chemical 
Neuroanatomy: Neuropeptide Receptors in the CNS. Part III (A. Bjorklund, T. Hokfelt and M.J. Kuhar, 
eds.), Elsevier, Amsterdam (in press). 

De Souza, E.B. Role of corticotropin-releasing factor (CRF) and its receptors in coordinating the 
endocrine, autonomic and behavioral responses to stress. European Winter Conference on Brain 
Research, Les Arcs 2000, France. 

Heroux. J.A., Grigoriadis, D.E. and De Souza, E.B. Age-related decreases in corticotropin-releasing 
factor receptors in the brain and anterior pituitary gland of the rat. Soc. for Neurosci. . 16:91, 1990. 



39 



Grigoriadis. D.E., Heroux, J.A., Pearsall, D.M. and De Souza, E.B. Biochemical isolation, 
characterization and partial purification of corticotropin-releasing factor (CRF) receptors from rat brain. 
Soc.forNeurosci. . 16:147, 1990. 



40 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00305-03 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Neurotoxicity of Selected Drugs to Monoamine Neurons in Brain 

Principal Investigators: Cooperating Units 

P.I. E.B. De Souza Chief, Neurobiology Laboratory 

Others: R. Zaczek Staff Fellow, ARC 

N.M. Appel Staff Fellow, ARC 

J.C. Contrera Pharmacologist. FDA 

Cooperating Unit: Food and Drug Administration, Rockville, MD 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.0 Professional: 1.5 Other: 0.5 

Check Appropriate Boxes: 

Human Subjects Human Tissues _Neither 

Minors 

Interviews 

Summary of Work 

The effects of selected drugs on monoamine neurons in brain has been examined. Fenfluramine, in high 
doses, has been found to decrease monoamine uptake sites in rat brain suggesting a neurotoxic effect at 
these high doses. The pharmacokinetics, and regional specificity as well as the time course of these 
effects have been examined Also, DSP-4 has been shown to have different effects on norepinephrine 
uptake in the cerebral cortex versus the hypothalamus. These studies provide evidence for a 
heterogeneity of norepinephrine sites. 

This project will not be continued in 1991. 



41 



PUBLICATIONS 

Appel. N.M., Mitchell, Wm.M. Contrera, J.F. and De Souza. E.B. Effects of high-dose fenfluramine 
treatment on monoamine uptake sites in rat brain: Assessment using quantitative autoradiography. 
Synapjss 6:33-44, 1990. 

Zaczek, R.. Battaelia. G., Culp, S., Appel, N.M., Contrera, J.F. and De Souza, E.B. Effects of repeated 
fenfluramine administration on indices of monoamine function in rat brain: Pharmacokinetics, dose 
response, regional specificity and time course data. J. Pharmacol. Exp. Ther. 253:104-1 12, 1990. 

Zaczek, R., Fritschy, J-M., Culp, S., De Souza, E.B. and Grzanna, R. Differential effects of DSP-4 on 
norepinephrine uptake into synaptosomes from cerebral cortex and hypothalamus: Evidence for 
heterotgeneity of the norepinephrine uptake sites. Srain Research 522:308-314, 1990. 

Appel, N.M., Owens. M.J., Culp, S. Zaczek, R., Contrera, J.F., Bissette, G., Nemeroff. C.F. and De 
Souza E.B. Role of brain corticotropin-releasing factor in the weight-reducing effects of chronic 
fenfluramine treatment in rats. Endocrinology (.in press). 

Appel, N.M., Zaczek, R., Mitchell. Wm.M. and De Souza. E.B. Immunohistochemical and 
autoradiographic investigations of high-dose fenfluramine treatment on monoamine neurons in rat brain. 
In: Proceedings of Int. Svmp. on Serotonin- From Cell Biology to Pharmacology and Therapeutics . (P. 
Paoletti, P.M. Vanhoutte, N. Brunello and F.M. Maggi, eds.) Kluwer Academic Publishers, Boston pp. 
625-629, 1990. 

Appel, N.M., Zaczek, R., Owens, M., Culp, S., Nemeroff, C.B. and DeSouza, E.B. Relationships 
between brain serotonin (5HT) and corticotropin-releasing hormone (CRH) in the anti-obesity effects of 
felfluramine. 1990 Winter Neuropeptide Conference, Breckenridge, Colorado, 1990. 

De Souza, E.B. Role of corticotropin-releasing factor (CRF) in the development of obesity syndromes 
and in the effects of antiobesity drugs. Winter Neuropeptide Conference, Breckinridge, Colorado, 1990. 



42 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00311-02 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: -Sigma and PCP Receptors in Neuropsychiatric Disorders 

Principal Investigators: Cooperating Units 

P.I. E.B. De Souza Chief, Neurobiology Laboratory 

Others: AD. Weissman Staff Fellow, ARC 

ED. London Chief, Neuropharmacology Lab, ARC 

M. Casanova Scientist, NIMH 

J. Kleinman Deputy Chief, Clin Brain Disorders Branch, NIMH 

Cooperating Unit: Neuropharmacology Laboratory, ARC, NIDA 
Clinical Brain Disorders Branch, 
St. Elizabeth's Hospital, NIMH 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.5 Professional: 1.3 Other: 0.2 

Check Appropriate Boxes: 

Human Subjects _X_ Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

An interesting finding has been that siema binding sites but not PCP binding sites are reduced in the 
cerebral cortex in schizophrenia However, neither of these receptors are altered in brains thought to be 
derived from PCP abusers. Chronic administration of D-pentazocine does not alter brain si^ma or 
dopamine receptors. These results suggest that there is a connection between sig ma receptors and 
schizophrenia or, possibly between sigma receptors and the treatment of schizophrenia by psychoactive 
drugs. 

This project will not be continued in 1991. 



43 



PUBLICATIONS 

Weissman, A.D.. Casanova, M.F., Kleinman, J.E., London, E.D. and De Souza, E.B. Selective reduction 
in cerebral cortical si ion a . but not PCP binding sites in schizophrenia. Biological Psychiatry 29:41-54, 
1991. 

Weissman, A.D., Casanova, M.F., Kleinman, J.E.. London, E.D. and De Souza, E.B. PCP and sigma 
receptors in brain are not altered after repeated exposure to PCP in man. Neuropsvchopharmacologv 
4:95-102, 1991. 

Weissman. AD. and De Souza, E.B. Brain sigma and dopamine receptors are not modulated by chronic 
d-pentazocine administration in rats. Eur. J. Pharmacol, (in press). 

Weissman, A.D. and De Souza. E.B. Postmortem investigations of sigma and PCP receptors in 
psychosis. In: Excerpta Medica International Congress Series (5th World Congress of Biological 
Psychiatry, Florence) 1991. 

Weissman. A.D. and De Souza. E.B. Brain sigma and dopamine receptors are not modulated by chronic 
d-pentazocine administration in rats. Soc. for Neurosci. 16:1305, 1990. 

De Souza, E.B. and Weissman. A.D. Neurotoxicity of phencyclidine and related drugs: Introduction to 
the problem and human autopsy data. American College of Neuropsychopharmacology, San Juan. Puerto 
Rico, p. 82, 1990. 



44 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00307-03 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Effects of Cocaine on Monoamines and their Metabolites in Rat Brain 
Principal Investigators: Cooperating Units 

P.I. S.Y. Yeh Pharmacologist, ARC 

Others: E.B.DeSouza Chief, Neurobiology Lab, ARC 

Cooperating Unit: None 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 5 Professional: 0.5 Other: 0.5 

Check Appropriate Boxes: 

Human Subjects Human Tissues _X_ Neither 

Minors 

Interviews 

Summary of Work 

A controversy regarding cocaine effects is whether or not chronic cocaine causes a degeneration of 
monoaminergic neurons in brain. Currently, there are some papers in the literature suggesting that such a 
neurotoxic effect occurs but many investigators have failed to replicate these findings. In a detailed 
study, we were unable to obtain any evidence for a neurochemical neurotoxicity due to repeated cocaine 
administration. These results support the notion that chronic cocaine does not cause a degeneration of 
monoamine containing neurons although many other neurotoxic effects may be possible. 



45 



PUBLICATIONS 

Yeh, S.Y. and De Souza. E.B. Lack of neurochemical evidence for neurotoxic effects of repeated 
cocaine administration in rats on brain monoamine neurons. Drug and Alcohol Dependence 27:51-61, 
1991. 

Yeh. S.Y. & De Souza. E.B. Lack of neurochemical evidence fro neurotoxic effects of repeated cocaine 
administration in rats on brain monoamine neurons. 19th meeting. Neuroscience Abstract . 1098 (1989). 



46 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00308-03 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Role of Sigma Receptors in Endocrine Organs and Immune Tissue 

Principal Investigators: Cooperating Units 

P.I. E.B. De Souza Chief, Neurobiology Laboratory 

Others: S.E. Wolfe, Jr. Staff Fellow, ARC 

Cooperating Unit: None 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.8 Professional: 1.0 Other: 0.8 

Check Appropriate Boxes: 

Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Because si gma receptors are widely distributed in the body and because a number of drugs have effects at 
sigma receptors, the localization and effects of sigma receptors in the rat pineal gland were examined. 
The localization was examined by autoradiographic methods and the effects of the drugs were explored 
with electrophysiological techniques. 

This project will not be continued in 1991. 



47 



PUBLICATIONS 

Wolfe, S.A. Jr. and De Souza, E.B. Sigma receptors in the brain-endocnne-irnmune axis. In: Sigrna, 
PCP and NMDA Receptor Systems (E.B. De Souza. E.D. London and D.H. Clouet, eds.), NED A Research 
Monographs (in press t. 

Wolfe. S.A. Jr.. Aguayo, L.G. and De Souza. E.B. Sigma receptors in rat pineal gland: 
Electrophysiology and autoradiographic localization. FASEB J. 4:A329, 1990. 



48 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00309-03 NBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Interieukin-1 in the Brain-Endocrine-Immune Axis 

Principal Investigators: 

P.I. E.B. De Souza Chief, Neurobiology Laboratory 

Others: T. Takao Visiting Fellow. ARC 

E. Webster Staff Fellow, ARC 

D.E. Tracey Visiting Scientist, Upjohn Co. 

Cooperating Unit: The Upjohn Co., Kalamazoo, MI 

Lab/Branch: Laboratory of Neurobiology, Neuroscience Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.0 Professional: 1.5 Other: 0.5 

Check Appropriate Boxes: 

Human Subjects Human Tissues JL Neither 

Minors 

Interviews 

Summary of Work 

Interleukin- 1 receptors were identified in the mouse testis as well as in the mouse brain. In the brain, the 
receptors were characterized by binding techniques and localized by autoradiographic techniques. 
Similar studies were carried out in the mouse kidney. Also, an interesting interaction between CRF 
receptors and Interleukin receptors were found in ATT- 20 tumor cells where treatment with CRF resulted 
in an upregulation of JL-1 receptors. 

This project will not be continued in 1991. 



49 



PUBLICATIONS 

Takao. T., Mitchell. Wm.M., Tracey, D.E. and De Souza, E.B. Identification of interleukin-1 receptors in 
mouse testis. Endocrinology 127:251-258, 1990. 

Takao, T., Tracey, D.E., Mitchell, Wm.M. and De Souza, E.B. Interieukin- 1 receptors in mouse brain: 
characterization and neuronal localization. Endocrinology 127:3070-3078, 1990. 

Takao, T., Mitchell. Wm.M. and De Souza, E.B. Interleukin-1 receptors in mouse kidney: 
identification, localization and modulation by lipopoiysaccaride treatment treatment. Endocrinology (in 
press). 

Cunningham, E.T. Jr., Wada, E.. Carter, D.B., Tracey, D.E., Battey, J.F. and De Souza, E.B. 
Localization of interleukin-1 receptor messenger RNA in murine hippocampus. Endocrinology (in 
press). 

De Souza. E.B.. Takao. T. and Tracey, D.E. Interleukin-1 receptors in the brain-endocrine-immune axis. 
Proceedings of the 17th C.I.N.P. Congress, Clinical Pharmacology, 1990. 

Webster. E.L., Tracey, D.E. and De Souza, E.B. Corticotropin-releasing factor (CRF) treatment 
upregulates interleukin-1 (IL-1) receptors in AtT-20 pituitary tumor cells. The Endocrine Society, 72nd 
Annual Meeting, Abstr. #379, 1990. 

Takao, T., Mitchell. Wm.M., Tracey D.E. and De Souza, E.B. Identification of Interleukin-1 receptors in 
mouse testis. The Endocrine Society, 72nd Annual Meeting, Abstr. #420, 1990. 

De Souza, E.B.. Takao, T. and Tracey, D.E. Interleukin-1 receptors in mouse brain. 2nd International 
Congress of Neuroendocrinology. Neuroendocrinolog v Vol. 52(S1):76, 1990. 

Wada, E., Cunningham. E.T. Jr., Mitchell, Wm.M., Carter. D.B., Tracey, D.E., Battey, J.F. and De 
Souza, E.B. Identification of interleukin-1 receptor mRNA in murine hippocampus. Soc. for Neurosci. . 
16:1213, 1990. 

Takao, T., Tracey, D.E.. Mitchell. Wm.M. and De Souza, E.B. Interleukin-1 receptors in mouse brain: 
Characterization and autoradiographic localization. Soc. for Neurosci . 16:1213, 1990. 

De Souza, E.B., Takao. T. and Tracey, D.E. Interleukin-1 receptors in the brain-endocrine-immune axis. 
American College of Neuropsychopharmacology, San Juan, Puerto Rico. p. 66, 1990. 



50 



4. Molecular Neurobiology Laboratory - George R. Uhl, M.D., Ph.D., Chief 

Overview 

Drugs impact the nervous system through interactions with the products of specific genes, some 
characterized and others unknown. The Laboratory of Molecular Neurobiology, formed within the 
Neuroscience Branch in 1989, studies the structure and regulation of genes involved in the actions of 
abused drugs. The laboratory studies how drugs and neural function change gene regulation in the brain, 
the structures of the genes encoding drug and neurotransmitter receptors, and genetic differences in drug- 
abusing populations. Insights derived from these studies are applied in human clinical studies. 

A principal working hypothesis motivating this work is that the mechanisms regulating genes of 
neurotransmission can reflect, or store, information about prior brain exposure to drugs or other stimuli. 
The laboratory thus studies specific patterns of brain regulation of genes for neurotransmitters impacted 
by abused drugs within specific neural populations in the brain. Genes for neurotransmitters that are 
modulated by abused substances such as preproenkephalin. genes for transcription factors that could be 
involved in this modulation, and genes expressed in key neurons of central brain pathways for 
reinforcement reward are studied in normal and transgenic animals. Knowledge of these regulated 
mechanisms guides the formulation of strategies for directly influencing gene expression in neurons, and 
clinical studies identifying whether such mechanisms can be discerned and manipulated in man. 

A second major hypothesis is that better understanding of the detailed molecular structures of the 
receptor molecules for abused drugs, improved ability to develop anti-abuse medications working at these 
receptors, and ability to study gene abnormalities in drug-abusing populations will come through cloning 
receptor genes. The laboratory uses homology and expression strategies to elucidate the structures of 
receptor cDNAs and genes. 

Summary of Ongoing Research 

A. Receptor and Receptor-Modulating cDNAs 

Identifying genes encoding the cell surface receptors for abused drugs is an important step in the 
molecular biology of drug abuse. Since these rare membrane proteins are difficult to purify through 
conventional means, the laboratory has utilized and developed DNA homology and expression cloning 
approaches for gene identification and characterization 

Ligand-Gated Channels: GABA Rho and Others 

Polymerase-chain reaction homology approaches utilize the similarity between receptor families to 
identify cDNAs with receptor-like properties. A novel benzodiazepine/ barbiturate/GABA receptor 
subunit cloned in collaborative studies including this laboratory displays interesting properties on 
expression that correspond to those previously described of the "GABA C" receptor. The robust 
electrophysiological responses of this receptor, expressed as a single subunit, make it an ideal backbone 
on which to superimpose the pharmacological and physiological properties conferred by domains of other 
GABA A receptor subunits. These approaches could allow more precise molecular definition of the 
poorly-understood sites for barbiturate and benzodiazepine drug actions. 

PCR approaches have also defined other novel receptor subtypes that may he in the inhibitory amino acid 
receptor famdy, and in the excitatory amino acid receptor family that includes the phencyclidine 
receptor. 



51 



G- Linked Receptor Expression and Modulation: Xenopus Oocytes and COS Cells 

Cholecystokin (CCK) is a dopamine cotransmitter intimately involved with VTA circuits key to central 
mechanisms of reward and reinforcement. Subfractionation of libraries by sib selection has allowed 
isolation of a cDNA whose expression can lead to CCK responses when injected into oocytes by 
collaborators. The nature of these responses is consistent with a CCK-A receptor. The clone that confers 
these pharmacologically- and physiologically-appropriate responses has been sequenced, and its 
expression in brain noted in Northern analyses. The cDNA predicts an open reading frame that has a 
unique sequence, is not of the seven transmembrane class, but may be a modifier of the expression of an 
endogenous Xenopus CCK receptor gene regulator by antisense mechanisms. 

The laboratory has defined strategies for expression of the cocaine receptor and the endothelin receptor in 
the Xenopus system, and has utilized COS cell transient expression in conjunction with ligand binding to 
allow exploration of the function of other unknown receptor-like cDNA clones. 

Approaches to Transporter Cloning: Cocaine Receptor/Dopamine Transporter cDNA Sought by 
Oocyte Expression, PCR, LARS 

The laboratory has taken three distinct approaches to cloning the cocaine receptor/dopamine transporter. 
Expression in Xenopus oocytes allows detection of uptake of radiolabeled dopamine in oocytes injected 
with mRNA prepared from tissue rich in dopaminergic cell bodies. The pharmacological and 
physiological specificity of transport is good, but the assay s sensitivity is variable. In a second approach, 
a newly-developed technique for ligand autoradiographic receptor screening is utilized. This technique 
was validated using beta adrenergic receptor expression, which documented that cDNA enrichments of 
300-fold could be achieved in a single round of screening. These workers have tested a number of 
cocaine receptor radioligands for activity in this system, and have used an iodinated cocaine analog to 
screen libraries for expression of this receptor. In a third approach, the laboratory has used the 
transmembrane sequences that appear to be conserved between the GABA and norepinephrine 
transporters to clone PCR products and cDNAs belonging to the 1 2-transmembrane domain famdy of 
sodium-dependent transporters. 

B. Neuronal Expression of Genes for Neurotransmitters Related to Drug Abuse 

Understanding the ways in which neurons regulate their expression of the neurotransmitter genes that are 
related to drug abuse can help to elucidate neurochemical mechanisms for drug action, including 
mechanisms of tolerance and dependence. The laboratory has defined changes in the neuronal expression 
of several genes potentially related to drug action, and identified specific transcription factor pathways 
well positioned to play key roles in proenkephalin regulation in vivo . Transgenic animals that have been 
developed and characterized in the laboratory support a role for specific regulatory elements in tissue- 
specific expression, and in some but not in other examples of trans-synaptic gene regulation. Modified 
oligonucleotides that could help to experimentally regulate gene expression have been synthesized, and 
display remarkable abilities to enter and persist in brain neurons in largely-undegraded form that could 
act to regulate gene expression in a directed fashion. 

Neuronal Expression of Neurotransmitter Genes in Central Brain Pathways of 
Reinforcement/Reward 

Specific neuronal pathways in the brain are selectively and importantly implicated in the reinforcing and 
rewarding properties of a number of abused drugs, especially cocaine. The laboratory has examined key 
ventral tegmental area neurons of this circuitry, and found a substantial diversity of gene expression 
among the different subnuclei of neurons implicated in these processes. These studies complement 
ongoing clinical studies of drug effects in patients with lesions in VTA neurons. 



52 



PUBLICATIONS 

Jayaraman, A., Nishimori. T., Dobnar, P. and Uhl, G. Cholecystokinin and neurotensin mRNAs are 
differentially expressed in subnuclei of the ventral tegmental area. J. Comp. Neuro. 296: 291-302, 1990. 

Rattray, M., Lautar, S.L. and Uhl, G.R. Ligand autoradiographic receptor screening: receptor cDNA 
expression in replicas of transfected COS cells. Mol. Br. Res, 7(3): 249-259, 1990. 

Uhl, G.R. Parkinsons disease: neurotransmitter and neurotoxin receptors and their genes. Eur. J. 
Nejupl 30(1): 21-30, 1990. 

Uhl G. Neurotensin receptors. In: A. Bjorklund, T. Hokfelt, M. Kuhar (Eds.): Handbook of Chemical 
Neuroanatomy. Vol. IX, . Elsevier, New York, 1990, pp. 443-453. 

Price, D.L., Whitehouse, P.J., Struble. R.G., Hedreen, J.C. and Uhl. G.R. Transmitter systems in 
selected types of dementia. In: P. Riederer, N. Kopp and J. Pearson (Eds): An Introduction to 
Neurotransmission in Health and Disease . Oxford University Press. New York, 1990, pp. 349-357. 

Uhl, G.R. Principles of assessing neurotransmitter receptors in disease. In: J. James Frost and Henry N. 
Wagner. Jr.. (Eds.): Quantitative Imap i np- Neuroreceptors. Neuroreceptors. Neurotransmitters, and 
Enzymes. Raven Press. Ltd., New York. 1990, pp. 9-18. 

Uhl. G.R.. Messenger mRNA localization with the microscope: in situ hybridization using radiolabeled 
probes. In: Yamamura, Enna and Kuhar (Eds.): Methods in Neurotrans mitter Receptor Analysis . Raven 
Press, New York, 1990, pp. 219-244. 

Uhl, G.R. and Nishimori, T. Neuropeptide gene expression and neural activity: assessing a working 
hypothesis in nucleus caudalis and dorsal hom neurons expressing preproenkephalin and 
preprodynorphin. Cell, and Mol. Neurobiology. 10(6): 73-98, 1990. 

Nishimori, T., Buzzi, M.G., Chudler, E.H., Poletti, C.E., Moskowitz, M.A. and Uhl, G.R. 
Preproenkephalin upregulation in nucleus caudalis: high and low intensity afferent stimulation 
differentially modulate early and late responses. J. Comp. Neurol. 302: 1002-1018, 1990. 

Articles in Press 

Uhl, G.R., O'Hara, B.. Shimada. S., Zacek, R., DiGiorgianni. J. and Nishimori, T. Dopamine transporter: 
expression in Xenopus oocytes. Mol. Br. Res. . In press. 

Uhl. G.R., Jayaraman. A., Nishimori, T., Shimada, S., Rattray, M. and O'Hara, B. Neuropharmacologic 
techniques in the molecular biology of schizophrenia. In: SC Schultz and CA Tamminga (Eds.): 
Schizophrenia Research. Advances in Neuropsychiatry and Psvchophan nacologv. Vol. 1. Raven Press, 
Ltd., New York, In press. 

Uhl, G.R., Appleby, DA. and Nishimori T.N. Regulated expression of transcription factor and 
neurotransmitter genes in neural populations: studies using in situ hybridization with radiolabeled 
oligonucleotides probes. In: A. Calas and D. Eugene (Eds.): Recent Advances in NeurpcytQchemKal 
Methods . Springer- Verlag, Berlin-Heidelberg, In Press. 

Schaeffer, J.S., Lin, C-L, Kitayama S. and Uhl, G.R. Ligand autoradiographic receptor screening II: 
expression of receptor cDNA in transfected COS cells grown on polyester disks and its recovery. Mol. 
Rr^in Res . In press. 



53 



Shimada, S., Spivak, C. and Uhl, G. Endothelin receptor: A profoundly desensitizing receptor expressed 
in Xenopus oocytes. Eur. J. of Pharmacology . In press. 

Cutting, G.R.. Lu. L., O'Hara. B.F.. Kasch, L.M., Montrose-Rafizadeh, C, Donovan. D.M., Shimada. S., 
Antonarakis. S.E.. Guggino, W.B., Uhl, G.R. and Kazazian, H.H. Cloning of the q-aminobutyric 
acid (GABA) p[ cDNA: a GABA receptor subunit highly expressed in the retina. Proc. Natl. Acad. Sci. . 
In press. 

O'Hara, B.F., Smith, S.S., Persico, A., Wang, K., Cutting, G.R.. Newlin, D.B., Gorehck. DA. and Uhl, 
G.R. Dopamine D2 receptor alleles in substance abusers: confounding effect of race. JAMA . In press. 

Uhl. G.R., Walther. D., Nishimori, T.. Buzzi. G. and Moskowitz, M. Jun B, CJun. Jun D and cFOS 
mRNAs in nucleus caudalis neurons: Rapid selective enhancement by afferent stimulation. Mol. Brain 
Res., In press. 

Stopa, E.G., Uhl, G.R., Mobtaker, H., Winnepenny, R., Hoefler, H., King, J.C.. Bird, E.D. and Wolfe. H. 
Somatostatin-gene expression in human brains: in situ hybridization studies in postmortem tissue. V, 
Arch. Path.. In press 

Uhl, G.R., Appleby, D., Nishimori. T.. Buzzi, M.G. and Moskowitz, M.A. Synaptic regulation of the 
enkephalin gene and transcription factors in vivo: possible roles in drug abuse. NIDA Research 
Monograph. Problems of Drug Dependence. 1990. Proceedings of the 52nd A nni l al Scientific Meeting- 
Committee on Problems of Drug Dependence. Inc. . In press. 

Uhl, G.R. Identifying and localizing gene expression important for the actions of abused drugs. In: 
Imping the Functional Neuroanatomy of Drug Action. Telford Press, In press. 



Abstracts 

Jayaraman, A. and Uhl. G.R. Cholecystokinin (CCK) mRNA: Prominent expression in the rostral nigral 
neurons. Movement Disorders 5( 1): 48. 1990. 

Uhl, G.R., Shimada, S., O'Hara, B.. DiGiorgianni. J., and Nishimori. T. Dopamine transporter mRNA 
and cDNA: Strategy for expression cloning a selective neurotoxin concentrator. Movement Disorders 
5(1): 89, 1990. 

Nishimori, T., Buzzi, G, Moskowitz, M.A., Ross, C, Snyder, S.H. and Uhl, G.R. Enkephalin convertase 
gene expression in rat nucleus caudalis neurons. Neurobiology of Nociception Abstract. 1990. 

Uhl, G.R.. Newlin, D.B., Pretorius. M.B., Park. J.S.. Darwin, W.D. and Cone, E. Antagonist-withdrawal 
up-regulation of endogenous opiate antinociceptive systems. CPpn Annual .Scientific Abstract. 1990. 

Uhl, G.R.. Buzzi. M.G., Appleby, D., Moskowitz, M.A. and Nishimori, T. Transcription factors binding 
to the preproenkephalin promoter: expression in neurons of normal and stimulated nucleus caudalis. 
Society for Neuroscience Abstracts 16(2): 1275, 1990. 

Donovan, D.M., Cutting, G.R., O'Hara, B.F., Shimada, S.S. and Uhl, G.R. Receptor families: cDNAs 
identified by PCR & DNA homologies. Society for Neuroscience Abstracts 16(2) 1018, 1990. 

O'Hara, B.F., DiGiorgianni, J.M., Shimada, S., Landau, E.M.. Uhl, G.R. and Meiri, H. NT and CCK 
receptor cDNAs: enrichment by oocyte expression and cDNA library sib selection. Society for 
Neuroscience Abstracts 16(1): 82, 1990. 



54 



Shimada, S. and Uhl. G.R. Endothelin receptors: expression in Xenopus oocytes. Society for 
Neuroscience Abstracts 16(1): 686, 1990. 

Uhl, G.R., Appleby, D. and Nisbimori, T. Can we deduce mechanisms for neurotransmitter gene 
regulation in situ ? The enkephalin promoter "cassette" and trans-synaptic transcription factor u- 
regulation. Third Research Symposium of the Center for Studies in Reproduction. Molecular and 
Cellular Advances in Endocrinology . Univ. Of Maryland School of Medicine, Oct. 1-2, 1990. 

Uhl, G.R. Parkinsons disease: neurotransmitter and neurotoxin receptors and their genes. 
Parkinson/ Alzheimer Digest 5: 20-21, 1990. 



Abstracts in Press 

Uhl, G.R.. O'Hara, B.F.. Smith. S.S.. Persico, A., Newlin, D.B. and Gorelick. D2 receptor alleles in drug 
abusers: confounding effects of race. CPDD Annual Scientific Meeting, in press. 

Uhl, G.R., Appleby, D., Buzzi, M.G. and Moskowitz, M. Dorsal horn proenkephalin regulation by 
primary afferents: co-regulation of specific transcription factor genes. Neurology. In press. 

Jayaraman. A. and Uhl, G. Specific subpopulation of substantia nigra pars compacta neurons express the 
CCK gene. Neurology . In press. 



55 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 001 14-02 MNL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: 

Receptor cDNA Expression Cloning Using Ligand Autoradiographic Screening 

Principal Investigators: Cooperating Units 

Uhl, G.R.. Chief Laboratory of Molecular Neurobiology, ARC. 

Others: Rattray, M.A.N. , Foreign Fellow. MPL, ARC; Lautar, S., Research Lab Manager, MPL. ARC: 
Lin, Glen, Research Technician (guest worker) Dept. of Neuroscience, Johns Hopkins University School 
of Medicine, Baltimore. MD 21205. 

Lab/Branch: 

Laboratory of Molecular Neurobiology, Neuroscience Branch 

Section: 

Gene Neuroscience Unit. 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2 Professional: 1 Other: 1 

Check Appropriate Boxes: 

Human Subjects _ Human Tissues _X Neither 

Minors 

Interviews 

Summary of Work: 

Understanding the receptor molecules that recognize abused drugs and the neurotransmitters impacted by 
drugs is an important step in determining the molecular mechanisms underlying drug abuse. Little is 
known about many of these molecules, because they have proven very difficult to purify through 
conventional approaches. The laboratory has continued to work to establish a method for direcdy cloning 
these molecules based on their ligand binding properties, and to exploit this approach. 

Over the past year, these workers have made substantial progress toward this end. Previously, a cloned 
beta adrenergic receptor cDNA was used to document and optimize expression of the beta adrenergic 
receptor binding site in COS cells. In studies ompleted during this year, improved DNA recovery 
techniques and electroporation have resulted in recoveries of the plasmid present in very small number of 
cells with a reasonable frequency. This results in documented enrichments of up to 300-fold in a single 
step. The workers have progressed in adapting the method for use with the cocaine and PCP receptors. 
Goals for the current year include using this technique to attempt receptor cloning based on both cDNA 
recovery and subfractionating libraries and adaptation of the polymerase chain reaction to aid in recovery 
of the very small numbers of plasmids present. This approach continues to provide promise for allowing 
direct cloning of genes key to the action of several classes of abused substances. 



56 



PUBLICATIONS 

Rattray. M.. Lautar. S.L. and Uhl. G.R. Ligand autoradiographic receptor screening: receptor cDNA 
expression in replicas of transfected COS cells. Mol. Br. Res. 7(3): 249-259. 1990. 

Schaeffer, J.S., Lin. C-L. Kitayama. S. and Uhl, G.R. Ligand autoradiographic receptor screening II: 
Expression of receptor cDNA in transfected COS cells grown on polyester disks and its recovery. Mol. 
Brain Res. . In press. 



57 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00115-03 MNL 

Period Covered: January 1, 1990 to December 31, 1990 
Title of Project: 

Receptor cDNA Expression Cloning Using Xenopus Oocyte Expression 

Principal Investigators: Cooperating Units 

Uhl. G.R.. Chief Laboratory of Molecular Neurobiology, ARC 

Others: Shimada. S., Visiting Scientist, LMN, ARC , O'Hara, B., Staff Fellow, LMN, ARC and 
DiGiorgianni, J., Technician (guest worker), Johns Hopkins University, Dept. of Neuroscience, Johns 
Hopkins School of Medicine; Spivak C Pharmacologist , NPL, ARC, Drs. Suzanne Zukin and Michael 
Bennett, Albert Einstein College of Medicine, Dr. Emmanuel Landau, Mt. Sinai School of Medicine, 
New York. 

Neuropharmacology Laboratory, ARC. 

Lab/Branch: 

Laboratory of Molecular Neurobiology, Neuroscience Branch 

Section: 

Gene Neuroscience Unit. 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 15 Professional: 1 Other: 0.5 

Check Appropriate Boxes: 

Human Subjects Human Tissues _X Neither 

Minors 

Interviews 

Summary of Work: 

Expression cloning is an attractive approach to identifying the genes and cDNAs encoding receptors for 
abused substances and for the neurotransmitters implicated in drug abuse (see above). The ability of the 
Xenopus oocyte to appropriately translate, post-translationally modify, and appropriately insert several 
receptors into its membrane has led us to establish this system as a screening tool for these cDNAs and 
receptors. 

Progress in this project has included: 1) Obtaining electrophysiologic signals for neurotensin, 
cholecystokinin. and kainic acid from oocytes injected with these synthetic mRNA transcripts, 2) 
Cloning of a single cDNA that induces CCK-A responses in oocytes that are pharmacologically- 
appropriate, but likely due to upregulation of the endogenous CCK receptor gene by the injected 
transcript. This cDNA "CCK-UP" is expressed in several brain regions, and may provide important 
insights into the upregulation mechanisms important for one of the chief dopamine cotransmitters in the 
brain. 3) Characterizing interesting properties of the GABA-Rhol receptor expressed in this system, 
including bicucculing-insensitivity. This approach thus enhances abilities to clone intercstingc DNAs 



58 



through "sib selection" techniques, and improves abilities to characterize the properties of expressed 
cDNAs. 

PUBLICATIONS 

Uhl, G.R., O'Hara. B., Shimada, S., Zacek. R., DiGiorgianni, J. and Nishimori, T. Dopamine 
Transporter: Expression in Xenopus oocytes. Mol. Br. Res. . In press. 

Shimada. S., Spivak. C. and Uhl. G. Endothelin receptor: A proundly desensitizing receptor expressed 
in Xenopus oocytes. Eur. J. of Pharmacology. In press. 

Cutting, G.R., Lu, L., O'Hara. B.F., Kasch, L.M., Montrose-Rafizadeh, C, Donovan, DM., Shimada. S., 
Antonarakis, S.E., Guggino, W.B., Uhl, G.R. and Kazazian, H.H. Cloning of the q-aminobutyric acid 
(GABA) p-aminobutyric acid (GABA) p i cDNA: a GABA receptor subunit highly expressed in the retina. 
Proc. Natl. Acad. Sci. . In press. 

Uhl, G.R.. Shimada, S., O'Hara. B., DiGiorgianni, J., and Nishimori, T. Dopamine transporter mRNA 
and cDNA: Strategy for expression cloning a selective neurotoxin concentrator. Movement Disorders 
5(1): 89, 1990. 

Donovan, D.M., Cutting, G.R.. O'Hara, B.F., Shimada, S.S. and Uhl, G.R. Receptor Families: cDNAs 
identified by PCR & DNA homologies. Society for Neuroscience Abstracts 16(2): 1018, 1990. 

O'Hara, B.F.. DiGiorgianni, J.M., Shimada, S., Landau. E.M., Uhl, G.R. and Meiri, H. NT and CCK 
receptor cDNAs: Enrichment by oocyte expression and cDNA library sib selection. Society for 
Neuroscience Abstracts 16(1): 82, 1990. 

Shimada. S. and Uhl, G.R. Endothelin receptors: Expression in Xenopus oocytes. Society for 
Neuroscience Abstracts 16(1): 686. 1990. 



59 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00116-03 MNL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: 

Genes Related to Drug Abuse I: Regulation of Opioid Peptide Genes. 

Principal Investigators: Cooperating Units 

Uhl, G.R. Others: Appleby, D., LMN. Research Technician. ARC/NIDA, DiGiorgianni, J., Technician 
(guest worker); Moskowitz, M., Associate Professor of Neurology & Neurosurgery*. 

* Departments of Neurology, Neurosurgery, Massachusetts General Hospital, Harvard Medical School 
and Department ofNeuroscience. Johns Hopkins University School of Medicine. 

Lab/Branch: 

Molecular Neurobiology Laboratory, Neuroscience Branch 

Section: 

Gene Neuroscience Unit 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.0 Professional: 1 Other: 1 

Check Appropriate Boxes: 

_X Human Subjects Human Tissues _ Neither 

Minors 

Interviews 

Summary of Work: 

Drugs alter the expression of many important genes in the brain. These changes in gene expression are 
likely to contribute to long-term drug effects, such as tolerance and dependence. Continuing studies of 
the genes encoding endogenous morphine-like peptides suggest that drug-induced gene regulation 
depends on the circuits connecting to the neuron and the duration of drug exposure. 

During the past year, the laboratory has monitored cellular levels of neuropeptide mRNAs and of the 
transcription factor genes that may alter expression of these mRNAs to assess gene regulation related to 
functional activity in these neurons. In studies completed during this year, transsynaptic regulation of 
preproenkephalin expression in pain-modulating neurons of the nucleus caudalis of the spinal tract of the 
trigeminal was described in detail, and the AP-1 and CREB family transcription factors that could 
participate in this regulation were identified. Transgenic mice possessing a specific portion of the 
preproenkephalin promoter were produced, and their responsiveness to only certain trans-synaptic 
activators of preproenkephalin expression noted. Interesting effects of expressed minigene constructs on 
male fertility and testicular function were found. Finally, approaches to directly modifying these 
transcription factor pathways in brain with intraparenchymal injection of modified oligonucleotides were 
validated. These approaches allow understanding of mechanisms important for attempts to 
therapeutically manipulate drug influences on gene expression. Preliminary results from a human study 
using opiate antagonists to change the expression of these opioid peptide genes provide evidence for the 



60 



efficacy of such targeted therapies. 



PUBLICATIONS 

Uhl, G.R. Messenger mRNA localization with the microscope: in situ hybridization using radiolabeled 
probes. In: Yamamura, Enna and Kuhar (Eds.): Methods in Neurotransmitter Receptor Analysis . Raven 
Press, New York, 1990, pp. 219-244. 

"Uhl, G.R. and Nishimori, T. Neuropeptide gene expression and neural activity: Assessing a working 
hypothesis in nucleus caudalis and dorsal horn neurons expressing preproenkephalin and 
preprodynorphin. Cellular and Molecular Neurobiology 10(6): 73-98, 1990. 

Nishimori. T., Buzzu. M.G., Chudler, E.H., Poletti. C.E., Moskowitz. M.A. and Uhl. G.R. 
Preproenkephalin upregulation in nucleus caudalis: high and low intensity afferent stimulation 
differentially modulate early and late responses. J. Comp. Neurol . 302: 1002-1018, 1990. 

Uhl, G.R., Jayaraman. A., Nishimori. T., Shimada. S.. Rattray, M. and O'Hara. B. Neuropharmacologic 
techniques in the molecular biology of schizophrenia. In: S.C. Schultz and C.A. Tamminga (Eds.): 
Schizophrenia Research. Advances in Neuropsychiatry and Psvchopharmacologv. Raven Press, Ltd.. 
New York, In press. 

Uhl, G.R., Appleby, DA. and Nishimori. T.N. Regulated expression of transcription factor and 
neurotransmitter genes in neural populations: Studies using in situ hybridization with radiolabeled 
oligonucleotides probes. In: A. Calas and D. Eugene (Eds.), Recent Advances in Neurocvtochemical 
Methods . Springer- Verlag, Berlin-Heidelberg, In press. 

Uhl. G.R., Walther, D., Nishimori. T., Buzzi, G. and Moskowitz, M. Jun B. cJun. Jun D and cFOS 
mRNAs in nucleus caudalis neurons: Rapid selective enhancement by afferent stimulation. Mol. Brain 
Res. . In press. 

Uhl, G.R., Appleby, D., Nishimori, T., Buzzi. M.G. and Moskowitz, M.A. Snynaptic regulation of the 
enkephalin gene and transcription factors in vivo: possible roles in drug abuse. NIDA Research 
Monograph. Problems of Drug Dependence . 1990, Proceedings of the 52nd Annual Scientific Meeting, 
Committee on Problems of Drug Dependence, In press. 

Jayaraman, A. and Uhl. G.R. Cholecystokinin (CCK) mRNA: Prominent expression in the rostral nigral 
neurons. Movement Disorders 5(1): 48, 1990. 

Nishimori. T., Buzzi, G., Moskowitz. M.A., Ross, C, Snyder, S.H. and Uhl, G.R. Enkephalin convertase 
gene expression in rat nucleus caudalis neurons. Neurobiology of Nociception Abstract . 1990. 

Uhl, G.R.. Newlin, D.B., Pretorius, MB.. Park, J.S., Darwin, WD. and Cone, E. Antagonist-withdrawal 
up-regulation of endogenous opiate antinociceptive systems. CPDD An nual Scientific Abstract. 1990. 

Uhl, G.R., Buzzi. M.G.. Appleby, D., Moskowitz, M.A. and Nishimori, T. Transcription factors binding 
to the preproenkephalin promoter: expression in neurons of normal and stimulated nucleus caudalis. 
Society for Neuroscience Abstracts 16(2): 1275, 1990. 

Uhl, G.R., Appleby, D. and Nishimori, T. Can we deduce mechanisms for neurotransmitter gene 
regulation in situ ? The enkephalin promoter "casette" and trans-synaptic transcription factor up- 
regulation. Third Research Symposium of the Center for Studies in Reproduction . Molecular and 
Cellular Advances in Endocrinology . Univ. of Maryland School of Medicine, Oct. 1-2, 1990. 



61 



Utal. G.R. Parkinsons disease: neurotransmitter and neurotoxin receptors and their genes. 
Parki nson/Alzheimer Digest 5: 20-21. 1990. 

Uhl. G.R., Appleby, D., Buzzi. M.G. and Moskowitz, M. Dorsal hom proenkephalin regulation by 
primary afferents: co-regulation of specific transcription factor genes. Neurology . In press. 

Jayaraman. A. and Uhl, G. Specific subpopulation of substantia nigra pars compacta neurons express the 
CCK gene. Neurology , In press. 



62 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00117-02 MNL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: 

Genes Related to Drug Abuse II: Central Brain Pathways of Reinforcement/Reward. 

Principal Investigators: Cooperating Units 

Uhl. G.R. Others: Appleby, D., Research Technician. LMN. ARC NTDA. Rao, J., Dept. of Neurology, 
Louisiana State University, New Orleans, LA. 

Lab/Branch: 

Molecular Neurobiology Laboratory, Neuroscience Branch 

Section: 

Gene Neuroscience Unit 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2 Professional: 1 Other: 1 

Check Appropriate Boxes: 

_X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work: 

Neurons in the ventral tegmental area are major constituents of central brain pathways of drug 
reinforcement. Despite this potential central role in the actions of many abused drugs, however, the 
detailed expression of important neurotransmission genes in these neurons has not been elucidated The 
expression of the genes encoding two of the dopamine-cotransmitter peptides, cholecystokinin and 
neurotensin, have thus been mapped to neurons in this area during this year. In these studies, a surprising 
region-to-region variation in the expression of neurotensin and CCK genes in these neuronal subdivisions 
has been found. Such detailed studies are necessary before defining activities of cocaine and other 
abused drugs on this expression. These approaches thus allow detailed examination of the ways in which 
drugs and other physiologic processes influence gene regulatory mechanisms that could serve as a store 
for some of the information about prior drug use that may accompany tolerance and dependence. 

The potential function of these neurons in human drug-induced reinforcement/reward is being studied as 
well. Parkinson's disease patients, whose brains lose VTA neurons, are assessed after administration of 
methylphenidate. Blunting of drug effects on mood in these patients would support a central role for 
these neurons in reinforcement/reward in man. 



63 



PUBLICATIONS 

Uhl G.R. Parkinsons disease: Neurotransmitter and neurotoxin receptors and their genes. European J. 
Neurol . 30(1): 21-30, 1990. 

Jayaraman, A., Nishimori, T., Dobnar, P. and Uhl, G. Cholecystokinin and Neurotensin mRNAs are 
differentially expressed in subnuciei of the ventral tegmental area. J. Comp. Neurol. 296: 291-302, 1990. 

Uhl, G. Neurotensin receptors. In: A. Bjorklund. T. Hokfelt. M. Kuhar (Eds.). Handbook of Chemical 
Neuroanatomy, Vol. IX. Elsevier. New York, 1990, pp. 443-453. 

Price, D.L., Whitehouse, P.J., Struble, R.G., Hedreen. J.C. and Uhl. G.R. Transmitter systems in selected 
types of dementia. In: P. Riederer, N. Kopp and J. Pearson (Eds). An Introduction to Neurotransm ission 
in Health and Disease. Oxford University Press, New York, 1990, pp. 349-357. 

Uhl. G.R. Principles of assessing neurotransmitter receptors in disease. In: J. James Frost and Henry N. 
Wagner (Eds). Ouanmativ e Imafnnp ; Neuroreceptors. Ne urotrans mitters, and Enzymes . Raven Press. 
Ltd., New York. 1990. pp. 9-18. 

Stopa, E.G.. Uhl. G.R.. Mobtaker. H.. Winnepenny. R.. Hoefler. H.. King, J.C, Bird. E.D. and Wolfe, H. 
Somatostatin-gene expression in human brains: in situ hybridization studies in postmortem tissue. V. 
Arch. Path.. In press. 



64 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00314-01 MNL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: 

D2 Receptor Gene Allelic Association with Substance Use 

Principal Investigators: Cooperating Units 

Uhl, G.R., Chief Laboratory of Molecular Neurobiology, ARC 

Others: O'Hara, B., Staff Fellow, Laboratory of Molecular Neurobiology, ARC; Fanner, S., Laboratory 
Technician, Laboratory of Molecular Neurobiology, ARC; Persico, A., Visiting Fellow, Laboratory of 
Molecular Neurobiology, ARC; Corporating Units: Smith. S., Staff Fellow, Etiology Branch. ARC; 
Newlin. D., Acting Branch Chief, Etiology Branch, ARC; Gorelick, D., Chief, Treatment Branch, ARC. 

Lab/Branch: 

Laboratory of Molecular Neurobiology, Neuroscience Branch 

Section: 

Gene Neuroscience Branch 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.5 Professional: 1.5 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

X Interviews 

Summary of Work: 

Recent observations that the dopamine D2 receptor gene Taq I RFLP may be associated with alcoholism 
have motivated attempts to test whether or not this geneotype is associated with an increased incidence of 
drug abuse. Over the past year, these workers have made substantial progress toward an initial test of the 
possible allelic association between the Al allele and self-reported drug use. DNAs from approximately 
160 individuals from the Addiction Research Center, Francis Scott Key Medical Center, and Johns 
Hopkins School of Medicine Genetics clinic have been analyzed. A cDNA subclone that improves the 
assay has been constructed. A racial association with the Al allele has been identified, and self-reported 
use of a number of different drugs have been assessed for possible linkage with the Al allele. Goals for 
the current year include: Assaying whether individuals with different genotypes have different responses 
to methylphenidate, following up on any drug-alleleo associations, and considering the role of the 
Laboratory of Molecular Neurobiology in larger-scale genetic linkage studies. 



65 



PUBLICATIONS 

O'Hara. B.. Smith. S., Persico, A., Wang, K.. Cutting, G., Newlin, D., Gorelick, D. and Uhl. G.R. 
Dopamine D2 receptor alleles in substance abuses: confounding effect of race. JAMA (Submitted). 



66 



Preclinical Pharmacology Branch 

Steven R. Goldberg, Ph.D.. Chief 

Introduction 

The Preclinical Pharmacology Branch conducts research in experimental animals on the behavioral 
modes of action of drugs of abuse both in producing reinforcing, punishing and discriminative stimuli and 
in altering established behavior controlled by non-drug events such as food or electric shock. It also 
studies the role of genetics in determining the effects of drugs of abuse. The overall goal of the Branch is 
to understand how drugs of abuse affect whole animals, with an eventual goal being the discovery of 
effective treatment regimens. Studies cover a wide range of topics, including the pharmacology of 
opioids, psychomotor stimulants, and benzodiazepine dependence, alterations in the acquisition and 
retention of classically conditioned behavioral and physiological responses by drugs of abuse, 
consequences of repeated drug administration, and environmental and genetic determinants of drug- 
seeking and drug-taking behavior. Drugs are also evaluated to characterize physiological and toxic 
actions which accompany acute and prolonged administration, delineate the mechanisms responsible for 
these effects and establish methods for preventing or reversing the effects. 

Research is carried out in both nonhuman primates (rhesus and squirrel monkeys) and non-pnmates. 
New drugs are evaluated for abuse potential by comparison of reinforcing, aversive and discriminative 
stimulus effects, and by comparison of effects of prototypic drugs of abuse on neurophysiologic systems. 
Emphasis is placed on the use of pharmacological, environmental and genetic interventions to alter the 
effects of drugs of abuse. These aims are intended to further the overall goals of the Addiction Research 
Center by providing a background of information to be used in developing rational clinical procedures for 
the prevention and treatment of drug abuse. 

The Preclinical Pharmacology Branch consists of two laboratories, a Behavioral Pharmacology and 
Genetics Laboratory and a Psychobiology Laboratory. The two laboratories are subdivided into 
functional and collaborative units with emphasis on behavioral pharmacology, drug self-administration, 
physiological psychology, pharmacogenetics, behavioral and biochemical genetics, medications 
development and neuropsychopharmacology and toxicity. 

1. Behavioral Pharmacology and Genetics Laboratory - Steven R. Goldberg, Ph.D., Chief 

Overview 

The Behavioral Pharmacology and Genetics Laboratory is responsible for research in experimental 
animals on environmental, historical and pharmacological determinants of the reinforcing and other 
behavioral effects of drugs of abuse and on their physiological effects including toxicity. Drugs of abuse 
from different pharmacological classes, including psychomotor stimulants, opioids, barbiturates and 
benzodiazepines are investigated to develop an understanding of how drug-seeking becomes strong and 
persistent over time and how it might be weakened by pharmacologic or behavioral means. Laboratory 
objectives are pursued using a variety of experimental procedures, including ( 1 ) assessing the reinforcing 
effects of drugs of abuse using intravenous self-administration procedures, (2) quantifying then- 
behavioral effects using schedules of food presentation or electric shock delivery or postponement as 
baselines. (3) determining their effects as discriminative stimuli using two-lever and three-lever choice 
situations, and (4) determining the physiological effects of drugs of abuse, including cardiovascular 
toxicity and lethality. 

The Behavioral Pharmacology and Genetics Laboratory also conducts behavioral, pharmacological and 



67 



biochemical studies using genetically-defined animal models to investigate genetic factors underlying 
individual differences in response to the acute and chronic administration of drugs of abuse. The overall 
goal is to develop and apply behavioral pharmacogenetic approaches to assess and differentiate genetic 
and environmental factors underlying individual differences in response to drugs. Additional efforts are 
aimed at identifying, at the biochemical level, gene products which are involved in the acute, reinforcing 
and toxic effects of drugs of abuse, as well as the biochemical mechanisms underlying changes associated 
with the long-term administration of these drugs. In general, we are characterizing genetic factors 
associated with drug abuse and integrating this information with the results from the studies 
characterizing the environmental and behavioral factors associated with drug abuse. 



Summary of Ongoing Research 

A. Control of Behavior By Drug Injection 

Drugs serve as positive reinforcers to maintain and strengthen behavior leading to their administration 
and can control behavior through their ability to function as discriminative stimuli. In many situations, 
drugs of abuse probably function through multiple mechanisms to persistency sustain long sequences of 
drug seeking behavior that are very resistant to extinction. Schedule-controlled performances provide a 
meaningful way to analyze these long sequences of drug-seeking behavior in the same way as operant 
behavior maintained by other events such as food or shock. Using a variety of self-administration 
procedures, ongoing experiments are being conducted to evaluate behavior maintained by drugs and the 
ability of pharmacological treatments, such as antagonist administration or the development of 
dependence, to modify drug self-administration behavior and/or food-maintained behavior. 

One series of experiments involved the attempts at pharmacological modification of cocaine self- 
administration. In cardiovascular studies in squirrel monkeys (see project summary) we found that 
treatment with calcium channel blockers such as nimodipine will reverse or prevent cardiovascular 
changes produced by cocaine. However, presession treatments with calcium channel antagonists that 
were sufficient to reverse the effects of cocaine were without effect on cocaine-maintained behavior. In 
another series of experiments, the effects of administration of serotonergic drugs on the rate of 
responding by squirrel monkeys self-administering cocaine or responding for food pellets under fixed- 
ratio schedules of reinforcement are being conducted. In these studies, the effects of acute pretreatments 
of several 5HT2 antagonists will be evaluated in squirrel monkeys responding for cocaine. The direct 
effect of these pretreatments to disrupt ongoing food- maintained behavior will be assessed in the same 
subjects. 

Nicotine dependence is thought to be a a major obstacle to smoking cessation in man. Sertraline, a 
serotonergic uptake inhibitor that is effective as an antidepressant, is being evaluated for its usefulness as 
a potential treatment to facilitate smoking cessation. An ongoing project in the laboratory was designed 
to assess sertraline's effects in an animal model of nicotine dependence, nicotine self-injecting monkeys. 
Monkeys were initially trained to self-administer 10 to 30 «g/kg, i.v. cocaine under a fixed ratio 30, time- 
out 5 minute schedule of reinforcement. When the pattern of responding for cocaine was stable, a dose 
of nicotine (30 *g/kg, i.v.) that has been previously shown to maintain maximal rates of self-injection 
was substituted for cocaine. A dose-response curve for nicotine (0, 10, 30, 56, 100 «g/kg, i.v.) was then 
generated by substituting a series of nicotine doses for a period of 3 days. The second phase of the study 
involves evaluating the effects of sertraline pretreatments on self-administration behavior maintained by 
30 *g/kg nicotine. A series of sertraline doses or vehicle (0, 3,6, 12 mg/kg) will be administered by 
gavage prior to administration of each of three doses of nicotine or vehicle (0, 10, 30, 100 *g/kg, i.v.). 
As a control for non-specific effects of sertraline, nicotine or sertaline-nicotine combinations on food- 
reinforced responding, the effects of these drugs are being evaluated in squirrel monkeys. These 
monkeys are trained under the same fixed-ratio 30, time-out 5 minute schedule of reinforcement used in 
the nicotine self-injection study, except that food pellets are delivered instead of drug injections. A 



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nicotine dose response curve was generated by administering nicotine doses (0.1, 0.3 and 1.0 mg/kg, i.m.) 
prior to the session. As above, these doses of nicotine and vehicle will be evaluated in combination with 
sertraline. 

In addition to differences in pharmacological efficacy of drugs to control or modify behavior, it is clear 
that behavioral and environmental factors play an important role in the the control that even highly 
efficacious drugs exert on behavior. The focus of experiments in the newly established rhesus self- 
administration lab are to study the pharmacological, behavioral, and environmental variables involved in 
initiating and maintaining drug self-administration. Certain drugs, such as cocaine and other 
psychomotor stimulants generally function effectively as reinforcers under a variety of conditions. Other 
drugs such as benzodiazepines, some opioids and caffeine, however, have been studied only under 
relatively limited conditions, and generally maintain low levels of responding. The ability of these 
compounds to maintain self-injection behavior can be modified by a number of environmental and 
behavioral factors, such as stimuli associated with their administration, schedules of reinforcement, 
access conditions, concurrent availability of other drug and non-drug reinforcers (i.e., food), behavioral 
history and pharmacological manipulations such as antagonist administration or the development of 
tolerance/dependence. 

Parallel comparative studies in squirrel monkeys and humans in which subjects are given the opportunity 
to self-administer comparable doses of cocaine, morphine, nicotine and other drugs under similar 
behavioral schedules and experimental conditions provide a means to assess the generality of biological 
variables influencing drug self-administration. These studies allow an opportunity to evaluate the role of 
environmental variables and the role of conditioning in human drug taking behavior and whether those 
roles differ from the roles of those variables in animal models of drug taking. We have previously shown 
that second-order schedules of drug self-administration, where drug is injected only at the end of each 
daily session, can support a large amount of behavior in both humans and squirrel monkeys. In addition, 
in the human studies we have shown that with these schedules, low doses of morphine can support self- 
administration even though the subjects report no subjective effects of these drug doses. This result 
indicates that subjective report of a drug effect may not be an important factor in the positive reinforcing 
effects of drugs of abuse. To further substantiate these findings, we a currently repeating these 
experiments. In addition, we are also testing subjects who administer low morphine doses with the 
opioid antagonist naltrexone to determine whether any physiological sign of precipitated withdrawal can 
be observed. Subjective reports will also be taken during the withdrawal test to determine whether 
withdrawal may also occur independent of subjective report, or whether there is a closer match between 
(he subjective reports and withdrawal than between self-administration and subjective reports. 



B. Effects of Drugs on Schedule-Controlled Behavior of Experimental Animals 

General information on the behavioral pharmacology of a drug in a pertinent species is necessary to 
evaluate quantitatively how the drug functions as a reinforcer or a punisher as well as to establish a 
profile of behavioral effects. Schedules of food presentation with both fixed-interval and fixed-ratio 
components have been used most frequency in this type of study since they generate a wide range of 
rates and patterns of responding within a session and provide stable, long-term baselines for chronic 
studies in individual animals. The present project involves the assessment of both the acute and chronic 
effects of a variety of drugs on schedule-controlled behavior. 

We have recendy shown that while the calcium channel antagonists are effective in antagonizing the 
cardiovascular effects of cocaine, they do not significantly modify the direct behavioral effects of 
cocaine. This includes both the rate-increasing and rate decreasing effects of cocaine. We have also 
completed a long-term study designed to determine whether tolerance to either the rate-increasing or rate- 
decreasing effects of cocaine could be observed using a second-order schedule where the animals were 
allowed as long a period of time as needed to complete the schedule requirements. Under these 



69 



conditions, we did not observe any tolerance development to the effects of cocaine. 

We have previously shown that the opioid antagonist naltrexone produces clear behavioral sensitivity 
when its effects are determined on schedule-controlled behavior in rats. This phenomenon appears to be 
related to conditioning process, as it is long-lasting and undergoes extinction. These results indicate that 
opioid antagonist enhanced sensitivity may be an important factor in compliance in opioid abuse 
treatment with naltrexone. We have recendy shown that this enhanced sensitivity is pharmacologically 
specific, with cross-sensitivity occurring only to pure opioid antagonists. Further, we have also 
demonstrated that enhanced sensitivity occurs to salivation produced by high doses of naltrexone. This 
result is important as it allows us to study the phenomenon of enhanced sensitivity independently of 
schedule-controlled behavior. We have therefore studied receptor binding in these animals that show 
enhanced sensitivity with the salivation measure. In collaboration with Dr. Su in the Neuroscience 
branch we have shown that specific changes occur in opioid receptor binding. In addition, as opioid 
antagonist also interacts with GABA, we have been studying GABA binding. To date, those results have 
been inconclusive. 

Behaviorally active drugs can also serve as discriminative stimuli to guide behavioral choice. Ongoing 
drug discrimination projects in the laboratory using cocaine, opioids, caffeine and benzodiazepines 
agonists have helped to define and characterize the spectrum of behavioral effects produced by the drug, 
to compare a range of other compounds to characterize the relative potency and efficacy to produce drug- 
like effects, and to evaluate the drug's mechanisms of action at the receptor level. For example, we are 
currently studying the discriminative stimulus properties of the mu agonist morphine and the kappa 
agonist ethylketocyclazocine. In particular, we are interested to see if calcium channel antagonists, 
which can block some of the physiological effects of opioids, might also antagonize the discriminative 
stimulus properties of these drugs. Similar studies are also underway in human subjects under the 
direction of Dr. Vaupel from the Neuroscience Branch. In addition, we are also studying the effects of 
deprenyl, a monoamine oxidase inhibitor, to study the role of these processes in the discriminative 
stimulus properties of psychomotor stimulants. 

Since most human drug taking behavior involves chronic long-term use of an illicit drug or non-medical 
abuse of a prescribed medication, the consequences of chronic administration of drugs on schedule- 
controlled behavior and the discriminative functions of drugs are being evaluated. Although the 
development of tolerance and dependence appears to be related to pharmacological factors, such as 
chronic dose, period of exposure, conditions of administration (continuous vs. intermittent), tolerance can 
also be modified by environmental factors, such as the requirements of the task, presentation of 
conditioned stimuli, and the relationship between the task and the drug exposure. Manipulation of these 
environmental factors can result in the attenuation or elimination of tolerance even though high doses of 
drugs are being administered chronically. 

Behavioral factors have been shown to strongly influence the effects of chronic drug administration, often 
resulting in differential tolerance development. It is clear that tolerance can develop to the behavioral 
effects of benzodiazepines as a result of pharmacological exposure, but several studies have suggested 
that behavioral experience may only augment the rate of tolerance development. Recently, however, the 
demonstration of differential or contingent tolerance has been shown to be dependent upon the dose of 
the chronic drug, and the degree of training. Specifically, these studies have shown that the practice of a 
task under the influence of a drug using a PRE/POST injection strategy underlies the development of 
tolerance to the sedative effects of midazolam on food maintained behavior and to the effects of 
chlordiazepoxide on self-stimulation behavior. Ongoing studies in the lab have been designed to (1) 
assess the contributions of behavioral variables to the development of tolerance to the rate-decreasing 
effects of chlordiazepoxide (CDP) (i.e., contingent tolerance) and (2) determine the contributions of 
behavioral tolerance to CDP on sensitivity to acute administration of non-benzodiazepine compounds. 



70 



C. Cardiovascular Changes Induced by Cocaine. 

Several studies in the Behavioral Pharmacology and Genetics Laboratory are directed toward determining 
the effects of abused drugs on cardiovascular function. These studies are directed both at basic 
mechanisms and at potential treatment agents. We have found, for instance, that adrenergic mechanisms 
play an important role in the cardiovascular effects of cocaine. In studies in squirrel monkeys we have 
shown the alpha- 1 adrenergic mechanisms are important to cocaine's pressor effect, while both beta-1 and 
beta-2 adrenergic mechanisms are important to cocaine's tachycardiac effect. In particular, the alpha- 1 
antagonist prazocin was particularly effective in antagonizing cocaine's effects on cardiovascular function 
in squirrel monkeys. In parallel studies in rats, we have shown that prazocin can also antagonize the 
lethal effects of cocaine. Together, these results suggest that prazocin may be a particularly useful agent 
in the treatment of cocaine related medical emergencies. These studies have also shown that, contrary to 
previous finding, beta adrenergic antagonists should not be used in treatment. In rats, the beta antagonist 
propranolol will exacerbate cocaine-induced lethality. In squirrel monkeys, while propranolol will 
antagonize the tachycardiac effect of cocaine, it will potentiate the cocaine's pressor effect. Thus, beta 
antagonists should be avoided in treatment. 

An important aspect of this last year's research effort has been the development of effective techniques 
for studying cardiovascular function in conscious, freely-moving rats. These studies using conscious rats 
indicate that cocaine increases blood pressure and heart rate similar to its effects in squirrel monkeys. 
Further, a single injection of cocaine produces rapid sensitization to the pressor effects of its subsequent 
injections administered at 24 hr intervals. The cardiovascular effects of cocaine in rats are completely 
antagonized by noncompetitive or mixed type autonomic ganglionic blockers, while these effects are 
partially antagonized by the competitive ganglionic blockers. Cocaine also potentiates the peripheral 
cardiovascular effects of norepinephrine and inhibits the effects of tyramine, however, these effects occur 
at doses that are 10 times larger than those doses of cocaine alone required to produce cardiovascular 
effects. Thus, these results provide substantial evidence that the cardiovascular effects of cocaine in 
conscious rats are mainly centrally mediated. In agreement with this finding, we have recently shown 
that cocaine methiodide, a quaternary derivative of cocaine which should not cross the blood brain barrier 
does not affect cardiovascular function in conscious squirrel monkeys. Cocaine methiodide does 
potentiate the effects of exogenously administered norepinephrine, suggesting that it does have actions in 
the periphery. In anesthetized animals, some effects of cocaine methiodide are observed, although they 
are small and transient. Thus, our recent results from both rats and squirrel monkeys confirm that there is 
an important central component to the cardiovascular effects of cocaine. 

Recent studies in squirrel monkeys have also been directed toward comparing the effects of 
methamphetamine to cocaine on cardiovascular function. While the effects of methamphetamine appear 
to be mediated through the same noradrenergic processes as cocaine, an important aspect of 
methamphetamine s cardiovascular effects are also mediated through dopaminergic processes. This result 
suggests that cocaine and methamphetamine may act through different systems to affect cardiovascular 
function. 



D. Genetic Factors in Response to Chronic Drug Treatment 

In spite of the widespread recognition that there are strong individual differences in liability for drug 
abuse, relatively few studies designed to elucidate genetic factors associated with chronic drug use have 
been conducted. We are conducting a series of studies designed to evaluate pharmacogenetic differences 
in response to the chronic administration of drugs of abuse (cocaine and opioids) among genetically 
defined strains of mice and rats. The use of a pharmacogenetic approach not only facilitates the 
understanding of individual differences in response to chronic drug administration, it also provides a tool 
for understanding the biochemical mechanisms underlying these responses. We have investigated 
changes in susceptibility to cocaine-induced seizures following the long-term administration of cocaine 



71 



among inbred mouse strains. These studies revealed that genetic factors mediate not only acute 
sensitivity to the convulsant properties of cocaine but also changes in seizure susceptibility known to 
occur upon repeated administration of cocaine. The repeated administration of cocaine resulted in the 
development of either sensitization or tolerance to the convulsant effects of cocaine depending on the 
genetic background of the individual being examined. Having identified genotypes that are qualitatively 
different in their response to chronic cocaine, we are now using these animal models to address questions 
relating to the mechanisms underlying the effects of chronic cocaine. 

Cocaine exerts its effects through a number of biochemical systems. The convulsant and epileptogenic 
properties of cocaine are thought to be related, at least in part, to its local anesthetic effects. In a series of 
follow-up studies, we found that genetic factors also mediate sensitivity to the convulsant properties of 
the pure local anesthetic, lidocaine, and that there are genetic differences in response to the repeated 
administration of lidocaine, that is, depending on the genotype being, examined chronic lidocaine 
produced either an increased or a decreased sensitivity to cocaine-induced seizures. Furthermore, the 
chronic administration of subconvulsant doses of cocaine results in the development of cross-sensitization 
or cross-tolerance to the convulsant effects of lidocaine, again with the ultimate outcome being 
dependent upon the genetic background of the genotype being examined. Thus, the local anesthetic 
properties of cocaine may account for some of the individual differences in the effects of chronic cocaine 
administration. 

We have recently expanded our initial chrome cocaine studies to evaluate the ability of carbamazepine to 
modulate the convulsant and epileptogenic effects of cocaine. The primary site of action of 
carbamazepine is at voltage dependent sodium channels. These ion channels are known to be the site of 
action for local anesthetics such as cocaine and lidocaine. In that carbamazepine has been suggested as 
having some clinical use for the treatment of cocaine addiction, these studies are providing not only more 
information on the mechanisms underlying cocaine's effects, but also preclinical data on possible toxic 
side effects associated with the use of carbamazepine as a treatment for chronic cocaine use. Our results, 
to date, indicate that chronic, dietary carbamazepine attenuates, but does not completely inhibit, the 
cocaine-kindling process in a genotype-specific manner. Chronic carbamazepine also lowers the 
threshold for acute cocaine-induced seizures in some, but not all of the genotypes examined. This effect 
last for at least 3 days after the cessation of carbamazepine treatment, suggesting that some long-lasting 
biochemical change may have occurred. A finding of relevance to our earlier studies showing that 
chronic cocaine or lidocaine treatment resulted in sensitization or tolerance depending on the genotype 
being examined is the observation that chronic carbamazepine affected the development of tolerance, but 
not sensitization to the convulsant properties of cocaine. These studies have also shown that the regimen 
of administration of these drugs may have important implications in that a chronic period of 
carbamazepine administration is required for the attenuation of cocaine kindling and cocaine- induced 
seizures. These studies also revealed that the concurrent administration of carbamazepine and cocaine 
has genotype-specific lethal effects. Interestingly, the same data set suggests that while in one genotype 
the combination of the two drugs is lethal, in another mouse strain carbamazepine may serve attenuate 
the anorectic effects of cocaine. The mouse strains identified in the studies discussed above will serve as 
useful animal models for elucidating the mechanisms underlying some of the effects of chronic cocaine 
and/or carbamazepine. 

We have begun to examine changes in ion channel function and binding parameters associated with 
chronic drug administration. Preliminary results suggest that cocaine kindling may be associated with a 
decrease in the ability of the inhibitory neurotransmitter, GABA, to stimulate the uptake of chloride ions 
into neurons. In addition to continuing to examine these changes in GABAergic function, we have been 
developing techniques for assessment of other ligand- and voltage-gated ion channels, notably voltage- 
dependent sodium channels. While it is generally recognized that cocaine has major effects on these 
sodium channels, very few investigations have addressed the involvement of sodium channels in 
cocaine's actions. It is anticipated that these techniques will be useful for understanding the convulsant 
and epileptogenic properties of cocaine and should facilitate a greater understanding of the biochemical 
mechanisms underlying the effects of cocaine and the interactions between cocaine and carbamazepine. 



72 



Our studies are also being expanded to include assessment of biochemical changes resulting from the 
chronic administration of opioids and opioid antagonists. To date, we have identified an inbred mouse 
strain that differs from other strains we have examined in the number of neuronal mu opioid receptors 
and are correlating this difference with apparent differences in behavioral responses to opioid treatment. 
Similar biochemical assays are currently being conducted on brain tissue from rats chronically treated 
with an opioid antagonist. 



E. Pharmacogentics: Acute Response to Drug Administration 

The behavioral genetics model investigates individual differences in behavior as a composite of unique 
individual biology in context of its historical and present environment. To this end. behavior genetics 
analysis of individual differences in response to drugs of abuse such as alcohol and nicotine have proven 
useful in establishing heritable factors in acute response to ethanol and in determining traits with 
common genetic backgrounds. The ability to determine what physiological effects of ethanol are 
inherited independently has led to important molecular genetic discoveries and may lead to possible 
pharmacological interventions in the treatment of alcoholism. In addition to the work done with ethanol, 
previous studies investigating acute behavioral responses to opioids have demonstrated genotype to be an 
important factor. 

The use of inbred strain panels to estimate genetic correlations across ethanol-related phenotypes has 
proven useful in establishing heritable factors in acute response to ethanol and in determining traits with 
common genetic backgrounds. In addition to the work done with ethanol, previous studies investigating 
acute behavioral responses to opioids have demonstrated genotype to be an important factor. Initial 
studies in our laboratory suggest that the genetic rank order sensitivity among a number of opioid-related 
phenotypes such as analgesia, activity, hypothermia, muscle rigidity and receptor density do not 
consistently covary across genotype. Although opioid-related responses have been shown to have a 
heritable component, relatively few phenotypes have been investigated in a manner designed to 
determine estimates of phenotypic covariation. Determination of phenotypes with common or 
independent mechanisms is important for the development of compounds with therapeutic value 
independent of addiction liability and unwanted side-effects in addition a possible treatment for opioid 
addiction. The purpose of these studies is to determine the acute sensitivity of inbred strains and 
selected mouse lines for opioid-induced hypothermia, straub tail, analgesia and locomotion. The degree 
of physiological and behavioral covariance across genotype may indicate which genetic components are 
inherited in susceptibility to opioid-induced toxicities. In addition, these data would contribute 
significantly to ongoing investigations of genetic differences in opioid-reinforced behavior. 

In collaboration with Dr. Uhl of the Neuroscience Branch we are also evaluating behaviorally the acute 
response to opioids in transgenic mice which are over-expressing the endogenous opioid enkephalin. To 
date, the level of expression in brain has been insufficient to produce any significant behavioral changes. 
Additional mice have been recently bred which should have higher level of expression and biochemical 
assessments of genetic differences in opioid receptor binding parameters. 

The use of pbarmacogenetic methods is also useful in elucidating the biochemical mechanisms associated 
with drug responses. We are currently assessing the influence of ligand affinity for and receptor density 
of opiate and serotonergic receptor subtypes on opiate-induced analgesia, hypothermia and locomotion. 
We have also used pbarmacogenetic correlations to give us some information concerning the biochemical 
mechanisms mediating toxic responses to acute injections of cocaine. Using pharmacogenetic methods, 
we have shown clear genetic differences in sensitivity to cocaine-induced seizures and lethality across a 
variety of genetic strains of mice. Further, using a Classical Mendelian Analysis, we have data 
suggesting that a single gene associated with serotonergic 5HT2 receptors appears to mediate sensitivity 
to seizures. Using a different research strategy, pharmacological correlations, we have shown that the 



73 



binding of cocaine and related compounds to serotonin transporters appears to have a primary influence 
on seizures, while dopamine transporters has the greatest influence on lethal responses to cocaine. Drug 
binding to muscarinic and sigma receptors appears to attenuate both toxic responses. Related to these 
receptor binding studies, since polyamines appear to modulate transport systems for ions and 
neurotransmitters, and since cocaine produces its seizurgenic and lethal effects primarily via binding to 
serotonin and dopamine transporters, we intend to determine the effects of polyamines on cocaine- 
induced seizures and lethality. 

We are currently assessing the potential efficacy of several pharmacotherapeutic strategies on cocaine- 
induced toxicity, including sigma related drugs and antidepressants. We have observed a stereospecific 
effect of the sigma compound SKP10047 on cocaine-induced seizures. Since stereospecific effects of 
sigma compounds on behavioral phenomena have rarely been shown, we intend to assess the effects of 
several sigma ligands on measures of cocaine toxicity. It is hoped that the results may suggest antagonist 
and agonist properties of several sigma-related compounds. In addition, since cocaine produces its 
seizurgenic and lethal effects primarily via binding to serotonin and dopamine transporters, and since 
many antidepressant might be used clinically by potential cocaine users, we intend to determine the 
effects of various antidepressants on cocaine-induced toxicity. 

Finally, we have begun a project for the determination of the seizurgenic or lethal effects of cocethylene 
and several cocaine metabolites and congeners. Several such compounds have been shown in the blood 
or urine of cocaine-related emergency room cases. This project will assess whether these compounds 
might produce toxic effects by the same receptor mechanisms by which cocaine produces these effects. 
Thus, both biochemical and behavioral assessments of the potencies of these compounds will be 
performed. The results will be compared to our models of cocaine-induced seizures and lethality. 



F. Pharmacogenetic Factors in Drug Reinforced Behavior 

Previous studies in our laboratory have provided evidence to indicate an important genetic component in 
the acquisition of operant ethanol-reinforced behavior. Current research activities are aimed at 
determining the role of genotype in operant opioid-reinforced behavior, these studies include 
investigating the genetic and environmental components important not only in the acquisition but the 
maintenance and extinction of drug-reinforced behavior as well. The goal of our opioid drug self- 
administration project is to utilize recombinant inbred strains to investigate the role of opioid receptor 
subtypes and co-segregating opioid-related phenotypes in opioid self-administration. Opioid related 
phenotypes such as analgesia, hypothermia, activation, withdrawal and opioid-reinforced behavior, will 
be investigated in the parental strains C57BL/6 and BALB/C mice along with the CxBk subline. The 
CxBk mice are known to be deficient in mu-opioid binding sites and to be less sensitive than the C57BL 
strain to the analgesic and locomotor stimulatory effects of opioids. These data along with the use of 
several other CxB sublines will allow for the generation of phenotypic strain distribution patterns for in 
vitro and in vivo correlates of opioid reinforced behavior. Importantly, the use of the CxBk subline 
allows for the direct testing of known genetically inherited biochemical differences in the predisposition 
to opioid addiction. Thus far, several important findings have been made using this approach. Mu 
receptor populations may not account for individual variability in the acquisition of opioid-reinforced 
behavior following extensive training procedures. Inbred strains that varied significantly in mu receptor 
populations did not differ in the acquisition of etonitazene self-administration. Importantly, variation in 
mu receptor population may contribute to the extent to which an individual may self-administer opioids 
or the degree to which the behavior is maintained in the absence of drug availability. Individual 
variability may put some individuals at risk for acute opioid-induced toxicities during opioid self- 
administration. For example, C57BL/6 and CxBh mice do not differ in the respiratory depressant effects 
of etonitazene, yet drug intake to C57BL/6 mice is approximately four times the amount of CxBh mice 
during self-administration sessions. Collaborative efforts within the lab and in the Neuroscience Branch 
have begun to determine which underlying biochemical substrates may account for individual differences 



74 



seen in the propensity to exhibit opioid-reinforced behavior. 

In addition to investigating the genetic factors in acute sensitivity to opioids, current studies are also 
accessing the extent to which genetic factors affect conditioned place preference (CPP). The CPP 
paradigm measures the ability of a drug to engender conditioned reinforcement to previously neutral 
stimuli. The purpose of these studies are two-fold. First, these studies will determine if similar genetic 
differences are found in the ability of a drug to engender CPP compared to the propensity to exhibit 
operant self-administration. Similar genetic correlations across inbred strains would support common 
mechanism! s) responsible for maintaining operant self-administration and CPP. In addition, these studies 
are important for determining the reliability of the CPP paradigm to predict drug-reinforced behavior. 
Second, the genetic rank order for the ability of opioids to establish CPP will be correlated with the acute 
response to other opioid-related phenotypes in order to determine the degree of genetic covariation. 



75 



Articles Published or In Press 

Elmer. G.I. and George, F.R. Role of prostaglandin synthetase in the rate depressant effects and narcosis 
caused by ethanol. Journal of Pharmacology and Experimental Therapeutics , in press, 1990. 

Elmer, G.I.. Meisch, R.A., Goldberg, S.R. and George, F.R. Ethanol self-administration in long sleep 
and short sleep mice: Evidence for genetic independence of neurosensitivity and reinforcement. Journal 
of Pharmacology and Experimental Therapeutics . 254(3), 1054-1062, 1990. 

Gallager, D.W., R.J. Marley andT. D. Hernandez. Biochemical and electrophysiological mechanisms 
underlying benzodiazepine tolerance and dependence, in The Biological Basis of Drug Tolerance and 
Dependence , ed by J. Pratt, Academic Press, in press, 1990. 

George. F.R.. Ritz. MR., and Elmer. G.I. The role of genetics in drug dependence. In: The Biological 
Basis of Drug Tolerance and Dependence . Pratt. J. (ed). Academic Press. New York, in press. 

George, F.R., Elmer. G.I., Meisch. R.A. and Goldberg, S.R. Orally delivered cocaine functions as a 
positive reinibrcer in C57BL/6J mice. Pharmacology Biochemistry and Behavior , in press. 1990. 

George, F.R. and Ritz. M.C. Cocaine produces locomotor stimulation in SS but not LS mice: 
Relationship to dopaminergic function. Psvchopharmacologv . 101, 18-22, 1990. 

George, F.R.. Ritz. M.C. and Meisch. R.A. Ethanol produces similar fixed-ratio rate-depressant effects 
in ALKO AA and ANA rats. Advances in Alcohol and Substance Abuse . 9, 31-42, 1990. 

George, F.R., Porrino, L.J., Ritz, M.C. and Goldberg, S.R. Inbred rat strain comparisons indicate 
different sites of action for cocaine and amphetamine locomotor stimulant effects. Psvchopharmacologv , 
in press, 1990. 

George, F.R. and Ritz, M.C. Common mechanisms of reinforcement from alcohol and other drugs. In: 
Alcohol and Alcoholism , Suppl 1, in press. 1990. 

George. F.R. and Ritz. M.C. Cocaine-induced lethality is associated with interactions between dopamine 
transporters and muscarinic and sigma receptors. Journal of Pharmacology an d Experimental 
Therapeutics , in press. 1990. 

Goldberg, S. R.. Schindler, C. W., & Lamb, R. J. (1990) Second-order schedules and the analysis of 
human drug-seeking behavior. Drug Development Research , 20, 217-229. 

Griffiths, R.R.. Lamb, R.J., Sannerud C.A., Ator, N.A. and Brady, J.V. (1990) Self-injection of 
barbiturates, benzodiazepines and other sedative- anxiolytics in baboons. P^Ychoph annacologv. in press. 

Griffiths. R.R.. Evans. S.M.. Heishman, S.J.. Preston. K.L., Sannerud C.A., Wolf, B.A., and Woodson, 
P.P. Low-dose caffeine discrimination in humans. Journal of Pharmacology and Experimental 
Therapeutics . 252:970-978, 1990. 

Griffiths, R.R.. Evans. S.M., Heishman. S.J.. Preston, K.L., Sannerud, C.A., Wolf. B.A., and Woodson. 
P.P. Low-dose caffeine withdrawal. Journal of Pharmacology and Exp erimental Therapeutics. 25:1123- 
1132, 1990. 

Katz. J.L. and Goldberg, S.R. Second-order schedules of drug injection: Implications for understanding 
reinforcing effects of abused drugs. In: Advances in Substance Abuse. Be havioral and Biological 
Research Vol. 4. . N.K. Mello, Ed., Jessica Kingsley Publishers, Ltd, London, UK, in press. 



76 



Kuhar. M.J., Ritz, M.C. and Boja, J.W. The dopamine hypothesis of the reinforcing properties of 
cocaine. Trends in Neuroscience . in press. 1990. 

Lamb. R. J.. Preston. K. L.. Henningfield. J. E.. Schindler. C. W.. Meisch. R. L., Davis. R, Katz. J. L., & 
Goldberg, S. R. (1990) The reinforcing and subjective effects of morphine in post-addicts: A dose- 
response study. In press. 

Mansbach. R.S., Sannerud. C.A.. Griffiths. R.R., Balster, R.L., Harris, L.S. Intravenous self- 
administrauon of 4-methylaminorex in primates. Drug and Alcohol Dependence . 26:137-144,1990. 

Marley, R.J., C. Heninger, T.D. Hernandez and D.W. Gallager. Chronic administration of FG 7142 via. 
continuous i.e. v. infusion increases GABAergic function. Neuropharmacology, in press, 1990. 

Marley, R.J.. J. M. Witkin and S. R. Goldberg. Genetic factors influence changes in sensitivity to the 
convuisant properties of cocaine following chronic treatment. Rr?i p Re search, in press, 1990. 

Ritz. M.C, Boja. J.. George, F.R. and Kuhar. M.J. Cocaine binding sites related to drug self- 
administration. In: Problems of Drug Dependence 1989 . Louis S. Harris, Ed National Institute on Drug 
Abuse Research Monograph, 95, 239-246. 1990. 

Ritz. M.C, Cone, E.J. and Kuhar. M.J. Cocaine inhibition of ligand binding at dopamine, norepinephrine 
and serotonin transporters: A structure-activity study. Life Sciences . 46. 635-645. 1990. 

Ritz. M.C. Boja. J.. Carroll. F.I., Zaczak. R. and Kuhar. M.J.[ 3 H] WIN 35,065-2: A ligand for cocaine 
receptors in striatum. Journal of Neurochemistrv . 55, 1556-1562, 1990. 

Ritz, M.C Biochemical genetic differences in vulnerability to drug effects: Ls statistically significant 
always physiologically important and vice versa. Advances in Alcohol and Substance Abuse , in press, 
1990. 

Ritz, M.C, Kuhar, M.J. and George, F.R. Combined strategies from alcohol and drug research for 
determining the mechanisms of action of abused substances. In: Alcohol Abuse and A lcholism: Recent 
Advances . Van Thiel. D.H. and Tarter. R.E.. Eds.. Plenum Press, in press, 1990. 

Ritz. M.C, George, F.R. and Kuhar, M.J. Molecular mechanisms associated with cocaine effects: 
Possible relationships with effects of ethanol. In: Recent Developments in Alcohol ism: Alcohol and 
Cocaine: Clinical and Research Issues . Vol X, Gallant, DM., in press. 1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures are primarily associated with cocaine binding at 
serotonin transporters. Journal of Pharmacology and Experimental Therapeutics, in press. 1990. 

Sannerud. C.A., Ator. N.A., and Griffiths, R.R. (1990). Methocarbamol: evaluation of reinforcing and 
discriminative stimulus effects. Behavioural Pharmacology , in press. 

Sannerud. C.A., Ator, N.A., and Griffiths. R.R. (1990) Comparison of the discriminative stimulus effects 
of midazolam after intracranial and peripheral administration in the rat. Life Sciences (in press) 

Sannerud, C.A., Allen M., Cook. J.M., and Griffiths. R.R. (1990) Behavioral effects of benzodiazepine 
ligands in non-dependent, diazepam dependent and diazepam withdrawn baboons. European Journal of 
Pharmacology (in press). 



77 



Schindler. C. W.. Wu, X.-Z., Su. T.-S., Goldberg, S. R., & Katz. J. L. (1990) Enhanced sensitivity to 
behavioral effects of naltrexone in rats. Journal of Pharmacology and Experimental Therapeutics . 252. 8- 
14. 

Schindler. C. W., & Harvey, J. A. (1990) The use of classical conditioning procedures in behavioral 
pharmacology. Drug Development Research . 20, 169-187. 

Schindler, C. W„ White. M. F., & Goldberg, S. R. (1990) Effects of morphine, ethylketocyclazocine. N- 
allylnormetazocine and naloxone on locomotor activity in the rabbit. Psvchopharmacologv . 101. 172- 
177. 

Schindler. C. W.. TeUa. S. R., Wifkin, J. M., & Goldberg, S. R. (1990) Effects of cocaine alone and in 
combination with haloperidol and SCH 23390 on cardiovascular function in squirrel monkeys. Life 
Sciences , in press. 

Spear. D.J., Muntaner. C, Goldberg. S.R. and Katz. J.L. Methohexital and cocaine self-administration 
under fixed-ratio and second-order schedules. Pharmacology Biochemistry and Behavior , in press. 

Swedberg, M.D.B., Henningfield. J.E. and Goldberg, S.R. Nicotine dependency: animal studies. In: 
Nicotine Psvchopharmacologv: Molecular. Cellular and Behavioral Aspects . S. Wonnacott, (Ed.), 
M.A.H. Russell and l.P. Stolerman, Oxford University Press, Oxford, pp. 38-76, 1990. 

Telia, S. R., Schindler. C. W., & Goldberg, S. R. (1990) The role of central and autonomic neural 
mechanisms in the cardiovascular effects of cocaine in conscious squirrel monkeys. Journal of 
Pharmacology and Experimental Therapeutics . 252, 491-499. 

Telia, S. R., Schindler, C. W., & Goldberg, S. R. (1990) Rapid sensitization to the cardiovascular effects 
of cocaine in rats. European Journal of Pharmacology, in press. 

Telia. S. R., Schindler. C. W., & Goldberg, S. R. (1990) Cardiovascular effects of cocaine in squirrel 
monkeys. In P. V. Thadani (ed), Cardiovascular toxicity of eocen e: Underly ing mechanism;?, National 
Institute on Drug Abuse Research Monograph, in press. 

Thomas, D. A., Weiss, S. J., & Schindler. C. W. (1990) The effects of chlordiazepoxide and Ro 15-1788 
on preference for punished and unpunished response alternatives in rats. Psvchopharmacologv. 102. 333- 
338. 

Trouve, R., Nahas. G.G., Manger, W.M., Vinyard, C. and Goldberg, S.R. Interactions of nimodipine and 
cocaine on endogenous catecholamines in the squirrel monkey. Proc. Soc. Exp. Biol. & Med.. 193. 171- 
175. 1990. 

Wehner, J.M., B.J. Martin, R.J. Marley and J.I. Pounder. Behavioral studies of GABAergic responses in 
LS and SS mice: Are ethanol sensitivity and responses to GABAergic agents regulated by common 
mechanisms? In Initial Sensitivity to Ethanol . ed by R. A. Deitrick and A. A. Pawlowski, NIAAA Res. 
Monograph 20, pp. 345 - 380, 1990. 

Witkin, J.M. and Goldberg, S.R. Effects of cocaine on locomotor activity and schedule-controlled 
behaviors of inbred rat strains. Pharmacology Biochemistry and Behavior 37, 339-342, 1990. 

Witkin, J.M., Johnson, R.E., Jaffe, J.H., Goldberg, S.R., Grayson, N.A., Rice, K.C. and Katz, J.L. The 
partial opioid agonist, buprenorphine, protects against lethal effects of cocaine. Drug and Alcohol 
Dependence , in press. 



78 



Witkin. J. M.. Schindler, C. W., Telia. S. R.. & Goldberg, S. R. (1990) Interaction of haloperidol and 
SCH 23390 with cocaine and dopamine receptor subtype-selective agonists on schedule-controlled 
behavior of squirrel monkeys. Psychopharmacology. in press. 

Young, A.M., Sannerud, C.A., Steigerwald. E.S., Doty, M.D., Lipinski, W.J., and Tetrick, L.E. 
Tolerance to morphine stimulus control: Role of morphine maintenance dose. Psychopharmacology . 102. 
59-67. 1990. 



Abstracts Published or In Press 

Ator, N.A., Sannerud, C.A., and Griffiths. R.R. (1990) Abuse liability of benzodiazepine (BZ) receptor 
ligands. Proceeding from the 5th World Congress of Biological Psychiatry, Florence, Italy (in press). 

Colley, N.E., R.J. Marley & A.C. Collins. "Sleep-time and hypothermia mechanisms differ in LS and SS 
mouse lines. Abstracts of the Research Society on Alcoholism. 90. 1990. 

Elmer, G.I. and Goldberg, S.R. Genetic factors in the analgesic, stimulant, respiratory depressant and 
reinforcing properties of opioids. British Association for Psychopharmacology, in press. 1990. 

Elmer. G.I.. Pieper. J.OTJ.. Goldberg, S.R. and George, F.R. Genetic variation in opiate receptors and its 
relationship to opiate self-administration. Committee on Problems of Drug Dependence, in press 1990. 

George, F.R. and Ritz. M.C Cocaine-induced lethality: Mediation by dopamine uptake inhibition and 
direct action at muscarinic receptors. The FASEB Journal . #2775, 4, A745, 1990. 

George, F.R. and Ritz, M.C. Serotonin receptor densities appear to influence acquisition of ethanol- 
reinforced behavior. Alcoholism: Clinical and Experimental Research . 15, in press, 1990. 

Goldberg, S. R., Telia, S. R., & Schindler, C. W. (1990) Antagonism of cocaine-induced pressor 
response but not acute lethality by calcium channel entry blockers in rats. The FASEB Journal, 4, A744. 

Goldberg, S. R.. Telia. S. R., & Schindler, C. W. (1990) Adrenergic mechanisms in the cardiovascular 
effects of cocaine in squirrel monkeys. FASEB Journal, in press. 

Goldberg, S.R. & R. J. Marley. A pharmacogenetic approach towards an understanding of drug abuse. 
Abstracts of the British Association for Psychopharmacology, 1990. 

Griffiths. R.R.. Sannerud. C.A.. Lamb, R.J.. Ator, N.A., and Brady, J.V. (1990). Self-injection of 
barbiturates, benzodiazepines, and other sedative-anxiolytics in baboons. In: Problems of Drue 
Dependence 1990 . Harris, L.S. (ed). NIDA Research Monograph No. 105. Washington, D.C.:U.S. 
Government Printing Office, 329. 

Korupolu, G. R.. Telia, S. R.. Schindler. C. W., & Goldberg, S. R. (1990) Modification of cocaine- 
induced cardiovascular effects, convulsions and lethality by adrenoceptor blocking agents in rats. FASEB 
Journal, in press. 

Marley, R.J., J.M. Witkin, & S.R. Goldberg. A pharmacogenetic evaluation of the cocaine-kindling 
process. Society for Neuroscience Abstracts, #1133, 1990. 

Marley, R.J., J. M. Witkin and S. R. Goldberg. Genetic differences in the development of cocaine- 
kindled seizures. NIDA Monograph: Problems of Drug Dependence, 1990, in press. 



79 



Ritz, M.C. and George, F.R. Cocaine-induced seizures: Mediation by specific CNS receptors (presented 
at the 1989 annual ACNP meeting). Neuropsvchopharmacologv . 1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures are initiated by serotonin uptake inhibition, but 
attenuated by direct action at sigma and muscarinic receptors. The FASEB Journal . #2776, 4, A745, 
1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures and lethality: Mediation by distinct central 
nervous system receptors. Problems of Drug dependence 1990 . Proceedings of the 52nd Annual 
Scientific Meeting, Committee of Problems of Drug Dependence, Inc. National Institute on Drug Abuse 
Research Monograph, in press, 1990. 

Ritz, M.C. and George, F.R. Cocaine toxicity: Distinct neurotransmitter systems are associated with 
seizures and death. Society for Neuroscience Abstracts . #242.8, 16, 581, 1990. 

Ritz, M.C. and George, F.R. Antidepressant drugs appear to enhance cocaine-induced toxicity (presented 
at the 1990 anual ACNP meeting ). Neuropsvcopharmacologv . in press, 1990. 

Sannerud. C.A., Kaminski. B.J. and Griffiths, R.R. (1990) Lack of self-injection behavior maintained by 
N-N-dimethylamphetamine in baboons. The FASEB Journal , in press. 

Sannerud. C.A., Ator. N.A.. Fischette, C.T., and Griffiths, R.R. (1990). Behavioral effects of 
landospirone in male baboons. Society for Neurosciences Abstracts 16, 1322. 

Sannerud. C.A. and Griffiths. R.R. (1990) Behavioral effects of chronic abecarnil administration in 
baboons. In: Problems of Drug Dependence 1990 . Harris, L.S. (ed). NJJDA Research Monograph No. 
105. Washington. D.C.:U.S. Government Printing Office, 328, in press 

Sannerud, C.A. and Griffiths, R.R. (1990) Assessment of benzodiazepine physical dependence in 
baboons. Pharmacology. Biochemistry and Behavior in press. 

Sannerud, C.A., Alastra, A.J.G., and Harger, P.L.(1990) Contingent tolerance to chlordiazepoxide in rats. 
Pharmacology. Biochemistry and Behavior , in press. 

Schindler, C. W., Telia. S. R.. Prada. J. A., & Goldberg, S. R. (1990) Failure of calcium channel entry 
blockers to antagonize the behavioral effects of cocaine in squirrel monkeys. The FASEB Journal . 4, 

A745. 

Schindler, C. W., Goldberg, S. R., & Katz, J. L. (1990) Pharmacological specificity of enhanced 
sensitivity to naltrexone in rats. Pharmacology Biochemistry and. Behavior . 36, 438. 

Schindler, C. W„ Wu. X.-Z.. Su, T.-P.. Thorndike. E. B., Goldberg, S. R., & Katz, J. L. (1990) 
Enhanced sensitivity to naltrexone in rats: Effects on salivation and opioid receptor binding. Society far 
Neuroscience Abstracts , 16, 1 1 92. 

Schindler. C. W ., Telia, S. R.. Katz, J. L., & Goldberg, S. R. (1990) Effects of cocaine and cocaine 
methiodide on cardiovascular function in squirrel monkeys. FASEB Journal , in press. 

Telia, S. R., Schindler, C. W., & Goldberg, S. R. (1990) Antagonism of cocaine-induced cardiovascular 
changes by calcium channel entry blockers in conscious squirrel monkeys. The FASEB Journal . 4, A744. 

Telia, S. R. Schindler. C. W., & Goldberg, S. R. (1990) A single intravenous bolus injection of cocaine 
enhances the pressor effects of subsequent injection in conscious rats. Circulation . 82, III-147. 



80 



Telia, S. R., Schindler, C. W., Konjpolu. G. R., & Goldberg, S. R. (1990) Variable degree of antagonism 
of cardiovascular effects of cocaine by ganglionic blockers in conscious rats. FASEB Journal , in press 

Wilkin, J.M., S.R. Goldberg & R.J. Marley. Lack of a genetic correlation between the convulsant and 
epdeptogenic effects of cocaine and lidocaine. Abstracts of Comm. Prob. Drug Dep. 52nd Ann. Sci. 
Mtg., 1990. 



81 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

Period Covered: January 1, 1990 to December 31, 1990 
Title of Project: Control of Behavior by Drug Injection 
Principal Investigators: 



Z01 DA 00001-06 BPL 



PL: 


S.R. Goldberg 


Chief 


Others: 


C.W. Schindler 


Research Psychologist 




J. A. Prada 


Research Psychologist 




C.A. Sannerud 


Staff Fellow 




J.-W. Zheng 


Visiting Associate 




S. Yasar 


Visiting Fellow 



BPGL. NIDA, ARC 

BPGL, NIDA, ARC 
BPGL, NIDA, ARC 
BPGL, NIDA. ARC 
BPGL, NIDA, ARC 
BPGL. NIDA. ARC 



Cooperating Units: None 
Lab/Branch: 



Behavioral Pharmacology and Genetics Laboratory 
Preclinical Pharmacology Branch 



Section: None 



Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, 
Baltimore, MD 21224 



Total Man Years: 



2.50 



Professional: 



2.00 



Check Appropriate Boxes: 

Human Subjects 

Minors 

Interviews 



Human Tissues 



Other: 0.50 



X Neither 



Summary of Work: 

Using self-administration procedures, several different types of experiments are being conducted The 
abuse liability of drugs are being assessed by comparing the rates and patterns of responding maintained 
by various drugs, including cocaine, nicotine and other psychomotor stimulants, benzodiazepines and 
other sedative/anxiolytics, morphine and other opioids. These studies will compare responding 
maintained under fixed-ratio, fixed-interval schedules and complex second-order schedules. The ability 
of pharmacological treatments and the development of tolerance/dependence to modify drug self- 
administration behavior and/or food maintained behavior is also being assessed. In addition to 
differences in pharmacological efficacy of drugs, it is clear that behavioral and environmental factors can 
modify the control that even highly efficacious drugs exert on behavior. The focus of experiments in the 
rhesus self-administration lab are to study the pharmacological, behavioral, and environmental variables 
involved in initiating and maintaining drug self-administration. Certain drugs, such as cocaine and other 
psychomotor stimulants generally function effectively as reinforcers under a variety of conditions. Other 
drugs such as benzodiazepines, some opioids, and caffeine, however, have been studied only under 
relatively limited conditions, and generally maintain low levels of responding. Parallel comparative 
studies in squirrel monkeys and humans in which subjects are given the opportunity to self-administer 
comparable doses of cocaine, morphine, nicotine and other drugs under similar behavioral schedules and 



82 



experimental conditions provide a means to assess the generality of biological variables influencing drug 
self-administration. We have previously shown under a second-order schedule of drug self- 
administration, low doses of morphine can support self-administration in humans even though the 
subjects report no subjective effects of these drug doses. This result indicates that subjective reports of a 
drug effect may not be an important factor in the positive reinforcing effects of drugs of abuse. To 
further substantiate these findings, we are currently repeating these experiments. In addition, we are also 
testing subjects who administer low morphine doses with the opioid antagonist naltrexone to determine 
whether any physiological signs of precipitated withdrawal can be observed. Subjective reports will also 
taken during the withdrawal test to determine whether withdrawal may also occur independent of 
subjective report, or whether there is a closer match between the subjective reports and withdrawal than 
between self-administration and subjective reports. 

Publications 

Goldberg, S. R., Schindler. C. W.. & Lamb. R. J. (1990) Second-order schedules and the analysis of 
human drug-seeking behavior. Drug Development Research, 20, 217-229. 

Griffiths, R.R.. Lamb, R.J.. Sannenid, C.A., Ator, N.A. and Brady, J.V. (1990) Self-injection of 
barbiturates, benzodiazepines and other sedative-anxiolytics in baboons. Psvchopharmacology. in press. 

Katz. J.L. and Goldberg, S.R. Second-order schedules of drug injection: Implications for understanding 
reinforcing effects of abused drugs. In: Advances in Substance Abuse. Behavioral and Biological 
Research Vol. 4. . N.K. Mello. Ed. Jessica Kingsley Publishers, Ltd., London, UK, in press. 

Lamb. R. J.. Preston. K. L.. Henningfield. J. E., Schindler, C. W., Meisch, R. L., Davis, E, Katz. J. L., & 
Goldberg, S. R. (1990) The reinforcing and subjective effects of morphine in post-addicts: A dose- 
response study, in press. 

Mansbach. R.S., Sannenid, C.A., Griffiths, R.R., Balster, R.L., Harris, L.S. Intravenous self- 
administration of 4-methylaminorex in primates. Drug and Alcohol Dependence . 26:137-144,1990. 

Sannenid, C.A., Ator, N.A., and Griffiths, R.R. (1990). Methocarbamol: evaluation of reinforcing and 
discriminative stimulus effects. Behavioural Pharmacology , in press. 

Spear. D.J.. Muntaner. C, Goldberg. S.R. and Katz, J.L. Methohexital and cocaine self-administration 
under fixed-ratio and second-order schedules. Pharmacology Biochemistry and Behavior , in press. 

Swedberg, M.D.B., Henningfield. J.E. and Goldberg, S.R. Nicotine dependency: animal studies. In: 
Nicotine Psvchopharmacology: Molecular. Cellular and Behavioral Aspects . S. Wonnacott, (Ed.), 
M.A.H. Russell and LP. Stolerman. Oxford University Press, Oxford, pp. 38-76. 1990. 

Abstracts 

Ator. N.A.. Sannenid, C.A., and Griffiths. R.R. (1990) Abuse liability of benzodiazepine (BZ) receptor 
ligands. Proceeding from the 5th World Congress of Biological Psychiatry, Florence, Italy (in press). 

Griffiths. R.R.. Sannenid, C.A.. Lamb. R.J., Ator, N.A., and Brady, J.V. (1990). Self-injection of 
barbiturates, benzodiazepines, and other sedative-anxiolytics in baboons. In: Problems of Drug 
Dependence 1990 . Harris. L.S. (ed). NIDA Research Monograph No. 105. Washington. D.C.:U.S. 
Government Printing Office, 329. 

Sannenid, C.A., Kaminski, B.J. and Griffiths, R.R. (1990) Lack of self-injection behavior maintained by 
N-N-dimethylamphetamine in baboons. The FASEB Journal, in press. 



83 



Schindler. C. W., TeUa. S. R., Prada, J. A., & Goldberg, S. R. (1990) Failure of calcium channel entry 
blockers to antagonize the behavioral effects of cocaine in squirrel monkeys. Ihe. FASEB Journal . 4, 

A745. 



84 



NOTICE OF INTRAMURAL RESEARCH PROJECT 



Z01 DA 00003-06 BPL 



Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Effects of Drags on Schedule-Controlled Behavior of Experimental Animals 



Principal Investigators: 



P.I. 


S.R. Goldberg 


Others: 


J.A. Prada 




C.W. Schindler 




C.A. Sannerad 




R.J. Marley 




J.-W. Zheng 




S. Yasar 



Cooperating Units: None 



Chief 

Research Psychologist 
Research Psychologist 
Staff Fellow 
Staff Fellow 
Visiting Associate 
Visiting Fellow 



BPGL, NIDA, ARC 



BPGL, 
BPGL, 
BPGL, 
BPGL, 
BPGL, 
BPGL. 



Lab/Branch: 



Section: None 



NIDA 
NJDA 
NIDA 
NIDA 
NIDA 
NIDA 



ARC 
ARC 
ARC 
ARC 
ARC 
ARC 



Behavioral Pharmacology and Genetics Laboratory 
Preclinical Pharmacology Branch 



Institute and Location: 

Addiction Research Center, National Institute on Drag Abuse, 
Baltimore. MD 21224 



Total Man Years: 



4.46 



Professional: 



.76 



Other: 1.70 



Check Appropriate Boxes: 

Human Subjects 

Minors 

Interviews 



Human Tissues 



X Neither 



Summary of Work: 

General information on the behavioral pharmacology of a drag in a pertinent species is necessary to 
evaluate quantitatively how the drag functions as a reinforcer or a punisher as well as to establish a 
profile of behavioral effects. Schedules of food presentation with both fixed-interval and fixed-ratio 
components have been used most frequendy used in this type of study since they generate a wide range of 
rates and patterns of responding within a session and provide stable, long-term baselines for chronic 
studies in individual animals. The present project involves the assessment of both the acute and chronic 
effects of a vanety of drags on schedule-controlled behavior. We have recendy shown while the calcium 
channel antagonists are effective in antagonizing the cardiovascular effects of cocaine, they do not 
significanUy modify the direct behavioral effects of cocaine. This includes both the rate-increasing and 
rate-decreasing effects of cocaine. We have also completed a long-term study designed to determine 
whether tolerance to either the rate-increasing or rate-decreasing effects of cocaine could be observed 
using a second-order schedule where the animals were allowed as long a period of time as needed to 
complete the schedule requirements. Under these conditions, we did not observe any tolerance 
development to the effects of cocaine. We have also recendy shown that the enhanced sensitivity which 



85 



occurs to opioid antagonists is pharmacologically specific, with cross-sensitivity occurring only to pure 
opioid antagonists. Further, we have demonstrated that this enhanced sensitivity occurs to salivation 
produced by high doses of naltrexone. This result is important as it allows us to study the phenomenon of 
enhanced sensitivity independently of schedule-controlled behavior. In collaboration with Dr. Su in the 
Neuroscience branch we have shown that specific changes occur in opioid receptor binding following 
opioid antagonist treatment. In addition to their direct effects on behavior, drugs of abuse can also 
function as discriminative stimuli. Most of these studies are being performed in rats and we are studying 
the discriminative stimulus effects of cocaine and other psychomotor stimulants, opioids and the 
benzodiazepines. These studies are designed to characterize the relative potency and efficacy of test 
drugs to produce drug-like effects, and to evaluate the drug's mechanism of action at the receptor level. 
For example, we are curremly conducting experiments to determine if calcium channel antagonists, 
which can block some of the physiological effects of opioids, might also antagonize the discriminative 
stimulus properties of these drugs. Similar studies are also underway in human subjects under the 
direction of Dr. Vaupel from the Neuroscience Branch. 

Publications 

Griffiths. R.R.. Evans. S.M., Heishman. S.J.. Preston. K.L., Sannerud. C.A., Wolf. B.A.. and Woodson, 
P.P. Low-dose caffeine discrimination in humans. Journal of Pharmacology and Experimental 
Therapeutics . 252:970-978, 1990. 

Griffiths. R.R., Evans. S.M., Heishman. S.J.. Preston, K.L., Sannerud. C.A.. Wolf, B.A.. and Woodson, 
P.P. Low-dose caffeine withdrawal. Journal of Pharmacology and Experimental Therapeutics . 25:1123- 
1132, 1990. 

Sannerud. C.A., Ator, N.A.. and Griffiths, R.R. (1990) Comparison of the discriminative stimulus effects 
of midazolam after intracranial and peripheral administration in the rat. Life Sciences (in press) 

Sannerud. C.A., Allen M., Cook, J.M., and Griffiths. R.R. (1990) Behavioral effects of benzodiazepine 
ligands in non-dependent, diazepam dependent and diazepam withdrawn baboons. European Journal of 
Pharmacology (in press). 

Schindler. C. W.. Wu. X.-Z.. Su, T.-S., Goldberg, S. R.. & Katz. J. L. (1990) Enhanced sensitivity to 
behavioral effects of naltrexone in rats. Journal of Pharmacology and Experimental Therapeutics . 252, 8- 
14. 

Schindler. C. W., & Harvey, J. A. (1990) The use of classical conditioning procedures in behavioral 
pharmacology. Drug Development Research, 20, 169-187. 

Schindler. C. W., White. M. F.. & Goldberg, S. R. (1990) Effects of morphine, ethylketocyclazocine. N- 
allylnormetazocine and naloxone on locomotor activity in the rabbit. Psvchopharmacologv . 101, 172- 
177. 

Thomas. D. A.. Weiss. S. J., & Schindler, C. W. (1990) The effects of chlordiazepoxide and Ro 15-1788 
on preference for punished and unpunished response alternatives in rats. Psvchopharmacologv , 102, 333- 
338. 

Witkin, J.M. and Goldberg, S.R. Effects of cocaine on locomotor activity and schedule-controlled 
behaviors of inbred rat strains. Pharmacology Biochemistry and Behavior 37, 339-342, 1990. 

Witkin, J. M., Schindler, C. W., Telia. S. R., & Goldberg, S. R. (1990) Interaction of haloperidol and 
SCH 23390 with cocaine and dopamine receptor subtype-selective agonists on schedule-controlled 
behavior of squirrel monkeys. Psvchopharmacologv . in press. 



86 



Young, A.M.. Sannerud. C.A., Steigenvald, E.S., Doty, M.D., Lipinski, W.J., and Tetrick. L.E. 
Tolerance to morphine stimulus control: Role of morphine maintenance dose. Psvchopharmacology . 
102:59-67. 1990. 



Abstracts 

Sannerud. C.A., Ator, N.A., Fischette, C.T., and Griffiths. R.R. (1990). Behavioral effects of 
tandospirone in male baboons. Society for Neurosciences Abstracts 16, 1322. 

Sannerud, C.A. and Griffiths. R.R. (1990) Behavioral effects of chronic abecamil administration in 
baboons. In: Problems of Drue Dependence 1990 . Harris, L.S. (ed). NIDA Research Monograph No. 
105. Washington. D.C.:U.S. Government Printing Office. 328, in press 

Sannerud, C.A. and Griffiths, R.R. (1990) Assessment of benzodiazepine physical dependence in 
baboons. Pharmacology. Biochemistry and Behavior (in press). 

Sannerud. C.A.. Alastra. A.J.G.. and Harger. P.L.(1990) Contingent tolerance to chlordiazepoxide in rats. 
Pharmacology. Biochemistry and Behavior , (in press). 

Schindler. C. W., Telia, S. R.. Prada, J. A., & Goldberg, S. R. (1990) Failure of calcium channel entry 
blockers to antagonize the behavioral effects of cocaine in squirrel monkeys. The FASEB Journal . 4, 

A745. 

Schindler, C. W., Goldberg, S. R., & Katz, J. L. (1990) Pharmacological specificity of enhanced 
sensitivity to naltrexone in rats. Pharmacology Biochemistry and Behavior . 36, 438. 

Schindler, C. W., Wu, X.-Z.. Su, T.-P., Thorndike, E. B., Goldberg, S. R., & Katz, J. L. (1990) 
Enhanced sensitivity to naltrexone in rats: Effects on salivation and opioid receptor binding. Society for 
Neuroscience Abstracts . 16, 1192. 



87 



NOTICE OF INTRAMURAL RESEARCH PROJECT 
Period Covered: January 1. 1990 to December 31, 1990 
Title of Project: Cardiovascular Changes Induced by Cocaine 
Principal Investigators: 

P.I.:. C.W. Schindler Research Psychologist 

Others: 



Z01 DA 00009-04 BPL 



S.R. Goldberg 
S.R. Telia 
G.R. Korupolu 
J.-W. Zheng 



Chief 
Guest Worker 
Visiting Fellow 
Visiting Associate 



BPGL, NIDA, ARC 

BPGL, NIDA. ARC 
BPGL. NIDA. ARC 
BPGL, NIDA, ARC 
BPGL, NDDA, ARC 



Cooperating Units: None 



Lab/Branch: 



Section: None 



Behavioral Pharmacology and Genetics Laboratory 
Preclinical Pharmacology Branch 



Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, 
Baltimore, MD 21224 



Total Man Years: 



2.86 



Professional: 



.86 



Other: 



Check Appropriate Boxes: 

Human Subjects Human Tissues X_ Neither 

Minors 

Interviews 

Summary of Work: 

The effects of cocaine and other psychomotor stimulants on a number of physiological parameters are 
being studied in conscious squirrel monkeys and rats. Recent studies in squirrel monkeys have indicated 
that alpha- 1 adrenergic mechanisms are importantly involved in the pressor effects of both cocaine and 
methamphetamine, while both beta-1 and beta-2 adrenergic mechanisms are important for the 
tachycardiac effect of both drugs. Based on these studies, the alpha- 1 antagonist prazocin would appear 
to be an ideal drug for the treatment of cardiovascular complications due to psychomotor stimulant abuse. 
Unlike with cocaine, dopaminergic mechanisms have been shown to be importantly involved in the 
cardiovascular effects of methamphetamine. In addition, recent studies have also indicated that, contrary 
to our previous findings, central mechanisms do appear to be significantly involved in the cardiovascular 
effects of cocaine in squirrel monkeys. The studies using conscious rats indicate that cocaine increases 
blood pressure and heart rate similar to its effects in squirrel monkeys. Further, a single injection of 
cocaine produces rapid sensitization to the pressor effects of its subsequent injections administered at 24 
hr intervals. The cardiovascular effects of cocaine in rats are completely antagonized by noncompetitive 
or mixed type autonomic ganglionic blockers, while these effects are partially antagonized by the 
competitive ganglionic blockers. Cocaine also potentiates the peripheral cardiovascular effects of 
norepinephrine and inhibits the effects of tyramine, however, these effects occur at doses that are 10 



88 



times larger than those doses of cocaine alone required to produce cardiovascular effects. Thus, these 
results provide substantial evidence that the cardiovascular effects of cocaine in conscious rats are mainly 
centrally medicated. Acute lethality studies indicate that various adrenergic agents modify acute cocaine 
intoxication. As with the cardiovascular effects in squirrel monkeys, prazocin was particularly effective 
against cocaine lethality. 

Publications 

Schindler, C. W„ Telia. S. R., Witkin, J. M., & Goldberg, S. R. (1990) Effects of cocaine alone and in 
combination with haloperidol and SCH 23390 on cardiovascular function in squirrel monkeys. Life 
Sciences , in press. 

Telia, S. R., Schindler, C. W., & Goldberg, S. R. (1990) The role of central and autonomic neural 
mechanisms in the cardiovascular effects of cocaine in conscious squirrel monkeys. Journal of 
Pharmacology and Experimental Therapeutics . 252, 491-499. 

Telia. S. R.. Schindler. C. W., & Goldberg, S. R. (1990) Rapid sensitization to the cardiovascular effects 
of cocaine in rats. European Journal of Pharmacology , in press. 

Telia. S. R., Schindler. C. W., & Goldberg, S. R. (1990) Cardiovascular effects of cocaine in squirrel 
monkeys. In P. V. Thadani (ed.). Cardiovascular toxicity of cocaine: Underlying mechanisms . National 
Institute on Drug Abuse Research Monograph, in press. 

Trouve, R., Nahas, G.G, Manger, W.M., Vinyard, C. and Goldberg, S.R. Interactions of nimodipine and 
cocaine on endogenous catecholamines in the squirrel monkey. Proc. Soc. Exp. Biol. & Med. . 193, 171- 
175, 1990. 

Witkin, J.M.. Johnson. RE., Jaffe, J.H.. Goldberg, S.R., Grayson, N.A., Rice. K.C. and Katz, J.L. The 
partial opioid agonist, buprenorphine, protects against lethal effects of cocaine. Drue and Alcohol 
Dependence , in press. 

Abstracts 

Goldberg, S. R., Telia, S. R., & Schindler, C. W. (1990) Antagonism of cocaine-induced pressor 
response but not acute lethality by calcium channel entry blockers in rats. The FASEB Journal . 4, A744. 

Goldberg, S. R., Telia, S. R., & Schindler, C. W. (1990) Adrenergic mechanisms in the cardiovascular 
effects of cocaine in squirrel monkeys. The FASEB Journal , in press. 

Korupolu. G. R.. Telia. S. R., Schindler, C. W., & Goldberg, S. R. (1990) Modification of cocaine- 
induced cardiovascular effects, convulsions and lethality by adrenoceptor blocking agents in rats. The 
FASEB Journal , in press. 

Schindler, C. W., Telia, S. R., Katz. J. L., & Goldberg, S. R. (1990) Effects of cocaine and cocaine 
methiodide on cardiovascular function in squirrel monkeys. The FASEB Journal , in press. 

Telia, S. R„ Schindler, C. W., & Goldberg, S. R. (1990) Antagonism of cocaine-induced cardiovascular 
changes by calcium channel entry blockers in conscious squirrel monkeys. The FASEB Journal . 4, A744. 

Telia, S. R. Schindler. C. W., & Goldberg, S. R. (1990) A single intravenous bolus injection of cocaine 
enhances the pressor effects of subsequent injection in conscious rats. Circulation. 82, III- 147. 



89 



Telia. S. R.. Schindler, C. W., Korupolu. G. R.. & Goldberg, S. R. (1990) Variable degree of antagonism 
of cardiovascular effects of cocaine by ganglionic blockers in conscious rats. The FASEB Journal , in 
press 



90 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00012-02-BPL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: Genetic Factors in Response to Chronic Drug Treatment 

Principal Investigators: 



P.I.: 


R.J. Marley 


Staff Fellow 


BPGL. NJX>A, ARC 


Others: 


S.R. Goldberg 


Chief 


BPGL. NJDA, ARC 




N.L. Goodman 


Research Pharmacologist 


BPGL. NJDA, ARC 



Cooperating Units: None 

Lab/Branch: Behavioral Pharmacology and Genetics Laboratory 

Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse. 
Baltimore, MD 21224 

Total Man Years: 1.36 Professional: 0.76 Other: 0.60 

Check Appropriate Boxes: 

Human Subjects Human Tissues X_ Neither 

Minors 

Interviews 

Summary of Work: 

The purpose of this project is to use a pharmacogenetic approach to evaluate individual differences in 
response to chronic administration of drugs of abuse or drugs proposed for the treatment of drug abuse. 
Genetic differences in response to the convulsant and epileptogenic effects of cocaine have been 
demonstrated by examining the development of increased susceptibility to the seizure inducing properties 
of cocaine following repeated administration of the drug (pharmacological kindling). Genetically distinct 
strains of mice that differ quantitatively and qualitatively in their response to chronic cocaine have been 
identified and are being employed to study the mechanisms underlying the effects of long-term cocaine 
use and to assess possible treatments for cocaine toxicity. We have demonstrated that individual 
differences in response to the epileptogenic effects of cocaine are associated with its local anesthetic 
properties. Evaluation of the ability of carbamazepine to modulate the convulsant and epileptogenic 
effects of cocaine have shown that chronic carbamazepine attenuates the development of increased 
sensitivity to cocaine- induced seizures in a genotype-specific manner. In addition to genetic difference, 
the regimen of carbamazepine administration appears to be important in determining its ability to 
attenuate cocaine's effects. These studies also revealed that chronic carbamazepine administration, in 
conjunction with chronic cocaine treatment, has lethal consequences in certain genotypes. Assessment of 
the biochemical mechanisms underlying the changes observed following chronic cocaine treatment are 
presently being conducted. Preliminary results suggest that cocaine kindling is associated with a 
downregulation of GABAergic function. Genetic differences in response to pharmacological kindling 



91 



with a benzodiazepine inverse agonist have also been demonstrated and are being compared with the 
differences observed for cocaine kindling. Assessments of biochemical changes associated with chronic 
opiate treatment are also being conducted. 



Publications 

Gallager, D.W., R.J. Marley and T. D. Hernandez. Biochemical and electrophysiological mechanisms 
underlying benzodiazepine tolerance and dependence, in The Biological Basis of Drug Tolerance and 
Dependence , ed by J. Pratt, Academic Press, in press. 1990. 

Marley, R.J., C. Heninger, T.D. Hernandez and D.W. Gallager. Chronic administration of FG 7142 via. 
continuous i.e. v. infusion increases GABAergic function. Neuropharmacology , in press, 1990. 

Marley, R.J.. J. M. Witkin and S. R. Goldberg. Genetic factors influence changes in sensitivity to the 
convulsant properties of cocaine following chronic treatment. Brain Res. , in press. 1990. 

Wehner, J.M.. B.J. Martin, R.J. Marley and J.I. Pounder. Behavioral studies of GABAergic responses in 
LS and SS mice: Are ethanol sensitivity and responses to GABAergic agents regulated by common 
mechanisms? In Initial Sensitivity to Ethanol . ed by R. A. Deitrick and A. A. Pawlowski, NIAAA Res. 
Monograph 20 . pp. 345 - 380. 1990. 

Abstracts 

Colley, N.E.. R.J. Marley & A.C. Collins. "Sleep-time and hypothermia mechanisms differ in LS and SS 
mouse lines. Abstracts of the Research Society on Alcoholism . 90, 1990. 

Goldberg, S.R. & R. J. Marley. A pharmacogenetic approach towards an understanding of drug abuse. 
Abstracts of the British Association for Psvchopharmacolog y. 1990. 

Marley, R.J., J.M. Witkin, & S.R. Goldberg. A pharmacogenetic evaluation of the cocaine-kindling 
process. Society for Neuroscience Abstracts . #1 133. 1990. 

Marley, R.J., J. M. Witkin and S. R. Goldberg. Genetic differences in the development of cocaine- 
kindled seizures. NIDA Monograph: Problems of Drue Dependence . 1990. in press. 

Witkin, J.M., S.R. Goldberg & R.J. Marley. Lack of a genetic correlation between the convulsant and 
epileptogenic effects of cocaine and lidocaine. Abstracts of Comm. Prob. Drug Pep. 52nd Ann. S_ci 
Mtc . 1990. 



92 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00101-05 BGL 

Period Covered: January I, 1990 to December 31, 1990 

Title of Project: Pharmacogenetics: Acute Responses to Drug Administration 

Principal Investigators: 



P.I.: 


G.I. Elmer 


Staff Fellow 


BPGL, NJDA, ARC 


Others: 


S.R. Goldberg 


Chief 


BPGL. NJDA, ARC 




M.C. Ritz 


Senior Staff Fellow 


BPGL, NJDA, ARC 




R.J. Mariey 


Staff Fellow 


BPGL, NJDA. ARC 




C.W. Schindler 


Research Psychologist 


BPGL, NJDA, ARC 



Cooperating Units: None 

Lab/Branch: Behavioral Pharmacology and Genetics Laboratory 

Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, 
Baltimore, MD 21224 

Total Man Years: 2.66 Professional: 1.96 Other: 0.70 

Check Appropriate Boxes: 

Human Subjects Human Tissues X_ Neither 

Minors 

Interviews 

Summary of Work: 

The use of inbred strains to estimate genetic correlations across drug-related phenotypes is useful in 
establishing heritable factors in acute response to drugs and in determining traits with common genetic 
mechanisms. Determination of phenotypes with a common or independent mechanisms is important for 
determining drug mechanisms of action, setting criterion for selective breeding, developing predictive 
models of self-administration, and in the determination of individual genotypes at risk for the acute toxic 
effects. The purpose of these studies is to determine sensitivity of inbred strains for opiate-induced 
analgesia, stimulation, sensitization, respiratory depression, hypothermia and straub tail. The degree of 
covariance across genotype may indicate which genetic components influence susceptibility to opiate- 
induced toxicities and to genetic differences in opiate- reinforced behavior. We are also currently 
assessing the influence of serotonergic receptor subtypes on opiate-induced analgesia, hypothermia and 
locomotion. Recent findings include 1) A strong correlation between opiate receptor concentration, 
analgesic sensitivity and drug-intake, 2) Opioid self-administration behavior does not correlate with the 
stimulant effects of opioids and 3) Demonstration that the amount of opioid that maintains drug- 
reinforced behavior and that which produces significant respiratory depression varies greatly as a 
function of genotype. In collaboration with Dr. Uhl of the Neuroscience Branch we are also evaluating 
behaviorally the acute response to opioids in transgenic mice which are over expressing the endogenous 



93 



opioid enkephalin. To date, the level of expression in brain has been insufficient to produce any 
significant behavioral changes. We have also used pharmacogenetic and pharmacological correlations to 
elucidate the biochemical mechanisms mediating toxic responses to cocaine. Using pharmacogenetic 
methods, we have shown genetic differences in sensitivity to cocaine-induced seizures and lethality. We 
also have data suggesting that a single gene associated with serotonergic 5HT2 receptors appears to 
mediate sensitivity to seizures. We have shown that the binding of cocaine and related compounds to 
serotonin transporters appears to have a primary influence on seizures, while dopamine transporters has 
the greatest influence on lethal responses to cocaine. Drug binding to muscarinic and sigma receptors 
appears to attenuate both toxic responses. We are currently assessing the potential efficacy of several 
pharmacotberapeutic strategies on cocaine-induced toxicity, including sigma related drugs and 
antidepressants. 

Publications 

George. F.R. and Ritz, M.C. Cocaine produces locomotor stimulation in SS but not LS mice: 
Relationship to dopaminergic function. Psvchophannacologv . 101, 18-22, 1990. 

George, F.R., Ritz, M.C. and Meisch, R.A. Ethanol produces similar fixed-ratio rate-depressant effects 
in ALKO AA and ANA rats. Advances in Alcohol and Substance Abuse . 9, 31-42, 1990. 

George, F.R., Porrino. L.J.. Ritz, M.C. and Goldberg, S.R. Inbred rat strain comparisons indicate 
different sites of action for cocaine and amphetamine locomotor stimulant effects. Psvchophannacologv , 
in press, 1990. 

George, F.R. and Ritz, M.C. Common mechanisms of reinforcement from alcohol and other drugs. In: 
Alcohol and Alcoholism . Suppl 1, in press, 1990. 

George, F.R. and Ritz. M.C. Cocaine-induced lethality is associated with interactions between dopamine 
transporters and muscarinic and sigma receptors. Journal of Pharmacology and Experimental 
Therapeutics , in press. 1990. 

Kuhar, M.J., Ritz, M.C. and Boja, J.W. The dopamine hypothesis of the reinforcing properties of 
cocaine. Trends in Neuroscience . in press, 1990. 

Ritz, M.C, Boja, J., George, F.R. and Kuhar. M.J. Cocaine binding sites related to drug self- 
administration. In: Problems of Drug Dependence 1989 . Louis S. Harris, Ed. National Institute on Drug 
Abuse Research Monograph, 95, 239-246. 1990. 

Ritz, M.C, Cone, E J. and Kuhar, M.J. Cocaine inhibition of ligand binding at dopamine, norepinephrine 
and serotonin transporters: A structure-activity study. Life Sciences . 46, 635-645, 1990. 

Ritz, M.C., Boja, J., Carroll, F.I., Zaczak, R. and Kuhar, M.J.[ 3 H] WIN 35,065-2: A ligand for cocaine 
receptors in striatum. Journal of Neurochemistrv . 55, 1556-1562, 1990. 

Ritz, M.C. Biochemical genetic differences in vulnerability todrug effects: Is statistically significant 
always physiologically important and vice versa. Advances in Alcohol and Subs tance Abuse, in press. 
1990. 

Ritz, M.C, Kuhar, M.J. and George, F.R. Combined strategies from alcohol and drug research for 
determining the mechanisms of action of abused substances. In: Alcohol Ab use and Alcholism: Recent 
Advances . Van Thiel, D.H. and Tarter, R.E., Eds., Plenum Press, in press, 1990. 



94 



Ritz, M.C.. George, F.R. and Kuhar, M.J. Molecular mechanisms associated with cocaine effects: 
Possible relationships with effects of ethanol. In: Recent Developments in Alcoholism: Alcohol and 
Cncainp- Clinical and Research Issues . Vol X, Gallant, D.M., in press, 1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures are primarily associated with cocaine binding at 
serotonin transporters. Journal of Pharmacology and Experimental Therapeutics , in press, 1 990. 

Abstracts 

Elmer, G.I. and Goldberg, S.R. Genetic factors in the analgesic, stimulant, respiratory depressant and 
reinforcing properties of opioids. British Association for Psvcbopharmacologv . in press 1990. 

Elmer, G.I., Pieper, J.OT)., Goldberg, S.R. and George, F.R. Genetic variation in opiate receptors and its 
relationship to opiate self-administration. Committee on Problems of Drue Dependence , in press 1990. 

George, F.R. and Ritz, M.C. Cocaine-induced lethality: Mediation by dopamine uptake inhibition and 
direct action at muscarinic receptors. The FASEB Journal . #2775, 4, A745, 1990. 

George, F.R. and Ritz, M.C. Serotonin receptor densities appear to influence acquisition of ethanol- 
reinforced behavior. Alcoholism- Clinical and Experimental Research . 15, in press. 1990. 

Ritz. M.C. and George, F.R. Cocaine-induced seizures: Mediation by specific CNS receptors (presented 
at the 1989 annual ACNP meeting). Neuropsvchopharmacologv. 1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures are initiated by serotonin uptake inhibition, but 
attenuated by direct action at sigma and muscarinic receptors. The FASEB Journal . #2776, 4, A745, 
1990. 

Ritz, M.C. and George, F.R. Cocaine-induced seizures and lethality: Mediation by distinct central 
nervous system receptors. Problems of Drue dependence 1990 . Proceedings of the 52nd Annual 
Scientific Meeting, Committee of Problems of Drug Dependence, Inc. National Institute on Drug Abuse 
Research Monograph, in press, 1990. 

Ritz, M.C. and George, F.R. Cocaine toxicity: Distinct neurotransmitter systems are associated with 
seizures and death. Society for Neuroscience Abstracts . #242.8, 16, 581, 1990. 

Ritz, M.C. and George, F.R. Antidepressant drugs appear to enhance cocaine-induced toxicity (presented 
at the 1990 anual ACNP meeting). Neuropsvcopharmacology . in press, 1990. 



95 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00102-05 BGL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Phaimacogenetic Factors in Drag Reinforced Behavior 

Principal Investigators: 

P.I.: G.I.Elmer Staff Fellow BPGL, NIDA, ARC 

Others: S.R.Goldberg Chief BPGL, NIDA, ARC 

Cooperating Units: None 

Lab/Branch: Behavioral Pharmacology and Genetics Laboratory 

Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drag Abuse, 
Baltimore. MD 21224 

Total Man Years: 1.16 Professional: 0.66 Other: 0.50 

Check Appropriate Boxes: 

Human Subjects Human Tissues X Neither 

Minors 

Interviews 

Summary of Work: 

The objectives of the proposed research are to identify and study factors that control drag reinforced 
behavior using genetically divergent mouse and rat populations. The methodology and principles of 
operant conditioning and pharmacogenetic analysis will be used The studies will be limited to 
conditions in which the drag is taken orally and functions as a positive reinforcer. The focus will be on 
the variables that control drag reinforced behavior, especially genetic variables, but also including 
pharmacological variables and environmental variables, e.g., drag concentration and fixed ratio size. The 
proposed studies are important because ( 1 ) drag intake will be examined under conditions in which it is 
taken orally and functions as a reinforcer, (2) they will explore genetic and environmental factors and 
their interactions which contribute to drag self-administration; and (3) the use of genetically defined 
animals will provide information concerning the degree to which genetic factors regulate drag-seeking 
behavior. These studies complement ongoing investigations into the genetic factors affecting the acute 
response to drags of abuse and will contribute the determination of traits with common or independent 
genetic backgrounds. In addition these studies are important in the development of predictive models of 
self-administration behavior and the determination of individual genotypes at risk for the acute toxic 
effects of the drag. Recent findings included 1) Developing oral cocaine self-administration procedures, 
2) Defining genetic differences in reinforcement from opiates; and 3) Determining common and 
independent genetic factors in the acute, toxic and reinforcing effects of opiates. 



96 



Publications 

Elmer, G.I. and George, F.R. Role of prostaglandin synthetase in the rate depressant effects and narcosis 
caused by ethanoi. Journal ol Pharmacology and Experimental Therapeutics , in press, 1990. 

Elmer, G.I.. Meisch. R.A., Goldberg, S.R. and George, F.R. Ethanoi self- administration in long sleep 
and short sleep mice: Evidence for genetic independence of neurosensitiviry and reinforcement. Journal 
of Pharmacology and Experimental Therapeutics . 254(3), 1054-1062, 1990. 

George, F.R., Ritz. M.R.. and Elmer. G.I. The role of genetics in drug dependence. In: The Biological 
Basis of Drug Tolerance and Dependence . Pratt. J. (ed.), Academic Press, New York, in press. 

George, F.R., Elmer, G.I., Meisch, R.A. and Goldberg, S.R. Orally delivered cocaine functions as a 
positive reinforcer in C57BL/6J mice. Pharmacology Biochemistry and Behavior , in press, 1990. 



97 



2. Psychobiology Laboratory - Jonathan L. Katz, Ph.D., Chief 



Overview 

A major portion of the research efforts of the Psychobiology Laboratory are directed at examining the 
pharmacology of cocaine and the neurotransmitter systems that are involved in the effects of cocaine. In 
addition, there are continuing projects on benzodiazepines and amphetamines. One goal is to develop a 
better understanding of pharmacological mechanisms involved in the behavioral effects of cocaine as 
well as the toxicity that occurs at high doses. Another focus is the development of new pharmacological 
approaches to the treatment of cocaine abuse. Studies of cocaine mechanisms have examined the 
behavioral effects of compounds acting on specific components of the dopamine system. Studies within 
each of these areas have obvious implications for studies within the other areas. 



Summary of Ongoing Research 

A. Basic Mechanisms of cocaine effects 

The primary findings and implications for the current year and future directions are: ( 1) Both Dl and D2 
receptor systems appear to be involved in many of the behavioral effects of cocaine, though neither 
agonist actions at these receptors alone fully reproduces cocaine effects. Non-dopaminergic effects may 
be important components of the pharmacological profile cocaine. (2) The lethal effects of cocaine 
administration appear to principally involve peripheral rather than central nervous system mechanisms. 
The specific pathophysiological processes involved however, remain to be determined. (3) An influence 
of cholinergic systems on the behavioral effects of cocaine has been suggested. Certain muscarinic 
antagonists can mimic some aspects of the behavioral pharmacology of cocaine. Efforts are now being 
directed toward defining the pharmacological nature of the stimulant effects of these drugs. (4) 
Differences in the involvement of postsynaptic dopamine receptors in the effects of various stimulants 
have been established. The differential modulation of these stimulants by GABAergic agents is under 
investigation. (5) Dl receptors appear to be preferentially involved in the lethal effects of cocaine. 
Detailed documentation of the role of dopamine receptor subtypes in cocaine toxicity is a continuing 
experimental effort. (6) Agonist activity at mu-opioid receptors confers stereospecific protection against 
cocaine lethality. (7) Modulation of excitatory amino acid neurotransmission can profoundly alter the 
convulsive effects of cocaine. Ongoing efforts are directed at elucidating the responsible components of 
this system. (8) Tolerance that develops to the behavioral effects of cocaine does not confer cross 
tolerance to all dopamine uptake inhibitors. 

In collaboration with the Neuroendocrinology Laboratory and the Neuroscience Branch studies have been 
conducted to test whether chronic administration of cocaine, or its withdrawal, produces changes in 
neuroendocrine or receptor binding parameters in the rat. To date we have found that repeated injections 
of cocaine causes a hyperprolactinemia. and increasses neurotensin binding in the prefrontal cortex. 
Preliminary results also indicate that binding to the dopamine transporter is decreased in the nucleus 
accumbens after chronic cocaine administration. These results may relate to cocaine dependence, 
tolerance or sensitization. 

B. Drug Development 

These studies are designed to provide preclinical information for the development of medications to be 
used in the treatment of drug abuse. The primary focus of this work is to determine pharmacological 
means for modulating behavioral and toxic actions of abused compounds, and to evaluate new chemical 
entities (synthesized in-house and from outside sources) for safety and efficacy in the design of potential 
rational treatment strategies. 



98 



The primary Undines and implications for the current year and future directions are: (1) Buprenorphine 
protects against the lethal effects of cocaine providing drug-interaction safety information for future 
studies of efficacy in cocaine abuse treatment. (2) Two classes of compounds which block the N-methyl- 
D-aspartate receptor protect against the high-dose convulsant effects of cocaine that are insensitive to 
standard anticonvulsants. These studies suggest a novel avenue for developing new treatments for 
seizures induced by cocaine overdose. (3) A novel class of compounds which are capable of blocking the 
stimulant effects of cocaine has been identified. Ongoing studies will determine whether other effects of 
cocaine related to its abuse and toxicity can be similarly altered. (4) A syndrome of hypersensitivity to 
dopaminergic stimulation after withdrawal from high doses of cocaine has been identified. This 
phenomenon is being further studied to determine if it can serve as a model of cocaine dependence for 
evaluating treatment agents. (5) Compounds proposed by NIDA as potential treatments for cocaine abuse 
have been examined in preclinical screens for safety and efficacy; these studies are in progress. 

C. Behavioral pharmacology of dopamine systems 

Several of our previous findings suggested that dopamine antagonists, acting selectively at either Dl or 
D2 receptors, did not robustly antagonize several behavioral effects of cocaine. In response to these 
findings we have examined the interactions of specific Dl and D2 agonists and antagonists. These studies 
indicate that: ( 1 ) the D2 antagonist, sulpiride, is an effective antagonist of some behavioral effects of the 
D2 agonist, quinpirole in primates; (2) the D2 antagonist, haloperidol was not an effective antagonist of 
the D2 agonist, quinpirole, in rats; (3) the Dl antagonist. SCH 23390, is not effective as an antagonist of 
the Dl agonist SKF 38393 in either rats or primates. We are therefore, pursuing some further studies of 
the interactions of these drugs using several behavioral endpoints. These studies will also pursue a 
number of different compounds as agonists and antagonists of Dl and D2 receptors in order to more fully 
characterize the behavioral pharmacology of these drugs and the contribution of Dl dopamine receptor 
systems to the pharmacology of cocaine. 

We have initiated several studies of the effects of cocaine analogs in collaboration with the Neuroscience 
Branch. One of these studies has examined a drug that binds irreversibly to the cocaine binding site. This 
compound, p_aja-isothiocyanato- benzyolecgonine methyl ester (para-isococaine), has been administered 
into the nucleus accumbens in rats. Following this treatment, cocaine which normally produces a large 
stimulation in locomotor activity, was inactive. In contrast, apomorphine, a direct receptor agonist, 
retained activity. These studies are being followed in order to more fully characterize this effect. 
Obviously, these results may have important consequences in a pharmacological analysis of mechanisms 
of cocaine effects. 

Several antidepressants that block the uptake of dopamine do not appear to be widely abused. In addition, 
several of these compounds have been studied in displacement studies and have not fully displaced 
cocaine from its binding sites. This residual cocaine binding may represent binding to another site that 
may be responsible for the reinforcing effects of cocaine and may differentiate cocaine and non-abused 
dopamine uptake blockers. In order to examine this possibility, the discriminative effects of cocaine and 
other dopamine uptake blockers will be compared. 

Several studies are currently being planned on the neuroanatomical substrates of the psychomotor 
stimulant effects of cocaine. Intracerebral injections will be used to investigate the pharmacological 
nature of neuroanatomical sites related to the actions of cocaine. 

Acetylcholine and GABA are found in high concentrations in the nucleus accumbens. These studies will 
investigate the interconnections of these systems with dopaminergic neurons by injecting cholinergic or 
GABAergic agonists and antatgonists into mesolimbic sites, and testing their effects on locomotor 
activity. This behavior appears to be a good model for psychomotor stimulant action. Important findings 
obtained with locomotor stimulation will be advanced to studies of cocaine self-administration. These 



99 



studies will provide leads for pharmacological modulations of other transmitter systems that may control 
the behavioral effects of cocaine. 



100 



Articles Published or in Press 

Katz, J.L. Effects of drugs on stimulus control of behavior under schedules of reinforcement. In T. 
Thompson. P.B. Dews and J.E. Barrett (Eds.). Advances in Behavioral Pharmacology . Vol. 7. Hillsdale. 
NJ: Lawrence Earlbaum Associates, pp. 13-38., 1990. 

Katz, J.L., Winger, G.D. and Woods. J.H. Abuse liability of benzodiazepines. In I. Hindmarch, G. 
Beaumont. S. Brandon. B.E. Leonard (Eds.). Benzodiazepines: Current concepts . Chichester, U.K.: John 
Wiley and Sons. pp. 181-198, 1990. 

Schindler, C.W., Wu, X.-Z., Su, T.-P., Goldberg, S.R. and Katz, J.L. Enhanced sensitivity to the 
behavioral effects of naltrexone in rats. Journal of Pharmacology and Experimental Therapeutics . 252: 8- 
14. 1990. 

Witkin. J.M., Ricaurte. G.A. and Katz, J.L. Behavioral effects of N-methylamphetamine and N.N- 
dimethylamphetamine in rats and squirrel monkeys. Journal of Pharmacology and Experimental 
Therapeutics . 253: 466-474, 1990. 

Katz. J.L. Models of relative reinforcing efficacy of drugs and their predictive utility. Behavioural 
Pharmacology . 1: 283-301, 1990. 

Katz, J.L., Tirelli, E. and Witkin, J.M. Stereoselective effects of cocaine. Behavioural Pharmacology . 1: 
347-353, 1990. 

Katz, J.L., Dworkin, S.I., Dykstra. L.A., Carter, R.B. and Witkin. J.M. Some behavioral effects of 
repeated d-amphetamine administrations. Drug Development Research . 20: 31-41, 1990. 

Katz, J.L. Benzodiazepine use and abuse. Maryland Pharmacist . 66: 23-28, 1990. 

Witkin, J.M. and Katz, J.L. Analysis of behavioral effects of drugs. Drug Development Research . 20: 
389-409, 1990. 

Katz, J.L. and Witkin, J.M. Effects of cocaine alone and in combination with selective dopamine 
antagonists in the squirrel monkey. Psvchopharmacology . [In Press]. 

Spear, D.J., Muntaner, C, Goldberg, S.R. and Katz, J.L. Methohexital and cocaine self-administration 
under fixed-ratio and second order schedules. Pharmacology Biochemistry and Behavior . [In Press]. 

Katz, J.L. Psychoactive drugs: Tolerance and sensitization. Trends in Neuroscience . 13: 158-159, 1990. 

Katz, J.L. and Goldberg, S.R. Second-order schedules of drug injection: Implications for understanding 
reinforcing effects of abused drugs. In N.K. Mello (Ed). Advances in Substance Abuse. Behavioral and 
Biological Research . Vol. 4. London: Jessica Kingsley Publishers, Ltd., [In Press]. 

Katz, J.L., Winger, G.D. and Woods, J.H. Abuse liability of benzodiazepines and 5-HTia agonists. In R. 
J. Rodgers and S. J. Cooper (Eds.) 5-HTia Agonists. 5-HTj Antagonists and Benzodiazepines: Their 
Comparative Behavioural Pharmacology . Chichester, U.K.: John Wiley and Sons, [In Press]. 

Katz, J.L., Sharpe, L., Jaffe, J.H., Shores, E.I. and Witkin, J.M. Discriminative stimulus effects of inhaled 
cocaine in squirrel monkeys. Psvchopharmacology . [In Press]. 

Katz, J.L. A review of testing and evaluation of drugs of abuse. Behavioural Pharmacology . [In Press]. 



101 



Spear, D.J. and Katz, J.L. Cocaine and food as reinforcers: Effects of reinforcer magnitude and response 
requirement under second-order and second-order progressive- ratio schedules. Journal of the 
Experimental Analysis of Behavior, [In Press]. 

Witkin. J.M. Behavioral pharmacology of compounds affecting muscarinic cholinergic receptors. In J. 
E. Barrett, T. Thompson, and P. B. Dews (Eds.), Advances in Behavioral Pharmacology, vol. 7, 
Hillsdale, NJ. Lawrence Erlbaum, 79-118, 1990. 

Witkin, J.M. and Perez, L.A. Comparison of effects of buspirone and gepirone with benzodiazepines and 
antagonists of dopamine and serotonin receptors on punished behavior of rats. Behavioural 
Pharmacology , 1: 247-254, 1990. 

Genovese, R.F., Elsmore, T.F. and Witkin, J.M. Relationship of behavioral effects of aprophen, atropine 
and scopolamine to antagonism of behavioral effects of physostigmine. Pharmacology Biochemistry and 
Behavior . 37: 117-122, 1990. 

Witkin, J.M. and Goldberg, S.R. Effects of cocaine on locomotor activity and schedule-controlled 
behaviors of inbred rat strains. Pharmacology Biochemistry and Behavior 37: 339-342, 1990. 

Terry, P. and Oakley, DA. The effects of cortical or hippocampal damage on behavioural flexibility in 
the rat. Psvchobiologv , 18: 404-415, 1990. 

Terry, P. and Salmon, P. Biphasic effects of propranolol on a temporal generalization gradient in the rat. 
Journal of Psvchopharmacologv . 4: 63-68. 1990. 

Terry, P., Wray, N. and Salmon P. Acute and chronic effects of propranolol on extinction of rewarded 
running in the rat. Pharmacology. Biochemistry and Behavior . 36: 249-253, 1990. 

Christie, D., Terry, P. and Oakley. D.A. The effects of unilateral anteromedial cortex lesions on prey- 
catching and spatio-motor behaviour in the rat. Behavioural Brain Research . 37: 263-268, 1990. 

Marsland, A.. Stanford, S.C., Terry, P. and Salmon, P. Effects of propranolol on, and noradrenergic 
correlates of, the response to nonreward. Pharmacology, Biochemistry and Behavior . 35: 41-46, 1990. 

Terry, P. and Salmon, P. Anxiolytic-like action of beta-blockers: effects of stimulus salience. 
Pharmacology. Biochemistry and Behavior . [In Press]. 

Terry, P. Review of "Nerve Cells and Animal Behaviour" by D. Young, in Quarterly Journal of 
Experimental Psychology , 42: 218-220, 1990. 

Pilotte, N.S., Sharpe, L.G. and Dax, E.M. Multiple, but not acute, infusions of cocaine alter the release of 
prolactin in male rats. Brain Res. 512: 107-1 12, 1990. 

Sharpe, L.G. and Jaffe, J.H. Ibogaine fails to reduce naloxone-precipitated withdrawal in the morphine- 
dependent rat. NeuroReport 1: 17-19, 1990. 

Sharpe, L.G. Separate neural mechanisms mediate sufentanil-induced pupillary responses in the cat. L 
Pharmacol. Exp. Ther, [In Press]. 

Sze kely, J.L, Sharpe, L.G. and Jaffe, J.H. Dextromethorphan inhibits but dextrorphan potentiates 
behavior induced by PCP and ketamine in rats. NIDA Res. Monogr . [In Press]. 



102 



Abstracts Published or in Press 

Katz, J.L.. Witkin, J.M. and Tirelli, E. Stereoselective effects of cocaine. Paper presented to the Annual 
Meeting of the Behavioral Pharmacology Society, 1990. 

Katz, J.L., Griffiths, J.W. and Witkin, J.M. Lack of cross tolerance to dopamine agonists in cocaine- 
tolerant rats. Psvchopharmacologv . 101: S28, 1990. [Paper presented to the Third Biennial Meeting of 
the European Behavioural Pharmacology Society, 1990.] 

Witkin, J.M. and Katz, J.L. Discriminative stimulus effects of flumazenil in pigeons. 
Psvchopharmacologv . 101: S63. 1990. [Paper presented to the Third Biennial Meeting of the European 
Behavioural Pharmacology Society, 1990.] 

Schindler. C.W., Goldberg, S.R. and Katz, J.L. Pharmacological specificity of enhanced sensitivity to 
naltrexone in rats. Paper presented to the 1990 meeting on the American Psychological Association. [In 
Press]. 

Katz, J.L., Nichols, D.E. and Witkin. J.M. Effects of dopamine receptor-selective drugs in rats trained to 
discriminate cocaine from saline. Abstracts pi the 20th Annual Meeting of the Society for Neuroscience . 
p. 253.. 1990. 

Martello, MB., Martello, A.L., Katz, J.L. and Ricaurte, GA. MDMA-treated monkeys continue to show 
brain serotonergic deficits eighteen months after drug treatment. Abstracts of the 20th Annual Meeting of 
the Society for Neuroscience . 1990. 

Schindler, C.W., Wu, X.-Z., Su, T.-P., Thomdike, E.B., Goldberg, S.R. and Katz, J.L. Enhanced 
sensitivity to naltrexone in rats: Effects on salivation and opioid receptor binding. Abstracts of the 20th 
Annual Meeting of the. Society for Neuroscience , 1990. 

Schindler, C.W., Telia, S.R., Katz, J.L. and Goldberg, S.R. Effects of cocaine and cocaine methiodide on 
cardiovascular function in squirrel monkeys. FASEB Journal . 5: A1587, [In Press]. 

Pell6 n, R. and Katz, J.L. Scratching induced by dopamine D-2 agonists in squirrel monkeys. Submitted 
to the 1991 Meeting of the American Psychological Association. Pharmacology Biochemistry and 
Behavior [In Press). 

Witkin, J. M., Witkin, K. M. and Chiang, P. K. Azaprophen: A novel structural benzilate antimuscarinic 
with unique behavioral activity. European Journal of Pharmacology 1944, 1990. 

Witkin, J. M. and Katz, J. L. Discriminative stimulus effects of flumazenil in pigeons. 
Psvchopharmacologv 101: (Suppl) S63, 1990. 

Katz, J. L., Griffiths, J. W. and Witkin, J. M. Lack of cross tolerance to dopamine agonists in cocaine- 
tolerant rats. Psvchopharmacologv 101: (Suppl) S28, 1990. 

Witkin, J. M. and Tortella, F. C. Blockade of the convulsant effects of cocaine by MK- 801 in a 
diazepam-insensitive mouse model. Society for Neuroscience Abstracts 16: 305, 1990. 

Katz, J. L., Nichols, D. E. and Witkin, J. M. Effects of dopamine receptor-selective drugs in rats trained 
to discriminate cocaine from saline. Society for Neuroscience Abstracts 16: 253, 1990. 



103 



Marley, R. J., Witkin, J. M. and Goldberg, S. R. A pharmacogenetic evaluation of the cocaine kindling 
process. Society for Neuroscience Abstracts 16: 305, 1990. 

Terry, P. A comparison of cocaine-induced activity patterns following single or cumulative dosing 
regimens, Society for Neuroscience. Oct. 1990. 

Sharpe. L. G. and Jaffe, J. H. Ibogaine fails to reduce naloxone-precipitated withdrawal in the morphine- 
dependent rat. Abstract presented at CPDD, June, 1990. 

Pilotte, N. S., Mitchell, W. M, Sharpe, L. G., De Souza, E. B. and Dax. E. M. Chronic cocaine 
administration and withdrawal from cocaine modify central neurotensin receptors in rats. Abstract 
presented at CPDD, June, 1990. 

Jaffe, A. B., Weinhold, L. L. and L. G. Sharpe. Factors influencing self- dministration of aerosol 
sufentanil in rats. Abstract presented at CPDD. June, 1990. 

Szekely, J. I., Sharpe, L. G. and Jaffe, J. H. Dextromethorphan inhibits but dextrophan potentiates 
behavior induced by PCP and ketamine in the rat. Abstract presented at CPDD, June, 1990. 

Sharpe, L. G., Pilotte, N. S., Mitchell, W. M., De Souza, E. B. and Dax, E. M. Withdrawal from chronic 
cocaine decreased dopamine transporter sites in the rat nucleus accumbens (NAc). Abstract presented at 
Soc. Neurosci. Nov. 1990. 

Pilotte, N. S., Sharpe, L. G. and Dax E. M. Chronic cocaine modifies growth hormone release after 5- 
hydroxytryptophan in male rats. Abstract presented at Soc. Neurosci. Nov. 1990. 



104 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

Period Covered: January 1. 1990 to December 31, 1990 
Title of Project: Basic Mechanisms of cocaine effects. 
Principal Investigators: 



Z01 DA 00103-01 PBL 



Co-PI: 
Co-PI: 

Others: 



J.L. Katz 
J.M. Witkin 

L. Sharpe 
P. Terry 
E. Tirelli 



Chief 

Research Psychologist 

Research Psychologist 
Visting Fellow 
Visiting Fellow 



PBL, NTDA, ARC 
PBL, NTDA, ARC 

PBL, NIDA, ARC 
PBL, NDDA, ARC 
PBL, NIDA, ARC 



Cooperating Units: None 

Lab/Branch: Psychobiology Laboratory, Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse 
Baltimore, Maryland 21224 



Total Man Years: 2.9 Professional: 



Check Appropriate Boxes: 



1.9 



Other: 1.0 



Human Subjects 

Minors 

Interviews 



Human Tissues 



X Neither 



Summary of Work: 

Studies of the basic mechanisms of the effects of cocaine have utilized several behavioral procedures. 
The stimulant effects of cocaine are assessed in studies of locomotor activity in rodents, as well as studies 
of learned operant behavior. In addition, the subjective effects of cocaine are examined in studies of the 
discriminative stimulus effects, and reinforcing effects of cocaine. Additional studies are conducted 
assessing cocaine overdose toxicity in which the lethal and convulsive effects of cocaine are assessed. 

Initial studies have investigated the roles of dopamine receptor subtypes in the effects of cocaine. These 
studies have indicated that both Dl and D2 receptor systems appear to be involved in the discriminative 
effects of cocaine, however, agonist actions at only one of these receptors does not appear to be sufficient 
to fully reproduce the subjective effects of cocaine. Results of these studies are forming the basis for 
further studies that will examine the effects of several of these dopaminergic agents on cocaine self- 
administration. Previous studies from this laboratory have indicated that the toxic effects of cocaine may 
be primarily mediated by actions at Dl dopamine receptors. Recent further studies have indicated that 
these toxic effects of cocaine appear to principally involve peripheral rather than central nervous system 
mechanisms. The toxic effects of cocaine may be influenced by actions mediated by several other 
neurotransmitter systems besides the dopamine system. Studies have indicated that agonist actions at mu- 
opioid receptors can protect against the lethal effects of cocaine. This action may be the basis for the 
potential therapeutic effects of buprenorphine as a cocaine abuse treatment. In addition, the convulsive 



105 



effects of cocaine can be attenuated by the administration of excitatory amino acid antagonists, as well as 
modulators of these systems. 

Other studies have investigated the behavioral and physiological effects of repeated administration of 
cocaine. The tolerance that develops to the effects of cocaine on learned operant behavior is relatively 
minor compared to that occurring with other drugs of abuse. It appears that learning plays a significant 
role in the development of tolerance to cocaine. Along with that tolerance there can be physiological 
changes. Repeated injections of cocaine can produce a hyper-prolactinemia, and increases neurotensin 
binding in the prefrontal cortex (where neurotensin is co-localized with dopamine). Preliminary results 
also indicate that binding to the dopamine transporter is decreased in the nucleus accumbens. 

Articles Published or in Press: 

Katz, J.L. Effects of drugs on stimulus control of behavior under schedules of reinforcement. In T. 
Thompson, P.B. Dews and J.E. Barrett (Eds.). Advances in Behavioral Pharmacology . Vol. 7. Hillsdale, 
NJ: Lawrence Earlbaum Associates, pp. 13-38, 1990. 

Katz, J.L. Models of relative reinforcing efficacy of drugs and their predictive utility. Behavioural 
Pharmacology , 1: 283-301. 1990. 

Katz, J.L., Tirelli. E. and Witkin. J.M. Stereoselective effects of cocaine. Behavioural Pharmacology . 1: 
347-353, 1990. 

Katz, J.L. and Witkin, J.M. Effects of cocaine alone and in combination with selective dopamine 
antagonists in the squirrel monkey. Psvchopharmacologv . [In Press]. 

Spear, D.J., Muntaner, C, Goldberg, S.R. and Katz, J.L. Methohexital and cocaine self-administration 
under fixed-ratio and second order schedules. Pharmacology Biochemistry and Behavior . [In Press]. 

Katz. J.L. and Goldberg, S.R. Second-order schedules of drug injection: Implications for understanding 
reinforcing effects of abused drugs. In N.K. Mello (Ed.). Advances in Substance Abuse. Behavioral and 
Biological Research , Vol. 4. London: Jessica Kingsley Publishers, Ltd., [In Press]. 

Katz, J.L., Sharpe, L., Jaffe, J.H., Shores, E.I. and Witkin, J.M. Discriminative stimulus effects of inhaled 
cocaine in squirrel monkeys. Psvchopharmacologv . [In Press]. 

Spear. D.J. and Katz, J.L. Cocaine and food as reinforcers: Effects of reinforcer magnitude and response 
requirement under second-order and second-order progressive-ratio schedules. Journal of the 
Experimental Analysis of Behavior . [In Press]. 

Witkin, J. M. and Goldberg, S. R. Effects of cocaine on locomotor activity and schedule-controlled 
behaviors of inbred rat strains. Pharmacology Biochemistry and Behavior 37: 339-342, 1990. 

Pilotte, N.S., Sharpe, L.G. andDax. E.M. Multiple, but not acute, infusions of cocaine alter the release of 
prolactin in male rats. Brain Res . 512: 107-112, 1990. 

Abstracts Published or in Press: 

Katz, J.L., Witkin, J.M. and Tirelli, E. Stereoselective effects of cocaine. Paper presented to the Annual 
Meeting of the Behavioral Pharmacology Society, 1990. 

Katz, J.L., Griffiths, J.W. and Witkin, J.M. Lack of cross tolerance to dopamine agonists in cocaine- 
tolerant rats. Psvchopharmacologv . 101: S28, 1990. Paper presented to the Third Biennial Meeting of the 
European Behavioural Pharmacology Society, 1990. 



106 



Schindler, C.W., Telia, S.R., Katz, J.L. and Goldberg, S.R. Effects of cocaine and cocaine methiodide on 
cardiovascular function in squirrel monkeys. FASEB Journal . [In Press]. 

Terry, P., Witkin, J.M. and Katz, J.L. Changes in the stimulus effects of cocaine with training dose. 
Submitted to the 1991 Meeting of the American Psychological Association. Pharmacology Biochemistry 
and Behavior [In Press]. 

Katz, J. L., Griffiths, J. W. and Witkin, J. M. Lack of cross tolerance to dopamine agonists in cocaine- 
tolerant rats. Psvchopharmacology 101: (Suppl) S28, 1990. 

Katz, J. L., Nichols, D. E. and Witkin, J. M. Effects of dopamine receptor-selective drugs in rats trained 
to discriminate cocaine from saline. Society for Neuroscience Abstracts 16: 253, 1990. 

Marley. R. J., Witkin. J. M. and Goldberg, S. R. A pharmacogenetic evaluation of the cocaine kindling 
process. Society for Neuroscience Abstracts 16: 305, 1990. 

Pilotte. N. S.. Mitchell, W. M.. Sharpe, L. G, De Souza, E. B. and Dax, E. M. Chronic cocaine 
administration and withdrawal from cocaine modify central neurotensin receptors in rats. Abstract 
presented at CPDD, June, 1990. 

Sharpe, L. G, Pilotte, N. S., Mitchell, W. M. De Souza, E. B. and Dax. E. M. Withdrawal from chronic 
cocaine decreased dopamine transporter sites in the rat nucleus accumbens (NAc). Abstract presented at 
Soc. Neurosci. Nov. 1990. 

Pilotte, N. S., Sharpe, L. G. and Dax E. M. Chronic cocaine modifies growth hormone release after 5- 
hydroxytryptophan in male rats. Abstract presented at Soc. Neurosci. Nov. 1990. 



107 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00104-01 PBL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Drug Development 

Principal Investigators: 

Co-PI: J.M. Wilkin Research Psychologist PBL, NIDA, ARC 

Co-PI: J.L. Katz Chief PBL, NIDA, ARC 

Others: E. Tirelli Visiting Fellow PBL, NIDA, ARC 

Cooperating Units: None 

Lab/Branch: Psychobiology Laboratory, Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse 
Baltimore, Maryland 21224 

Total Man Years: 1.9 Professional: .9 Other: 1.0 

Check Appropriate Boxes: 

Human Subjects Human Tissues X Neither 

Minors 

Interviews 

Summary of Work: 

These studies are designed to provide preclinical information for the development of medications to be 
used in the treatment of drug abuse. This work utilizes leads developed in studies of the basic 
mechanisms of the effects of cocaine in order to further develop pharmacological means for modulating 
behavioral and toxic actions of cocaine. New chemical entities (synthesized in-house and from outside 
sources) for safety and efficacy in the design of potential rational treatment strategies. 

The primary findings and implications for the current year and future directions are: (1) Buprenorphine 
protects against the lethal effects of cocaine. The mechanism by which this effect occurs appears to be 
mu-receptor mediated. The effect was stereospecific, and did not occur in a strain of mice that was mu- 
receptor deficient. Other mu agonists were equally efficacious with potencies that varied according to 
their relative affinities for the mu receptor. 

Drugs that competitively or non-competitively block the N-methyl-D-aspanate receptor were found to 
protect against the high-dose convulsant effects of cocaine. This protection was afforded at doses of 
cocaine that produced effects that could not be antagonized by standard anticonvulsants. These studies 
suggest a novel avenue for developing new treatments for seizures induced by cocaine overdose. Work on 
other modulators of this receptor and its associated mechanisms are in progress. 

Other studies are being conducted in order to develop a model for assessing cocaine withdrawal 



108 



phenomena. A syndrome of hypersensitivity to dopaminergic stimulation after withdrawal from high 
doses of cocaine has been characterized. If this phenomenon proves to be a valid model of cocaine 
dependence it will be utilized for evaluating potential cocaine-withdrawal treatment agents. 

Articles Published or in Press: 

Witkin, J.M. and Katz, J.L. Analysis of behavioral effects of drugs. Drug Development Research . 20: 
389-409, 1990. 

Katz, J.L. A review of testing and evaluation of drugs of abuse. Behavioural Pharmacology . [In Press]. 

Sze kely, J.L, Sharpe, L.G. and Jaffe, J.H. Dextromethorphan inhibits but dextrorphan potentiates 
behavior induced by PCP and ketamine in rats. NIDA Res. Monogr . [In Press]. 

Abstracts Published or in Press: 

Katz. J.L., Nichols. D.E. and Witkin, J.M. Effects of dopamine receptor-selective drugs in rats trained to 
discriminate cocaine from saline. Abstracts of the 20th Annual Meetinp pi the Society for Neuroscience , 
p. 253, 1990. 

Witkin. J. M., Witkin, K. M. and Chiang, P. K. Azaprophen: A novel structural benzilate antimuscarinic 
with unique behavioral activity. European Journal of Pharmacology . 1944, 1990. 

Witkin, J. M. and Tortella, F. C. Blockade of the convulsant effects of cocaine by MK-801 in a 
diazepam -insensitive mouse model. Society for Neuroscience Abstracts . 16: 305, 1990. 

Terry, P. A comparison of cocaine-induced activity patterns following single or cumulative dosing 
regimens. Society for Neuroscience . Oct. 1990. 

Szekely, J. I., Sharpe, L. G. and Jaffe, J. H. Dextromethorphan inhibits but dextrophan potentiates 
behavior induced by PCP and ketamine in the rat. Abstract presented at CPDD, June. 1990. 



109 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: Behavioral pharmacology of dopamine systems 

Principal Investigators: 

Co-PI: J.L. Katz Chief PBL, NTDA, ARC 

Co-PI: J.M. Witkin Research Psychologist PBL, NIDA, ARC 

Others: L. Sharpe Research Psychologist PBL, NIDA, ARC 

P. Terry Visting Fellow PBL, NIDA, ARC 

E. Tirelli Visting Fellow PBL, NIDA. ARC 

Cooperating Units: None 

Lab/Branch: Psychobiology Laboratory, Preclinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse 
Baltimore, Maryland 21224 



Z01 DA 00105-01 PBL 



Total Man Years: 2.9 Professional: 1.9 

Check Appropriate Boxes: 

Human Subjects _ 

Minors 
Interviews 



Other: 1.0 



Human Tissues 



X Neither 



Summary of Work: 

These studies are directed at a better understanding of the behavioral effects of drugs acting on dopamine 
neurotransmission. Several previous studies had results indicating that selective Dl or D2 dopamine 
agonists and antagonists were not functioning as they would be expected based on their in vitro 
pharmacology. Subsequent studies have been directed at quantitative assessments of behavioral effects of 
agonist-antagonist interactions. These studies have utilized several different behavioral endpoints, 
including grossly observable behavior, locomotor activity, consummatory behavior, learned operant 
behavior, and drug discrimination. In addition, a series of studies is examining the behavioral effects of 
drugs that are thought to bind irreversibly to the dopamine transporter. These studies will provide answers 
to several basic questions regarding the functioning of this site as well as provide information that may be 
of use in the development of drugs to treat cocaine abuse. Finally, studies are being conducted that 
address the interactions of the dopamine system with cholinergic and GABAergic systems. 



110 



Articles Published or in Press: 

Witkin, J.M., Ricaurte. G.A. and Katz. J.L. Behavioral effects of N-methylamphetamine and N.N- 
dimethylamphetamine in rats and squirrel monkeys. Journal of Pharmacology and Experimental 
Therapeutics . 253: 466-474, 1990. 

Katz, J.L., Dworkin, S.I., Dykstra, L.A., Carter, R.B. and Witkin, J.M. Some behavioral effects of 
repeated d-amphetamine administrations. Drug Development Research . 20: 31-41, 1990. 

Katz, J.L. Psychoactive drugs: Tolerance and sensitization. Trends in Neuroscience . 13: 158-159, 1990. 

Witkin, J. M. Behavioral pharmacology of compounds affecting muscarinic cholinergic receptors. In J. 
E. Barrett, T. Thompson, and P. B. Dews (Eds.), Advances in Behavioral Pharmacology , vol. 7, 
Hillsdale, NJ. Lawrence Erlbaum. 79-118, 1990. 

Witkin, J. M. and Perez, L. A. Comparison of effects of buspirone and gepirone with benzodiazepines 
and antagonists of dopamine and serotonin receptors on punished behavior of rats, Behavioural 
Pharmacology . 1: 247-254, 1990. 

Genovese, R. F., Elsmore, T. F. and Witkin. J. M. Relationship of behavioral effects of aprophen. 
atropine and scopolamine to antagonism of behavioral effects of physostigmine. Pharmacology 
Biochemistry and Behavior . 37: 1 17-122. 1990. 

Abstracts Published or in Press: 

Pell6 n, R. and Katz, J.L. Scratching induced by dopamine D-2 agonists in squirrel monkeys. Submitted 
to the 1991 Meeting of the American Psychological Association. Pharmacology Biochemistry and 
Behavior [In Press). 



Ill 



Clinical Pharmacology Branch 

Jack E. Henningfield, Ph.D., Chief 

Introduction 

The Clinical Pharmacology Branch conducts studies of the effects of drugs in human volunteers to 
provide the basic information necessary to understand, treat, and prevent drug abuse and dependence. 
The Branch is comprised of three laboratories: Biology of Dependence and Abuse Potential Assessment, 
Jack E. Henningfield, Ph.D., Chief; Chemistry and Drug Metabolism, Edward J. Cone, Ph.D., Chief; and 
Neuroendocrinology/ Immunology Laboratory, Jack E. Henningfield. Ph.D., Acting Chief. 

The Clinical Pharmacology Branch emerged from the initial program of basic human research which 
began in the 1930's at the Addiction Research Center in Lexington. Kentucky. It was evident then that 
the reconciliation of clinical reports with scientific theories of drug addiction could best be accomplished 
through the careful observation of human volunteers in a controlled research setting. Drugs were 
administered and withdrawn, and the consequences documented, while minimizing risks to the 
volunteers. Factors likely to be relevant to the course and/or treatment of drug dependence were also 
evaluated, e.g., medications. This program of research constituted the clinical pharmacology program of 
the ARC and today is represented by the Clinical Pharmacology Branch along with the Etiology and 
Treatment Branches. 

The Clinical Pharmacology Branch is well suited to investigate the biological basis of drug abuse and 
dependence by conducting studies that involve observations during the administration and withdrawal of 
pharmacologic agents in human volunteers. There is no administrative obstacle to the laboratories 
working collaboratively to investigate phenomena ranging from metabolism of an administered drug, to 
the modulation of its effects by the neuroendocrine system, to effects of drug deprivation on performance 
and subjective response. The diverse range of expertise and technological support required by such a 
program is available in three laboratories of the branch. 

Much of the present research effort of the Branch represents collaborations among its laboratories as well 
as with other Branches of the ARC. Such multidisciplinary collaborations represent one of the unique 
strengths of scientific exploration at the ARC. The Clinical Pharmacology Branch has also continued in 
its historically important mission of providing a training ground for the development of researchers and 
clinicians. This training mission is administratively stimulated by our efforts to maintain a balance of 
tenured to non-tenured scientists in which approximately one third of the scientists leave the branch every 
two to three years. 

Senior scientists of the Branch are regularly invited to provide lectures and training courses, review 
grants, and provide consultative advice on scientific issues to other Federal agencies and to both national 
and international organizations. Dr. Jack E. Henningfield serves as an advisor to the US Surgeon 
General, Centers for Disease Control, Food and Drug Administration and other agencies on issues 
pertaining to nicotine dependence. Dr. Henningfield collaborates with national and international 
organizations on issues pertaining to the assessment of the abuse liability of chemicals and development 
of drugs. Dr. Edward J. Cone has received national and international recognition for his work on the 
detection of drugs of abuse in biological fluids. His advice is widely sought by federal agencies such as 
the Department of Defense and private industry. He was invited to present his findings on the detection 
of drugs in hair to the second International Congress of Therapeutic Drug Monitoring and Toxicology in 
Barcelona, Spain and to numerous scientific groups in the United States. Dr. Cone also serves as a 
consultant to the National Laboratory Certification Program. Dr. Stephen J. Heishman serves as a 
lecturer and consultant to the Federal Aviation Administration in their drug testing program. Dr. Wallace 
B. Pickworth is a widely recognized author and consultant on the development and evaluation of 
pupilometric devices for possible use in onsite assessment of drug exposure. 



112 



1. Biology of Dependence and Abuse Potential Assessment Laboratory 
- Jack E. Henningfield, Ph.D., Chief 

The Biology of Dependence and Abuse Potential Assessment Laboratory (BDL) assesses the biological 
basis of drug dependence using quantitative experimental procedures of the behavioral and 
pharmacological disciplines; and assesses the abuse liability and physical dependence potential of 
selected compounds. These aims are intended to serve the overall mission of the ARC in providing a 
better foundation for understanding drug dependence and for developing rational approaches for 
preventing and treating drug dependence. 

The BDL evoked out of a tradition of research whose goal was to characterize drug-induced changes in 
behavior and physiologic function; specifically, phenomena such as drug seeking, tolerance, and physical 
dependence. The understanding of these phenomena and their interrelations provides much of the 
pharmacologic and behavioral basis for evolving theories of drug dependence. A practical product of this 
research was the development of standardized procedures to assess the potential of drugs to produce 
dependence, i.e., abuse liability and physical dependence potential tests. Early research by 
Himmelsbach. Frazier, Isbell, Martin, and others, produced fundamental observations upon which much 
of current theory about the understanding and treatment of drug dependence is based. Specific areas of 
exploration included the following: 

a. the relationship between drug administration and development of tolerance, physical dependence 
and changes in mood and behavior, 

b. the use of drug substitution and antagonist administration procedures to study the biologic basis of 
drug dependence and to treat addicted people; 

c. the phenomena whereby drug administration could lead to the alleviation of dysphoric mood states 
or the production of euphoric mood states by the presentation of certain drugs; and 

& patterns of drug seeking in the presence and absence of pharmacologic pretreatment. 

In the course of conduct of these and other basis studies, new strategies of assessment emerged. The 
methods included the use of observer ratings, pupilometry and cardiovascular assessment, and 
electroencephalogram (EEG) to provide objective markers of drug administration, as well as the 
development of new instruments for assessing the effects of drugs on mood, feeling, and behavior. Data 
obtained using such methods and instruments proved not only to be useful in exploration of the basic 
phenomena underlying drug dependence, but also led to objective methods of abuse liability assessment. 
The ability to both quantitatively and qualitatively characterize the clinical pharmacology of substances 
was also fundamental in the development of more selective, safer, and more efficacious agents for the 
alleviation of human disease and suffering. 

Most studies of the BDL are multidisciplinary in nature and involve collabortions with one or more other 
laboratories of the ARC. With such multidisciplinary efforts it is possible to quantitate the subjective 
physiologic, behavioral, electrophysiologic, cognitive, pharmacodynamic, pharmacokinetic, reinforcing, 
aversive, and other effects of drugs, as well as to assess the biologic generality of phenomena by 
comparative animal-human research. 



113 



Publications 

Boren, J. J.. Stitzer, ML. and Henningfield. J.E. Preference among research cigarettes with varying 
nicotine yields. Pharmacol Biochem Behav . 36:191-193. 1990. 

Cohen, C. and Henningfield. J.E. Nicotine dependence: A preventable risk factor for other diseases. 
Journal of General Internal Medicine . 5(Suppl), S73-S78, 1990. 

Cone, E.J. On-site Drug Testing Expediency Versus Accuracy. Employment Testing, BWR :621-28. 
1990. 

Cone. E.J. Testing Human Hair for Drugs of Abuse. I. Individual dose and the profiles of morphine and 
codeine in plasma, saliva, urine and beard compared to drug-induced effects on pupils and behavior. J 
Anal Toxicol . 14:1-7. 1990. 

Cone, E.J., Welch, P, Mitchell, J.M. and Paul, B.D. Forensic Drug Testing for Opiates: I. Detection of 6- 
Acetylmorphine in Urine as an Indicator of Recent Heroin Exposure: Drug and Assay Considerations and 
Detection Times. J Anal Toxicol , 15:1-7, 1990. 

Cone. E.J., Yousefnbejad, D. and Dickerson, S.L. Validity Testing of Commercial Urine Cocaine 
Metaboolite Assays. IV. Evaluation of the EMIR D.A.U.T"* Cocaine Metabolite Assay in A 
Quantitative Mode for Detection of Cocaine Metabolite. J Forensic Sci . 3:786-791, 1990. 

Dax. E.M., Partilla, J.S., Pineyro, M.A. and gregerman, R.I. Altered glucagon- and catecholamine 
hormone- sensitive adeylyl cyclase responsiveness in rat liver membranes induced by manipulation of 
dietary fatty acid intake. Endocrinology 127 (5):2236-2240, 1990. 

Evans, S.M. and Johanson, C.E. Three-choice discrimination among +-amphetamine, fenfluramine and 
saline in pigeons. J Pharmacol Exp Ther . 255:1246-1255. 1990. 

Evans, S.M., Funderburk, F. and Griffiths, R.R. Zolpidem and triazolam in humans: Behavioral effects 
and abuse liability. J Pharmacol Exp Ther . 255:1246-1255. 1990. 

Griffiths, R.R., Evans. S.M., Heishman, S.J., Preston, K.L., Sannerud, C.A., Wolf, B. and Woodson, P.P. 
Low-dose caffeine discrimination in humans. J Pharmacol Exp Ther 252:970-987, 1990. 

Griffiths. R.R.. Evans. S.M., Heishman, S.J., Preston, K.P., Sannerud. C.A., Wolf, B. and Woodson, P.P. 
Low-Dose Caffeine Physical Dependence. J Pharmacol Exp Ther . 255:1123-1132, 1990. 

Haertzen, C.A.. Hickey, J.E., Rose, M.R. and Jaffe, J.H. The relationship between a diagnosis of 
antisocial personality and hostility: Development of an antisocial hostility scale. J Clin P yvchol . 46:679- 
686, 1990. 

Heishman, S.J., Huestis, M.A., Henningfield. J.E. & Cone, E. J. Acute and residual effects of marijuana: 
Profiles of plasma THC levels, physiological, subjective and performance measures. Pharmacol Biochem 
Behav . 37:561-565, 1990. 

Heishman, S.J., Stitzer, ML., Bigelow, G.E. & Liebson, I.A. Acute opioid physical dependence in 
humans: Effect of naloxone at 6 and 24 hours postmorphine. Pharmacol Biochem Behav . 36:393-399, 
1990. 

Henningfield, J.E., Cohen, C, Giovino, G.A. Can genetic constitution affect the "objective" diagnosis of 
nicotine dependence? Am J Public Health 80:1040-1041. 1990. 



114 



Henningfield, J.E., Cohen, C, Pickworth, W.B. Psychopharmacology of nicotine. In: J.D. Slade and C.T. 
Orleans (eds.) Nicotine Addiction: Principles and Management . Oxford University Press. 

Henningfield, J.E., London, E.D. and Benowitz, N.L. Arterial- venous differences in plasma 
concentrations of nicotine after cigarette smoking. JAMA . 263:2049-2050, 1990. 

Henningfield, Ph.D., Radzius, A., Cooper, T.M. and Clayton, R.R. Drinking coffee and carbonated 
beverages blocks absorption of nicotine from nicotine polacrilex gum. JAMA , 264:1550-1564, 1990. 

Huestis, M.A., Heishman, S.J. and Cone, E.J. Profile of Repeated Marijuana Exposure: Plasma and 
Saliva THC Levels, Performance, Physiological and Subjective Effects Following smoking in a 
Controlled Clinical setting, proceedings of the International Association of forensic Toxicologists, Perth, 
Australia, 1990 (submitted). 

Kirby, K.C.. Stitzer, M.L. & Heishman, S.J. Acute opioid physical dependence in humans: Effect of 
varying the morphine-naloxone interval. II. J Pharmacol Exp Ther . 255:730-737, 1990. 

Kumor, KM., Clark, W.C, Janal. M.N. and Haertzen, C.A. Multidimensional scaling of subjective 
judgements of drug similarities among ketocyclazocine, morphine, cyclazocine, naloxone and placebo. 
Pharmacol Biochem Behav , 35:397-404, 1990. 

Lange, W.R., Cone, E.J. and Snyder. F.R. The Association of Hepatitis Delta and Hepatitis B Virus in 
Parental Drug Abusers. Arch Intern Med . 150:365-68, 1990. 

Lathers, CM., Pickworth, W.B., Keefe, D., Spino, M., Agarwal, I. and Tyau, L.S.Y. Cocaine Seizures, 
Arrhythmias and Sudden Death. In: Sudden Death in Epileptic Persons: Occurrence and Possible Causes: 
Autonomic Dysfunction. Cardiac Arrhythmias and Epileptogenic Activity . CM. Lathers and PL. 
Schraeder (eds). Marcel Decker, New York, pp. 417-446, 1990. 

Muntaner. C, Walter, D., Nagoshi. C, Fishbein. D., Haertzen, C.A. and jaffe, J.H. Self-report vs 
laboratory measures of aggression as predictors of substance abuse. Drug and Alcohol Dependence . 
25:1-11, 1990. 

Pickworth, W.B., Gerard-Ciminera, J. and Lathers, CM. Stress, Arrhythmias and Seizures. In: Sudden 
Death in Epileptic Persons: Occurrence and Possible Causes: Autonomic Dysfunction. Cardiac 
Arrhythmias, and Epileptogenic Activity . CM. lathers and P.L. Schraeder (eds). Marcel Decker, New 
York, pp. 393-415. 1990. 

Pickworth, W.B., Herning, R.I., Koeppl, B. and Henningfield, J.E. Dose-dependent atropine-induced 
changes in spontaneous electroencephalogram in human volunteers. Military Medicine 155:166-170, 
1990. 

Pickworth. W.B., Lee, H. and Fudala. P.J. Buprenorphine-induced pupillary effects in human volunteers. 
Life Sciences, 47:1269-1277, 1990. 

Pickworth, W.B., Brown, B.S., Hickey, J.E. and Muntaner, C Effects of self-reported drug use and 
antisocial behavior on evoked potentials in adolescents. Drug and Alcohol Dependence . 25:105-110, 
1990. 

Pilotte, N.S.. Sharpe, L.G., Dax, E.M. Multiple, but not acute, infusions of cocaine alter the release of 
prolactin in male rats. Brain Research . 512:107-112, 1990. 



115 



Ulrichsen, J., Partilla, J.S. and Dax, E.M. Long-term administration of m-chlorophenyl-piperazine 
(mCPP) to rats induces changes in serotonin receptor binding, dopamine levels and locomotor activity 
without altering prolactin and corticosterone secretion. Psvchopharmacology . Submitted. 

Vaupel, D.B., Cone, E.J., Johnson, RE. and Su, T-S. Kappa Opioid Prrtial Agonist Activity of the 
Enkephalin-Like Pentapeptide BW942C Based on Urination and In Vitro Studies in Humans and 
Animals. J Pharmacol Exp Ther . 252:225-34, 1990. 

Weddington, WW., Brown, B.S., Haertzen, C.A., Cone, E.J., Dax, E.M., Herning, R.I. and Michaelson, 
B.S. Changes in Mood, Craving, and Sleep During Acute Abstinence Reported by Male Cocaine 
Addicts: A Controlled Residential Study. Arch Gen Psychiatry . 47:861-868. 1990. 

Abstracts 

Henningfield. J.E. Tobacco as an addicting drug. Statehouse Day for a Tobacco-free Ohio. Columbus. 
Ohio. February 20. 1990. 

Henningfield, J.E. Overview of the Addiction Research Center and scientific needs for increased 
minority involvement in research programs. Fourth Annual Coppin State College Substance Abuse 
Conference. Baltimore, April 27, 1990. 

Henningfield, J.E. Tobacco dependence as an addiction. Presented at the 192nd Annual Meeting of the 
Medical and Chirurgical Faculty of Maryland. Ellicott City, Maryland, May 10, 1990. 

Henningfield. J.E. Pharmacology of nicotine addiction in adolescents. Presented as part of a symposium 
entiUed "Nicotine addiction in adolescents", at the Annual Meeting of the American Psychiatric 
Association, New York City, May 16, 1990. 

Henningfield, J.E. Tobacco addiction. Tobacco in the 90's. Palo Alto, CA, July 25, 1990. 

Henningfield, J.E. Prelude to addiction. Presented in session entided "Preventing minor's access to 
tobacco". STAT (Stop Teenage Addiction to Tobacco) conference, Boston, August 19, 1990. 

Henningfield, J.E. Food and tobacco self-administration: Common theoretical issues and research 
problems. Smoking and Body Weight Conference Workshop, Memphis State University and National 
Heart, Lung, and Blood Institute, Memphis, TN, September 10-13, 1990. 

Henningfield, J.E. Drug self-administration methods in abuse liability evaluation. Barcelona Conference 
on Clinical Abuse Liability Testing. Barcelona, Spain, November 5, 1990. 

Henningfield, J.E. Pharmacologic basis and treatment of nicotine dependence. AMERSA (Association 
for Medical Education and Research in Substance Abuse). Rockville, MD, November 15, 1990. 

Henningfield, J.E. The basis of nicotine dependency in cigarette smokers. Patient-Centered Smoking 
Cessation: An Office Based Model. Sponsored by the Heart of Harlem Cardiovascular Disease Program, 
Harlem Hospital, New York, November 18, 1990. 

Henningfield, J.E. Nicotine: The quintessential mood and performance regulator, and Things you never 
knew could screw up nicotine polacrilex administration. New Directions: The Role of Nicotine Reduction 
Therapy in the 90's. Palm Springs, CA, December 14-15, 1990. 

Cohen, C, Le Houezec, J., Martin, C, Molimard, R. The role of nicotine titration in smoking behavior. 
NIDA Research Monograph Series, Proceedings of the 52nd Annual Scientific Meeting (in press). 



116 



Cone, E.J. and Mitchell, J. Can the NIDA cutoffs for opiate uring screening and confirmation be 
lowered? AAFS, Norfolk, VA. August, 1990. 

Cone, E.J., Darwin, W. and Dickerson, S. Detection of buprenorphine in human biofluids utilizing 
diagnostic product corporations (DPC): Double antibody radioimmunoassay. TIAFT, October 19-23, 
1990. Perth, Western Australia. 

Dax, E.M., Pilotte. N.S. Growth hormone release is altered in men who abruptly cease long-term 
cocaine. Presented at the 2nd International Congress for Neuroendocrinology, June, 1990, Bordeaux, 
France. 

Driscoll. P., Cohen, C, Fackelman, P., Battig, K. Differential ethanol consumption in Roman high- and 
low-avoidance (RHA and RLA) rats, body weight, food intake, and the influence of pre- and postnatal 
exposure to nicotine and/or injection stress. Experientia, 46 A56, 1990. 

Driscoll, P., Cohen, C, Fackelman, P., Lipp, H.B., Battig, K.: Effects of pre- and postnatal injections of 
"smoking doses" of nicotine or vehicle alone, on the maternal behavior and second-generation adult 
behavior of Roman high- and low-avoidance rats. Advances in Pharmacological Sciences (in press). 

Driscoll. P.. Cohen, C, Fackelman, P., Battig, K.: First and second generation effects of pre- and 
postnatal injections of physiological saline (stressor alone), or "smoking doses" of nicotine on the 
maternal behavior of Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats. Behav Genet 20, 
1990 (in press). 

Driscoll, P., Gentsch, C, Fackelman, P., Cohen, C, Battig, K.: Sensitivity to acute ethanol in Roman 
high- and low-avoidance (RHA and RLA) rats: effects of sex, age, and pre- postnatal exposure to ethanol. 
Experientia. 46 A56, 1990. 

Evans, S.M. and Griffiths, R.R. Zolpidem and triazolam in humans: Behavioral effects and abuse 
liability. In: L.S. Harris (Ed.) Problems of Drug Dependence, 1989. NIDA Monograph 95, U.S. 
Government Printing Office, Washington, D.C., 1990, pp. 42-43. 

Fudala. P.J., Johnson, RE., Heishman, S.J., cone, E.J. & Henningfield, J.E. A dose run-up and safety 
evaluation oof nalmefene HC1 in human volunteers. (Abstract: Problems of Drug Dependence 1989, 
NIDA Research Monograph 95, 451-452, 1990). The Committee on Problems of Drug Dependence, Inc.; 
Keystone, Colorado; 19 June 1989. 

Heishman, S.J., Lamb, R.J. & Henningfield, J.E. Discriminative stimulus effects of drugs in humans: 
Stimulants and sedatives. (Abstract: Pharmacology Biochemistry and Behavior, 36, 428, 1990). In 
symposium entiUed, The Current Status of Human Drug discrimination Research. American 
Psychological Association; Boston, MA; August 11, 1990. 

Heishman, S.J. & Stitzer, ML. Time course of repeated naloxone challenge after single morphine doses 
in humans. (Abstract: Problems of Drug Dependence 1989, NIDA Research Monograph 95, 385-386, 
1990). The Committee on Problems of Drug Dependence, Inc.; Keystone, Colorado; 21 June 
1989. 

Heishman, S.J., Snyder, F.R. & Henningfield, J.E. Effect of repeated nicotine administration in 
nonsmokers. (Abstract: Problems of Drug Dependence 1990, NIDA Research Monograph, in press). 
Committee on Problems of Drug Dependence, Inc., Richmond, Virginia; June 11, 1990. 



117 



Heming, R.I.. Brigham, J., Stitzer, M.L.. Glover, B.J., Pickworth, W.B. and Henningfield, J.E. The 
effects of nicotine on information processing: Medicating a deficit. Society for psychophysiological 
Research, Boston: October 1990. (Abstract) 

Huestis, M.A., Heishman, S.J. and Cone, E.J. Plasma and saliva THC levels and behavioral effects 
following repeated marijuana exposure. Society of Forensic Toxicology, Sept. 11-15, 1990. 

Huestis, MA. and Cone, E.J. Time course of detection of salivary 9-THC levels during controlled 
clinical studies of marijuana smoking. The 2nd International Congress of Therapeutic Drug Monitoring 
and Toxicology, Barcelona, Spain, Oct. 9-12, 1990. 

Huestis. M.A.. Heishman, S.J. and Cone, E.J. Profile of repeated marijuana exposure: Plasma and saliva 
THC levels, performance, physiological and subjective effects following smoking in a controlled clinical 
setting. The International Association of Forensic Toxicologists 27th International Meeting, Perth, 
Western Australia, Oct. 19-23, 1990. 

Kirby, K.C., Stitzer, M.L. & Heishman, S. Acute opioid physical dependence in humans: Maximum 
morphine-naloxone interval. (Abstract: Problems of Drug Dependence 1989, N1DA Research 
Monograph 95, 393-394, 1990). The Committee on Problems of Drug Dependence, Inc.; Keystone. 
Colorado: 21 June 1989. 

Mitchell. J.. Paul, B., Welch. P. and Cone, E.J. Clinical studies indicate that morphine is not metabolized 
to codeine in human subjects. AAFS, Cincinnati, OH, Feb. 19-24, 1990. 

Pickworth, W.B., Klein, S.A., Bunker, E.B. and Henningfield, J.E. Assessment of mazindol for abuse 
liability. Committee on Problems of Drug Dependence, NIDA Research Monograph 1990 (in press). 

Pickworth. W.B.. Radzius. A, Welch, P. Opiate-induced changes in dynamic pupillary responses in 
humans. 18th Annual Pupil Colloquim, Berkeley, CA, August 1989. 

Pilotte, N.S., Mitchell, W.M., Sharpe, L.G., De Souza, E.B., Dax, E.M. Chronic cocaine administration 
and withdrawal from cocaine modify central neurotensin receptors in rats. Presented at the 52nd Annual 
Meeting - CPDD. June, 1990, Richmond, VA. 

Pilotte, N.S., Johnson, R.L., Dax, E.M. Chronic coccaine in vivo modifies prolactin release after 
dopamine in vitro. Presented at the 2nd International Congress for Neuroendocrinology, June, 1990, 
Bordeaux, France. 

Pilotte, N.S., Sharpe, L.G., Dax, E.M. Chronic cocaine modifies growth hormone release after 5- 
hydroxytrytophan in male rats. Presented at 20th Ann. Meeting Soc. for Neuroscience, October, 1990, St. 
Louis, MO. 

Sharpe, L.G., Pilotte, N.S., Mitchell, W.M., De Souza, E.B., Dax, E.M. Withdrawal from chronic cocaine 
decreased dopamine transporter sites in the rat nucleus accumbens. Presented at the 20th Annual 
Meeting of Soc. for neuroscience, October, 1990, St. Louis, MO. 

Weddington, W.W., Brown, B.S., Haertzen, C.A., Cone, E.J., Dax, E.M., Heming, R.I., Michaelson, 
B.S. Changes in mood, craving and sleep during acute abstinence by male cocaine addicts. Presented as 
oral communication on June 12, 1990 at meeting of Committee on Problems of Drug Dependence, June 
12, 1990, Richmond, Virginia. 

Weddington. WW.. Haertzen, C.A., et al. Acute abstinence syndrome in male cocaine addicts. 
Presented at the American Psychiatric Association Annual Meeting held in NY, NY on May 12-17, 1990. 



118 



Weddington, WW.. Haertzen, C.A., et al. Reactions by cocaine addicts to HIV-serostatus. Presented at 
the American Psychiatric Association Annual Meeting held in NY. NY on May 12-17, 1990. 



119 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00037-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Physiologic, cognitive and subject effects of commonly abused drugs 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: W.B. Pickworth Pharmacologist, BDL, ARCNIDA 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

X_ Human Subjects Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects with histories of polydrug abuse are studied on the Residential Research Unit to assess the 
effects of several classes of abused drugs on cognitive, subjective and physiologic measures. The main 
purpose of the study is to parametrically compare the sensitivity of various testing instruments across 
several classes of drugs, doses and time. The results are theoretical important because they will evaluate 
the sensitivities of methods used in the drug abuse field The study is of practical importance because it 
evaluates the utility of dynamic pupillography as a drug detection screet. The subject testing has been 
completed (n=9). Preliminary results suggest that pupilometry may identify recent ingestion of opiates, 
marijuana and pentobarbital. The results are submitted for presentation at the 1991 CPDD meeting. 



120 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00028-02 BDL 

January 1. 1990 to December 31, 1990 

Title of Project: Assessment of Mazindol for Abuse Liability 

Principal Investigators: Cooperating Units 

P.I.: J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: W.B. Pickworth Pharmacologist, BDL, ARC.NJDA 

M.J. Kuhar Chief, Neuroscience Branch 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

^X Human Subjects _ Human Tissues Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects with histories of stimulant abuse are studied on the Residential Research Unit to determine the 
abuse liability of mazindol (an anorectant with some psychomotor stimulant properties) to 
methylphenidate (a prototypic psychomotor stimulant with a known potential for abuse). This study was 
performed because mazindol has been used in binding studies aimed at isolating the cocaine receptor. 
The results of these studies indicated that mazindol has high affinity binding at cocaine-sensitive 
dopamine receptor sites. Mazindol is a theoretically interesting drug since its apparent mechanism of 
action (blocks reuptake of norepinephine and dopamine) suggests that it would have abuse potential. 
However, one previous study and limited clinical experience, suggest that it is seldom abused. Therefore 
additional characterization of the clinical pharmacology of mazindol could be of importance in analytic 
efforts as well as drug development. This study is conducted in collaboration with the Neuroscience 
Branch. Subject testing has been completed. Mazindol and methylphenidate increased heart rate and 
diastolic blood pressure and decreased hunger. Mazindol decreased vigor and increased measures of 
fatigue and tired and elevated scores on the PCAG and LSD scales of the ARCI. Methylphenidate did 
not cause the sedative-like effects seen after mazindol. Subjects reported disliking for each drug. These 
data indicate that at doses three times the therapeutic level mazindol poses little abuse potential. On the 
other hand its dysphoric effects call to question the acceptability of mazindol for the treatment of cocaine 
dependence. The data were presented at the 1990 CPDD meeting and a manuscript has been prepared for 
subnussion to Pharmacol Biochem and Behavior. 



121 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00029-02 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Interaction Between Ethanol and Prostaglandin Synthetase Inhibitors 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: W.B. Pickworth Pharmacologist, BDL, ARCNIDA 

F.R.George Staff Fellow 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

J£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects with histories of moderate alcohol use are studied on the Residential Research Unit to assess the 
effects of ethanol following pretreatment with neither acetaminophen (325, 650, 1300 and 1950 mg) or 
placebo. Acetaminophen is a prostaglandin synthetase inhibitor that has been shown to reduce several 
behavioral and physiologic effects of alcohol in animal studies, alcohol appears to act in part by 
increasing prostaglandin levels. This drug interaction study makes use of our standard procedures for 
assessing abuse potential and performance to evalate the possibility of such antagonistic effects in human 
subjects. This study is conducted in collaboration with the Preclinical Branch. Subject testing has been 
completed. Preliminary analyses indicate that alcohol at this dosage (0.625 gm/kg, taken over 90 mins) 
caused subjective effects (drunk, feel drug, sober, etc) but did not reliably change physiologic or 
performance measures. Pretreatment with acetaminophen did not influence the subjective effects. The 
results are submitted for publication. 



122 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00033-02 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Nicotine Patch: Effects on Smoking Subjective and Physiologic Functions 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: W.B. Pickworth Pharmacologist, BDL. ARC.NIDA 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_X Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

A recently developed nicotine patch will be studied in residential research volunteers. The effect of three 
patches containing 0, 30 and 60 mg will be evaluated on ad lib smoking, subjective effects and 
physiologic measures. The patch will be tested in subjects with and without histories of drug abuse. The 
study is of practical importance in the development of a new therapy for smoking cessation. The subject 
testing phase (n=10) of the experiment has been completed. The results suggest that nicotine patch 
reduced spontaneous smoking in a dose-related fashion. The patch did not cause subject liking and was 
judged to have little abuse potential. The results will be presented at the 1991 CPDD meeting. 



123 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00038-01 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Transcranial Electrostimulation Therapy (TCET) During Smoking Cessation 

Principal Investigators: Cooperating Units 

P.I. W.B. Pickworth Pharmacologist, BDL, ARC.NJDA 

Others: J.E. Henningfield Chief, Clinical Pharmacology Branch 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: 18 Professional: 1.3 Other: .5 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects with histories of cigarette smoking wishing to stop are recruited for this treatment study. 
Subjects are randomly assigned to a treatment group (n=50) which receives small impercatible electric 
current (30 microamps pulsed for 60 min. ) for 1 hr on 5 consecutive days. The sham treated (n=50) 
group are connected to the device but no current is delivered Dependent variables include: cigarettes 
smoked, CO, urinary cotinine, Hatsukami withdrawal score, nicotine craving, mood changes and 
physiologic data (HR, BP). The study has been approved by the IRB and subject recruitment and 
treatment has started. 



124 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00039-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Physiologic and Performance Effects of Ethanol and Pentobarbital and Interactions 
with Indomethacin 

Principal Investigators: Cooperating Units 

P.I. W.B. Pickworth Pharmacologist, BDL, ARC.NIDA 

Others: J.E. Henningfield Chief, Clinical Pharmacology Branch 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .8 Professional: .5 Other: .3 

Check Appropriate Boxes: 

_%, Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects of either sex with histories of alcohol and sedative abuse are studied on the Residential Research 
Unit to assess the effects of indomethacin pretreatment on the performance, physiologic and subjective 
effects of sedative drugs. Animal research suggests that inhibitors of prostaglandin synthesis reduce the 
effects of ethanol but these results have not been confirmed in humans. The protocol has been approved 
by the IRB and subject recruitment and testing has started. 



125 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00013-05 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Archival Data Base Project 

Principal Investigators: Cooperating Units 

P.I. C.A. Haertzen Research Psychologist, BDL, ARC, NIDA 

Others: J.E. Henningfield Chief, Clinical Pharmacology Branch 

W.R. Lange Medical Director. ARC, NIDA 

J.H. Jaffe NIDA 

WE. Weddington Former Staff Fellow 

L. Covi Visiting Scientist 

S. Tiffany Visiting Scientist 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_}£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Data combined into an Admission Data Base consisting of tests obtained on recruitment (Addiction 
Severity Index, SCL-90 Symptom Check List, Shipley IQ) and tests obtained on admission (Diagnostic 
Interview Schedule, Buss-Durkee Hostility, Minnesoat Multiphasic Personality Inventory, and Alcohol 
Related Behavioral Questionnaire) continued to be relevant. Thus, behavioral hostility, as revealed by a 
simulation of the effects of alcohol was related to a diagnosis of alcohol dependence over and above 
prediction afforded by hostility ascribed to a drug free status (see Walter et al., 1990). 

Archival data from the Addiction Research Center Inventory collected in Lexington, Kentucky on 
prisoner heroin addicts for a no-drug condition was analyzed for seasonal effects. Could heroin addicts 
who are frequently depressed show the seasonal pattern typically found for those with seasonal affective 
disorders? Seasonal effects were found, but a summer pattern was found characterized by increased 
sedative-hypnotic effects (tiredness, etc, as measured by the PCAG scale )(see Haertzen, 1991). To 
pursue the question further, the SCL-90 tests obtained on recruitment at the Addiction Research Center in 
Baltimore were analyzed for seasonal effects. Again a significant summer effect was found Depression 
was enhanced. In addition, sleep impairment, as indicated by the items in the Additional Scale, was 
elevated. Psychopathology, as measured at the time of recruitment, may be affected by environmental 
influences or greater behavioral variability in sleeping patterns coincident with summer. 



126 



A significant elevation was also found for SCL-90 Aggression. As a follow-up on this finding 
researchers at the Patuxent prison searched their data base for adverse behavior reports for the year 1990. 
They found an increased incidence in adverse behavior in the summer months, especially June. 

A data base for a questionnaire on cocaine craving designed by Dr. Tiffany was set-up and analyzed. In 
keeping with the results of treatment oriented studies by Drs. Weddington and Covi, a depressive mood 
was correlated with cocaine craving. 

Publications: 

Haertzen, C.A. and Hickey, J.E.: Addiction Research Center Inventory (ARCI): Measurement of 
Euphoria and Other Drug Effects. In Bozarth, M.A. (Ed) Methods of Assessing the Reinforcing 
Properties of Abused Drugs . New York, NY, Springer- Verlag, 1987, pp. 489-524. 

Lange, W.R.. Haertzen, C.A., Hickey, J.E., Snyder, F.R., Dax, E.M., and Jaffe, J.H.: Nitrite inhalants: 
Patterns of abuse in Baltimore and Washington, D.C. Am J Drug Alcohol Abuse 14:29-39, 1988. 

Rose, MR., Brown, B.S., and Haertzen, C.A.: Comparison of the characteristics and functioning of 
cocaine treatment and cocaine research subjects. Am J Drug Alcohol Abuse 15:251-260, 1989. 

Fishbein, D.H., Heming, R.I., Pickworth, W.B., Haertzen, C.A., Hickey, J.E., and Jaffe, J.H.: EEG and 
Brainstem auditory evoked response potentials in adult male drug abusers with self-reported histories of 
aggressive behavior. Biological Psychiatry 595-611,1989. 

Muntaner, C, Nagoshi, C, Jaffe, J.H., Walter, D., Haertzen, C, Fishbein, D.H.: Correlates of self- 
reported early childhood aggression in subjects volunteering for drug studies. Am J Drug Alcohol Abuse 
15:383-402, 1989. 

Weddington, W.W., Brown, B.S., Haertzen, C.A., Cone, E.J., Dax, E.M., Herning, R.I., Michaelson, 
B.S.: Changes in mood, craving, and sleep during acute abstinence reported by male cocaine addicts: A 
controlled, residential study. Archives of General Psychiatry . In press. 

Haertzen, C.A.. Hickey, J.E., Rose, M.R. and Jaffe, J.H.: The relationship between a diagnosis of 
antisocial personality and hostility: Development of an Antisocial Hostility Scale. J Clin Psvchol 
46:679-686, 1990. 

Walter, D., Nagoshi, C, Muntaner, C. & Haertzen, C.A. The prediction of drug dependence from 
expectancy for hostility while intoxicated. International J of Addictions . 25:1 15 1-1 168, 1990. 

Weddington, W.W., Brown, B.S., Haertzen, C.A., Hess, J.M., Mahaffey, J.R., Kolar, A.F. & Jaffe, J.H. 
Comparison of amantadine and desipramine combined with psychotherapy for treatment of cocaine 
dependence. American Journal Drug Alcohol Abuse 17:137-152, 1991. 

Haertzen. C.A. Geophysical variables and behavior: LXV. Seasonal changes in mood in opioid addicts 
on the Addiction Research Center Inventory. Psychological Reports 68:360-362 1991. 



127 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00040-01 BDL 

January 1. 1990 to December 31, 1990 

Title of Project: Opioid Agonist and Antagonist Sensivity in Opiate-Experienced Individuals 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL, NIDA/ARC 

C. Cohen Visiting Scientists, BDL, NIDA/ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

X.Human Subjects _Human Tissues _ Neither 

Minors 

Interviews 

Summary of Work 

Several recent studies have investigated the phenomenon of acute opioid physical dependence or 
antagonist sensivitity in humans. These studies have shown that a mild withdrawal syndrome can be 
precipitated by the opioid antagonist, naloxone, administered 45 minutes to 24 hours after a single dose 
of morphine. These findings are particularly interesting given that the opioid abstinence syndrome has 
traditionally been thought to develop only after prolonged exposure to opiates. However, these studies 
have only used non-dependent opiate-experienced subjects. It is not known whether or to what extent an 
opiate history contributes to the development of acute dependence. If differences in agonist and/or 
antagonist sensitivity are found between opiate-experienced and opiate-naive individuals, it could reflect 
residual tolerance or dependence in opiate-experienced individuals due to their chronic opioid exposure. 
Another interesting possibility is that differences in sensitivity may reflect biological differences between 
opiate-experienced and opiate-naive individuals that predate initial drug use and thus confer some 
vulnerability or predisposition to use or abuse opiate drugs. This research will examine the following 
issues: a) differences in subjective and physiological sensitivity to opioid agonists between opioid- 
experienced and opioid naive subjects, b) differences in acute antagonist-precipitated withdrawal 
(antagonist sensitivity) between these groups, and c) whether any differences in antagonist sensitivity are 
related to dose of the antagonist. 

Subject testing was begun in 1990 and is nearly completed. A preliminary report of data will be 
presented at the 1991 meeting of the Committee on Problems of Drug Dependence. 



128 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00041-01 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Discriminitive Stimulus Effects of Stimulant Drugs 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL, NJJDA/ARC 

C. Cohen Visting Scientist, BDL, NIDA/ARC 



Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

J^Human Subjects _ Human Tissues _ Neither 

Minors 

Interviews 

Summary of Work 

The supply of legally available amphetamines was reduced by about 80% following the 1970 Controlled 
Substances Act, which classified all amphetamine-type stimulants as Schedule II drugs. This void was 
filled by the production of "look-alike" drugs designed to mimic the stimulant effect of amphetamines. 
The main psychoactive agent in many of these preparations is caffeine or caffeine combined with one or 
more sympathomimetic amines, such as ephedrine and phenylopropanolamine (PPA). Individually, 
caffeine, ephedrine, and PPA are relatively safe drugs when taken in moderate or therapeutic doses. 
However, little is known about the behavioral pharmacology of their combined effects in "look-alike" 
stimulants, including their discriminative stimulus effects or potential for abuse when individuals exceed 
the therapeutic dosage regimen in an attempt to achieve amphetamine-like euphoria. 

In a series of studies, we will investigate the subjective, physiological, and discriminative stimulus effects 
of a stimulant that is widely used on a daily basis (caffeine), some that are used clinically (mazindol, 
ephedrine, PPA), and some that have known or potential abuse liability (d-amphetamine, mazindol, 
caffeine/ephedrine/PPA combined "look-alike" stimulants). The knowledge gained from these studies 
will enable us to draw clearer distinctions among stimulants in terms of their behavioral and subjective 
effects. Additionally, these studies will provide seminal information concerning the behavioral 
pharmacology of "look-alike" stimulants, which pose a potential public health risk from a medical and an 
abuse liability standpoint. We plan to begin testing subjects in 1991. 



129 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00042-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Effect of Reinforcement Contingencies on Human Task Performance 

Principal Investigators: Cooperating Units 

P.I.: J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL, NIDA/ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

J^Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Although various psychomotor and cognitive tasks are widely utilized in studies assessing the effects of 
psychoactive drugs on performance, no study, to our knowledge, has systematically examined the effect 
of varying contingencies of reinforcement on human performance. In this context, a reinforcement 
contingency can be defined as an experimental condition in which some monetary rewards is paid upon 
completion of a particular behavior. In most studies, subjects perform tasks with no particular 
consequence for accurate performance or are paid some arbitrary amount for correct responding. This 
lack of attention to the effects of reinforcement contingencies on performance is somewhat surprising 
given the extensive animal literature reporting dramatic effects of manipulating reinforcement conditions 
on schedule-controlled behavior. 

Manipulating reinforcement contingencies on performance is an objective, measureable means of varying 
what is commonly referred to as motivation. Obviously, a person's level of motivation can be an 
important factor in determining the quality and quantity of their performance. The lack of standardizing 
a person's motivational level by holding constant contingencies of reinforcement can result in behavior 
that is subject to uncontrolled variation. A series of studies will examine the effects of manipulating 
reinforcement contingencies on human performance by differentially reinforcing task speed versus task 
accuracy. Subject testing began in 1990, and four separate experiments have been completed (N=21). 
Results indicate that, in general, subjects' behavior has not readily come under the control of the 
reinforcement contingencies. Further studies will be designed to determine what contingency conditions 
reliably control task performance. 



130 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00034-02 DDL 

January 1, 1990 to December 31, 1990 

Title of Project: Opioid Self-Administration in Humans 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL, NJDA/ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The effect of withdrawal states on drug reinforcement and drug self-administration has received little 
systematic attention from drug abuse researchers. This is a critical omission because it is generally 
assumed that humans will seek drugs to alleviate unpleasant or relapsing withdrawal symptoms, thus 
maintaining their state of physical dependence or relapsing after a period of abstinence. This research 
will examine the following issues: (a) the pattern of self-administration when only low doses of opiates 
are available, (b) the effect of opioid antagonist-precipitated withdrawal on opiate self-administration, 
and (c) the relationship between self- administration behavior and subjective drug effects. This research 
should be useful in the development of better methods to predict abuse liability of drugs, because it 
combines the two primary strategies of abuse liability assessment, self-administration and subjective 
effects testing, in a single study. In addition to addressing these important pharmacological-behavioral 
interactions, this research may ultimately result in more effective treatment methods for drug abuse. 
Pilot testing of residential subjects with a history of opioid abuse has been completed, and the main study 
should begin in 1991. 



131 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00026-02 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Assessment of Opioid Agonists and Antagonists 

Principal Investigators: Cooperating Units 

P.I.: J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist. BDL, NIDA/ARC 

E.J. Cone Chief, Chemistry & Drug Metabolism Laboratory 

R.E. Johnson Former Chief. Research Support Branch 

P.J. Fudala Former Deputy Chief, RSB, NJX>A, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Subjects with histories of opioid abuse were studied on the Residential Research Unit to investigate the 
effects of nalmefene, a new investigational opioid antagonist with relatively few agonist effects. This 
research should be useful in determining the possible utility of this long-acting (several days) opioid 
antagonist for the treatment of opioid dependent persons. This research is being conducted in 
collaboration with the Chemistry and Drug Metabolism Laboratory and the Research Support Branch. 
Two studies were planned, the first will assess the abuse liability of nalmefene and the second will 
determine the efficacy of nalmefene to block the subjective and physiological effects of morphine. The 
first study has been completed. Results indicated that nalmefene did not produce typical opiate-like 
abuse liability, but that side effects, such as feelings of agitation and irritability, muscle tension, 
headache, and insomnia may limit its use as possible treatment for opioid dependence. The second study 
was recently completed, and the data are currently being analyzed. A preliminary report will be 
presented at the 1991 meeting of the Committee on Problems of Drug Dependence. 



132 



Abstracts: 

Fudala, P. J., Johnson, RE., Heishman, S.J., Cone, E.J. and Henningfield, J.E. A dose run-up and safety 
evaluation of nalmefene in human volunteers. In L.S. Harris (Ed.), Problems of Drug Dependence 
1989 . NIDA Research Monograph 95 (pp. 451-452). Washington, DC: US Government Printing Office, 
1990. 

Fudala, P.J., Johnson, R.E., Heishman, S.J. and Henningfield, J.E. Evaluation of the abuse liability of 
nalmefene. Clinical Pharmacology and Therapeutics . 49: 167, 1991. 

Publications: 

Fudala, P. J., Heishman, S.J., Henningfield, J.E. and Johnson, R.E. Human pharmacology and abuse 
potential of nalmefene. Clinical Pharmacology and Therapeutics . 49: 300-306, 1991. 



133 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00044-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Psychotropic Properties of Stimulants and Sedatives: Discriminative Properties 

Principal Investigators: Cooperating Units 

P.I.: J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL, NIDA/ARC 

R.J. Lamb Former Staff Fellow 

W.R. Lange Medical Director, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 

Clinical Pharmacology Branch 
Section: 

Institute and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues Neither 

_ Minors 
_ Interviews 

Summary of Work 

In these studies the psychotropic effects of the prototypical stimulant, d-amphetamine, were compared to 
those of another stimulant and to the effects of a sedative and an opioid. These comparisons were 
conducted in subjects with histories of stimulant and either opioid or sedative abuse, and were carried out 
using two different types of procedures simultaneously. The first procedure was a drug discrimination 
task. In the drug discrimination procedure subjects were trained to respond on one lever following 
administration of the training dose of d-amphetamine and on another lever in the absence of d- 
amphetamine. Correct responding was reinforced by money. The second procedure was a traditional 
abuse liability assessment procedure that utilized physiologic and self-report measures. 

To date two studies have been conducted. In the first the effects of amphetamine and hydromorphone 
were compared. In the second the effects of amphetamine, methylphenidate, and diazepam were 
compared. In both studies amphetamine dose-relatedly occasioned d-amphetamine appropriate 
responding. Methylphenidate, also, dose-relatedly occasioned d-amphetamine appropriate responding. 
In contrast neither diazepam nor hydromorphone occasioned d-amphetamine appropriate responding. 
The subjective effects of d-amphetamine and methylphenidate were similar and covaried with their 
discriminative effects, while the subjective effects of diazepam were clearly different. In contrast, the 
only self-report measure that distinguished hydromorphone from d-amphetamine were drug 
identifications. Thus these studies show that drug discrimination procedures can be drug-class specific in 
humans, and that while these discriminative effects can covary with the subjective effects of the drug, the 
discriminative effects of amphetamine under these conditions appear to be controlled in a manner most 
similar to the identification of a drug. 



134 



Abstracts: 

Lamb, R.J. and Henningfield, J.E. (1990) Human d-amphetamine drug discrimination: Testing with d- 
amphetamine and hydromorphone. In: L.S. Harris (Ed.) Problems of Drug Dependence 1989 . NIDA 
Research Monograph 95 (pp. 423-424). Washington, DC: US Government Printing Office. 

Heishman, S.J.. Lange, W.R. and Henningfield, J.E. (1990) Discriminative stimulus effects of d- 
amphetamine. methylphenidate and diazepam in humans. Pharmacology Biochemistry and Behavior . 32: 
1090. 

Heishman, S.J., Lamb, R.J. and Henningfield, J.E. (1990) Discriminative stimulus effects of drugs in 
humans. Stimulants and sedatives. Pharmacology Biochemistry and Behavior . 36: 428. 



Publications: 

Heishman, S.J. and Henningfield, J.E. (1991) Discriminative stimulus effects of d-amphetamine, 
methylphenidate and diazepam in humans. Psvchopharmacologv . in press. 



135 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00031-02 BDL 

January 1. 1990 to December 31. 1990 

Title of Project: Effects of Nicotine in Nonsmokers 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: S.J. Heishman Research Psychologist, BDL. NJDA/ARC 

F.R. Snyder Statistician, NOVA 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.10 Professional: 0.30 Other: 0.8 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Nonsmokers are exposed to nicotine given in the form of nicotine polacrilex gum; preliminary testing 
suggests that this formulation is of low abuse liability and is safe if given according to prescribed 
procedures. Two important experimental questions are addresed in this study. One concerns the further 
evaluation of the effects of nicotine polacrilex gum in nonsmokers to determine the possible effects of 
nicotine on cognitive performance in the absence of pre-existing nicotine dependence. Nicotine enhances 
performance in deprived smokers; however, it remains to be determined if nicotine dependence is a 
precondition for this effect. The second question is of general importance to the understanding of the 
development of drug dependence. Using a model of daily repeated voluntary cumulative dosing, the 
course of possible development of tolerance to the subjective, behavioral and physiologic actions of 
nicotine will be determined Such data cannot be readily obtained with other drugs of abuse, and 
probably not with forms of nicotine known to be of high abuse liability (e.g., cigarettes), but may be 
safely collected following the procedures used in this study. Subject testing is completed (n=12) and data 
are currendy being analyzed. Preliminary results indicate that nicotine did not enhance task performance 
and that tolerance to the effects of nicotine developed for some, but not all, measures. A preliminary 
report of the data was presented at the 1990 meeting of the Committee on Problems of Drug Dependence 
and a more complete report will be made at the 1991 meeting of the American Psychological 
Association. 



136 



Abstracts: 

Heishman. S.J.. Snyder, F.R. and Henningfield. J.E. Effect of repeated nicotine administration in 
nonsmokers. In: Problems of Drug Dependence 1990 . NIDA Research Monograph, in press. 

Heishman, S J., Richards. L.M. and Henningfield, J.E. Effect of nicotine on cognitive and psychomotor 
performance in nonsmokers. Pharmacology Biochemistry & Behavior , in press. 



137 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00045-01 BDL 

January 1. 1990 to December 31. 1990 

Title of Project: Lobeline and Nicotine: Subjective. Physiologic and Kinetic Effects 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.M. Evans Staff Fellow, BDL. NJDA/ARC 

W.B. Pickworth Pharmacologist. BDL, NJDA/ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laborator 
Clinical Pharmacology Branch 

Section: None 

Institute and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Lobeline is used in a variety of over-the-counter aids for smoking cessation but it has never been 
documented in scientific studies to be efficacious as an aid to cessation. Experimental data, however, 
suggests that it may be more effective in alternate formulations and/or presented at higher dose levels. 
Despite the wide spread use of lobeline there are little data on its kinetics or specific dose-related effects 
on human subjective and physiologic responses, and on its effects on cigarette smoking. The major 
purpose of the present study was to determine the safety of lobeline given intravenously over a period of 
5 minutes in doses up to 8 mg. The present study also compared the effects of pretreatment of 
intravenously administered lobeline, placebo and nicotine on physiologic effects, subjective effects and 
kinetics. In addition, the effects of these drugs on cigarette smoking behavior was also evaluated. We 
have recently completed this study and are currently writing up the report. Four subjects completed the 
full dose-range of lobeline and nicotine with no adverse effects on either cardiovascular measures (ECG. 
blood pressure, heart rate) or other subjective effects. These results suggest that lobeline is safe even 
when given as a 5 min intravenous injection up to doses of 8 mg. Inspection of the subjective effects data 
suggest that doses administered were not producing any effects similar to standard drugs of abuse. 
Preliminary results also indicate that the doses of lobeline and nicotine administered using the injection 
procedure in the present study did not show any decreases on cigarette smoking behavior. At this time, 
plasma samples have yet to be analyzed so there are no kinetic data. 



138 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00046-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Arterial Kinetics of Smoked Cocaine 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.M. Evans Staff Fellow, ARC 

E.J. Cone Chief, CDM Laboratory 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_2L Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Currently, smoked cocaine or "crack" is thought to have greater addictive effects than other routes of 
cocaine administration, including intravenous administration. Surprisingly, despite the fact that smoked 
cocaine represents one of the most serious medical and social challenges to our society today there are no 
data from humans on the levels of cocaine that actually reach the arterial blood stream enroute to the 
brain. 

The purpose of this study is to directly compare the effects of smoked cocaine to those of intravenously 
administered cocaine primarily on the kinetics of cocaine in both arterial and venous blood. Standard 
cardiovascular measures and subjective effects will be evaluated concurrendy with the arterial and 
venous blood draws. Subjects will have current histories of using cocaine by both routes of 
administration. Subjects will be exposed to 3 doses of smoked cocaine or "crack" and 3 doses of 
intravenous cocaine. 

To date we have obtained the IND to administer smoked cocaine. We have also met several times with 
the anesthesiologists to review the protocol and technical details with the study nurses. We are planning 
to recruit our first subject mid-May. 



139 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00047-01 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Outpatient Drug Discrimination 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: S.M. Evans Staff Fellow, BDL, NIDA/ARC 

C.E. Johanson Collaborator, USUHS 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The purpose of this study was to determine if normal volunteers could be trained to discriminate 
therapeutic doses of antihistamines (e.g., tripelennamine and diphenhydramine) from placebo and to 
evaluate the dose-relationship and pharmacological specificity of the discrimination. This protocol is 
currently under review by the fRB. 



140 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00006-04 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Triazolam Self-Administration: Effects of Yohimbine Pretreatment 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: JR. Roache Former Staff Fellow, BDL. NIDA, ARC 

R. A. Meisch Former Staff Fellow, BDL, NIDA, ARC 

S. A. Klein Former Staff FeUow, BDL. NBDA, ARC 

J.H. Jaffe Former Director, NJDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.70 Professional: 2.00 Other: 5.00 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The purpose of this study was to examine the effects of yohimbine pretreatment on the self- 
administration of triazolam in subjects with histories of sedative abuse. Two issues of relevance to the 
behavioral pharmacology of drug abuse are being addressed: the first involved the development of 
procedures to measure sedative/anxiolytic drug self-administration: and, the second was to examine the 
effects of yohimbine pretreatment on triazolam self-administration. It is of basic theoretical, as well as 
clinical, interest to define methods to detect the effects of one drug on the self-administration of another 
drug. In addition, yohimbine has been shown to produce neuroendocrine changes and subjective mood 
states in humans which resemble anxiety. Thus, this study could provide important information related to 
hypotheses of drug abuse which ivolve psychiatric vulnerability factors. 

Data collection has been completed and a manuscript has been submitted for publication. 



141 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00009-06 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Effects of Drugs on Cigarette Smoking and Responses to Nicotine 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R. Nemeth-Coslett Former Staff Fellow, BDL, NJDA, ARC 

F.C. Davis Nurse, FSK 

A.H. Sampson Nurse, FSK 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.23 Professional: 0.03 Other: 0.20* 

Check Appropriate Boxes: 

X. Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Initial studies were conducted in the physical facilities of the Behavioral Pharmacology Research Unit at 
Johns Hopkins University medical School in collaboration with Dr. R.R. Griffith which provided some 
research support. For example, multiple measures of cigarette smoking, subjective effect, and 
physiologic effect were collected during ad libitum smoking sessions in normal volunteers following 
administration of mecamylamine, naloxone, or marijuana. 

Presently, basic measures of cigarette smoking are being collected from all subjects on the Clinical 
Research Unit and data analyses have begun. This database-type of study appears to be providing the 
opportunity to quantitate the effects of a wide range of variables on cigarette smoking (i.e., atropine 
administration, cocaine withdrawal, buprenorphine administration, and passive tobacco smoke exposure). 



142 



Publications 

Nemeth-Coslett, R. and Griffiths, R.R.: Naloxone does not affect cigarette smoking. 
Psvchopharmacologv . 89:261-264, 1986. 

Nemeth-Coslett. R., Henningfield, J.E., O'Keeffe, M.K. and Griffiths. R.R.: Effects of mecamylamine on 
human cigarette smoking and subjective ratings. Psvchopharmacologv , 88:420-425, 1986. 

Nemeth-Coslett, R., Henningfield, J.E., O'Keeffe, M.K. and Griffiths, R.R.: Effects of marijuana on 
human cigarette smoking and physiologic changes and subjective responses. Pharmacol Biochem Behav 
25:659-665, 1986. 

Nemeth-Coslett, R., Henningfield, J.E., O'Keeffe, M.K. and Griffiths, R.R.: Nicotine gum: Dose-related 
effects on cigarette smoking and subjective ratings. Psvchopharmacologv 92:424-430, 1987. 

Rose. J.E., Sampson, A.. Levin, E.D. and Henningfield, J.E.: Mecamylamine increases nicotine 
preference and attenuates nicotine discrimination. Pharmacol Biochem Behav . In press. 



143 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00010-06 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Behavioral and Pharmacologic Factors in Nicotine Replacement for Tobacco 
Dependence 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief. Clinical Pharmacology Branch 

Others: W.B. Pickworth Pharmacologist, BDL, NIDA, ARC 

C. Cohen Visiting Scientist, BDL, NJDA, ARC 

Eric Simmons Coppin State College 

Alex Radzius Research Assistant, BDL, NJDA. ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.50 Professional: 0.15 Other: 0.35 

Check Appropriate Boxes: 

X_ Human Subjects Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Nicotine polacrilex (chewing gum) has been under investigation as a replacement for tobacco-delivered 
nicotine and also as a convenient drug administration modality which provides a model of more general 
interest for drug dependence researchers. For example, nicotine gum was employed in our initial studies 
to examine the capabilities of this Laboratory's recently established performance and electrophysiologic 
assessment approaches for evaluating drug effects. The course of research conducted using this 
preparation has been determined by the priorities of the ARC and the Chief of the Biology of 
Dependence Laboratory. These studies have included the following: (1) Effects of nicotine gum 
replacement on cigarette smoking and tobacco smoke exposure; (2) Pharmacodynamic effects of nicotine 
gum compared to other routes of nicotine administration; (3) Abuse liability of nicotine gum; (4) Dose- 
related effects on subjective, behavioral, and physiologi variables, including studies of the factors which 
may affect the functional dose, such as chewing and swallowing rates; (5) Effects of nicotine gum 
admiiustration on learning and performance in non-smokers; and (6) Role of oral pH in nicotine 
absorption. 



144 



Publications 

Nemeth-Coslett. R.. Benowitz. N.L., Robinson, N. and Henningfleld. J.E.: Nicotine Gum: Chew rate, 
subjective effects and plasma nicotine. Pharmacol Biochem Behav 29:747-751, 1988. 

Pickworth, W.B., Heming, R.I. and Henningfieid. J.E.: Electroencephalographic effects of nicotine gum 
in humans. Pharmacol Biochem Behav 25:879-882. 1986. 

Jasinski, D.R and Henningfieid, J.E.: Conceptual Basis of Replacement Therapies for Chemical 
Dependence. In Pomerleau. O.F., Pomerleau, C.S., Fagrstrom, K.O., Henningfieid, J.E. and Hughes. JR. 
(Eds): Nicotine Replacement: A Critical Evaluation . New York, NY, Alan R. Liss, 1988, pp. 13-34. 

Waranch, H.R., Henningfieid. J.E. and Edmunds, M: Letter to the editor: Elimination of nicotine gum 
use following successful replacement therapy for cigarette smoking. Lancet January 2-9:49-50, 1988. 

Snyder, F.R. and Henningfieid, J.E.: Effects of acute nicotine deprivation and administration: 
Assessment on computerized performance tasks. Psvchopharmacologv . In press. 

Pickworth, W.B.. Heming, R.I. and Henningfieid, J.E.: Mecamylamine reduces some EEG effects of 
nicotine chewing gum in humans. Pharmacol Biochem Behav 30:149-153, 1988. 

Pomerleau. OF., Pomerleau, C.S., Fagerstrom, K.O., Henningfieid. J.E. and Hughes, JR.: (Eds.): 
Nicotine Replacement: A Critical Evaluation . New York, NY, Alan R. Liss, 1988. 

Jarvik. ME. and Henningfieid. J.E. Pharmacological adjuncts for the treatment of nicotine dependence. 
In: J.D. Slade and C.T. Orleans (eds.) Nicotine Addiction: Principles and Management . Oxford 
University Press, in press. 

Cone, E.J. and Henningfieid. J.E. Premier smokeless cigarettes' can be used to deliver crack. Journal of 
the American Medical Association 261(1 ):41. 1989. 

Rose, J.E., Sampson. A., Levin, E.D. and Henningfieid, J.E. Mecamylamine increases nicotine 
preference and attenuates nicotine discrimination. Pharmacology Biochemistry and Behavior 32(4):933- 
938, 1989. 

Henningfieid, J.E. and Woodson, P.P. Dose-related actions of nicotine on behavior and physiology: 
Review and implications for replacement therapy for nicotine dependence. Journal of Substance Abuse 
1:301-317, 1989. 

Henningfieid, J.E., London, E.D. and Benowitz, N.L. Arterial-venous differences in plasma 
concentrations of nicotine after cigarette smoking. Journal of the American Medical Association 
263:2049-2050, 1990. 

Boren, J.J., Stitzer, M.L. and Henningfieid, J.E. Preference among research cigarettes with varying 
nicotine yields. Pharmacology Biochemistry and Behavior 36:191-193. 1990. 

Henningfieid, J.E., Radzius, A., Cooper, T.M. and Clayton, R.R. Drinking coffee and carbonated 
beverages blocks absorption of nicotine from nicotine polacrilex gum. Journal of the American Medical 
Association , 264:1560-1564, 1990. 

Henningfieid. J.E., Radzius, A.. Cooper, T.M. and Clayton, R.R. Oral pH: A factor in the treatment of 
nicotine addiction. Journal of the American Dental Association , under review. 



145 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00024-03 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Opioid Self-Administration: Humans Compared to Animals 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R.J. Lamb UMDNJ 

S.R. Goldberg Chief, Preclinical Branch 

J.L. Katz Chief, Psychobiology Laboratory, ARC 

C.W. Schindler Staff Fellow, NJDA, ARC 

R.A. Meisch Visiting Scientist, U of Tex, Houston 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.15 Professional: 0.10 Other: 0.05 

Check Appropriate Boxes: 

_^£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Studies with animals have shown that stimuli associated with drug delivery can come to function as 
variables that partially control drug-seeking behavior and the likelihood of resumption (i.e., relapse) to 
such behavior, even in the absence of the drug. Analogous research strategies are being used to assess 
the generality of these findings to human subjects. In addition, these procedures provide data on the 
degree of correspondence between self-reported drug effects and drug seeking behavior. The human 
studies have produced a number of interesting results. When the consequences of varying the dose of 
morphine available on self-administration, physiological effects, and self-reported effects were examined, 
it was found that low doses of morphine (3.75 mg) maintained rates of responding above placebo and 
constricted pupillary diameter, but did not reliably alter the self-reports of the subjects, indicating a 
dissociation between the subjective effects of morphine and morphine's reinforcing properties. Another 
study evaluated the role of a stimuli paired with drug administration on the maintenance of responding. 
Initial results suggested that the stimuli were of less importance than in an analogous study with animals, 
as well as in somewhat similar study of cocaine self- administration by humans. 



146 



Publication 

Lamb. R.J., Preston, K.L., Henningfield, J.E., Schindler, C.W., Meisch, R.A., Davis, F., Katz, J.L. and 
Goldberg, S.R.: The Reinforcing and Subjective Effects of Morphine in Post- Addicts: A Dose-response 
Study. J Pharm Exp Ther . Submitted. 



147 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00007-05 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Effects of Commonly Used Drugs on Behavioral Performance in Normal Subjects 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: P.P. Woodson Former Staff Fellow, BDL, NIDA, ARC 

J.D. Roache Former Staff Fellow, BDL, NIDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.45 Professional: 0.45 Other: 1.00 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The possible adverse effects on performance of an antihistamine and alcohol were evaluated in non- 
residential subjects without histories of drug abuse, other than cigarette smoking. The study involved the 
use of strategies recommended by the Joint Triservices Working Group (Army Contract) to assess 
behavioral (i.e., cognitive) performance. Measures included the standard Army Performance Assessment 
Battery (PAB), prototypic portions of the Unified Triservices Battery (UTC PAB), critical flicker fusion, 
and mood, as well as cardiovascular and other basis physiologic variables. 

Preliminary analysis of data from the first study suggest that alcohol and chlorpheneramine produced 
dose-related effects on several self-report measures and mixed effects on performance across measures. 
These initial results suggest that the PAB is less sensitive compared to the Digit Symbol Substitution 
Task with respect to the level of performance disruption by alcohol or chlorpheramine. 

Subsequently, a new protocol compared a non-centrally acting antihistamine (terfenadine) to a centrally 
acting one (diphenhydramine) as well as to the benzodiazepine, triazolam. Data collected has been 
completed and the results are presendy being analyzed to prepare a report for publication. This was the 
final study in the series of those conducted in collaboration with the Joint Triservices Working Group 
(Army Contract). 



148 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00030-02 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Passive Tobacco Smoke: Nicotine Absorption, Subjective Effects and Performance 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: P.P. Woodson Former Staff Fellow 

J.D. Roache Former Staff Fellow 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.35 Professional: 0.15 Other: 0.20 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Volunteers were tested to determine the effects of exposure to ambient tobacco smoke, generated by a 
cigarette smoking machine, on standard measures of subjective and physiologic effect as well as on 
performance. It was hoped that the use of the performance battery included in this study would provide a 
quantitative assay by which to determine if various ambient levels of tobacco smoke can produce dose- 
dependent effects on performance and physiology which are comparable to those observed with cigarette 
smoking. Initial research demonstrated the safety and reliability of the procedures for inducing passive 
tobacco smoke exposure. Data collection has been completed and a report for publication is in 
preparation. 



149 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00004-05 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Comparative Self- Administration (Monkeys and Human): Nicotine and Cocaine 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R.J. Lamb Former Staff Fellow 

S.R. Goldberg Chief, Preclinical Branch 

C.W. Schindler Staff Fellow, Preclinical Branch 

Lab/Branch: Biology of Dependence and abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: Professional: Other: 

Check Appropriate Boxes: 

^X Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

This was a collaborative project with the BPL in which the human research was conducted on the 
Residential Research Unit and parallel animal studies were conducted in the BPL. The use of the self- 
administration (SA) study paradigm permitted an assessment of the relative contribution of 
environmental and pharmacologic factors to the self-administration of drugs. Parallel comparative 
studies in squirrel monkeys and humans in which subjects are given the opportunity to self-administer 
comparable doses of cocaine and nicotine under similar behavioral schedules and experimental 
conditions also provide a means to assess the generality of biological variables influencing drug SA. This 
research has shown that responding is maintained in human subjects. The stimuli that are associated with 
injections of cocaine develop conditioned reinforcing effects in the humans in a manner similar to the 
manner in which these effects develop in squirrel monkeys. These studies have also demonstrated that a 
research strategy employing drug SA in human subjects can yield all of the important information of 
single-dose studies, and also, provide information on the direct reinforcing effects of the compound 
which may be compared to the large base of animal drug SA. Data collection has been completed and 
need only to undergo final analyses before publication. 



150 



Publications 

Goldberg. S.R. and Henningfield. J.E.: Reinforcing effect of nicotine in humans and experimental 
animals responding under intermittent schedules of i.v. drug injection. Pharmacol Biochem Behav 
30:227-234, 1988. 

Henningfield, J.E.. Nemeth-Coslett, R., Katz, J.L. and Goldberg, S.R. Intravenous cocaine self- 
administration by human volunteers: Second-order schedules of reinforcement. In: Harris, L.S. (ed.): 
NIDA Research Monograph 76 . Washington, D.C.: U.S. Government Printing Office, 1987, pp. 266-273. 

Henningfield. IE. and Goldberg, S.R. Pharmacological determinants of tobacco self-administration by 
humans. Pharmacol Biochem Behav 30:221-226, 1988. 



151 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00025-03 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Acquisition of Dependence to Cigarettes and Smokeless Tobacco 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: E.J. Cone Chief, CDM, NIDA, ARC 

A. Radzius Research Assistant, BDL, NIDA, ARC 

K.O. Fagerstrom Sweden 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: Pharmacia Leo therapeutics AB, Helsingborg, Sweden 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 50 Professional: 0.25 Other: 0.25 

Check Appropriate Boxes: 

_2£ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Questionnaires were given to populations of experienced cigarette and/or smokeless tobacco users (785 
responses), and to a population which included persons who had never used tobacco (496 responses). 
The purpose of the questionnaires was to determine changes in the amount of tobacco products consumed 
as a function of time and to assess the level of nicotine dependence, as measured by the Fagerstrom 
Tolerance FQuestionnaire (FTQ). Findings that have emerged from initial analysis of the first population 
include the following: (1) Smokeless Tobacco use begins about one year earlier than cigarette use (15.5 
vs 16.3); (2) Males begin smoking about one year earlier than females; (3) Tobacco consumption 
increased over time (i.e., dose graduation); (4) The dose escalation was negatively accelerated with no 
difference between sexes; (5) Age of starting smoking is negatively correlated with the age of quitting 
and also with predicted FTQ scores after the same number of years of smoking; (6) Four of 8 questions 
on FTQ scale are correlated with total FTQ score. Analyses in progress are: (1) Analysis of brands 
smoked; (2) Prediction of dependence based on the amount of tobacco product consumbed at some early 
point in history; and, (3) Analysis of the data from the 496 response population. These data need only to 
undergo final analyses before publication. 



152 



Publications 

Henningfield. J.E., Clayton. R. and Pollin, W. The involvement of tobacco in alcoholism and illicit drug 
use. British Journal of Addiction 85:279-292. 1990. 

Henningfield. J.E., Cohen, C. and Giovino, G.A. Can genetic constitution affect the 'objective' diagnosis 
of nicotine dependence? American Journal of Public Health , 80:1040-1041. 1990. 

Brigham. J.. Henningfield, J.E. and Stitzer, M.L. Smoking relapse: A review. International Journal of the 
Addictions , in press. 

Henningfield. J.E., Cohen, C. and Slade, J.D. Is nicotine more addictive than cocaine? British Journal of 
Addictions , in press. 



153 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00014-03 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Cholinergic Agonists and Antagonists 

Principal Investigators: Cooperating Units 

P.I.: J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R.I. Herning Staff Fellow. Etiology Branch 

W.B. Pickworth Pharmacologist, BDL. NIDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: Etiology Branch 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: 70 Professional: 0.20 Other: 0.50 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Ten male volunteers without histories of drug abuse, except for cigarette smoking, were tested to assess 
the possible adverse performance effects of a cholinomimetic and a cholinergic antagonist, each given 
alone and in combination. A dose run-up procedure was employed in which physostigmine was 
administered i.v. in an ascending dose series (0.25, 0.5, 1.0, 1.5, and 2.0 mg) first alone, then following 
pretreatment with 5.0 or 10. mg of methscopolamine, a peripherally active antagonist. 
Methscopolamine was given to assess the degree to which peripheral blockade reduced physiological 
effects and/or performance impairment. The Army Performance Assessment Battery (PAB), including 
components of the Triservices PAB. was used to evaluate behavioral performance. Preliminary analyses 
are ongoing. 



154 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00012-07 BDL 

January 1. 1990 to December 31. 1990 

Title of Project: Factors Influencing Behavioral and Physiologic Response to Opioids (Mu Project) 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: E.J. Cone Chief, CDM. NTDA, ARC 

ST. Higgins Univ. of Vermont 

K. L. Preston BPRU, JHU 

J.H. Jaffe Former Director, NIDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: Biology of Vulnerability, Chemistry and Drug Metabolism Laboratory 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.90 Professional: 0.40 Other: 0.50 

Check Appropriate Boxes: 

_X Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Following from the observations that post-addicts and non-opioid users are differentially sensitive to 
opioids, and perhaps even respond qualitatively differently, and the possibility that such differences 
either predispose certain persons to opioid abuse and/or contribute to relapse, this study was conducted to 
experimentally examine such population differences in response to mu and kappa opioids. Prominent 
measures included discrimination thresholds of behavioral effects, physiologic responses, and 
neuroendocrine response. Post-addict and opioid-naive subjects were intended to be separately tested for 
comparison. Initial phase testing involving post-addict volunteers was completed, however, changes in 
priorities resulted in the termintion of the protocol before opioid-naive subjects were tested. The final 
report has been submitted for publication. 



155 



Publication: 

Higgins, ST., Preston. K.L., Cone, E.J., Henningfield, J.E. and Jaffe. J.H. Behavioral, Physiological, and 
Hormonal Effects of a Naloxone Challenge Following Acute Morphine Pretreatment in Humans. In 
Harris, L.S. (Ed.), NEDA Research Monograph 76 . Washington, D.C., U.S. Government Printing Office, 
1987, pp. 266-273. Manuscript submitted. 



156 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00005-05 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Abuse Liability of Smokeless Tobacco Products 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: A. Radzius Research Assistant, BDL, NJDA, ARC 

E.J. Cone Chief, CDM, NJDA, ARC 

N.L. Benowitz Collaborator. Univ. of California 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: Chemistry and Drug metabolism Laboratory 

Division of Clinical Pharmacology and Experimental Therapeutics 
University of California, San Francisco 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.37 Professional: 0.07 Other: 0.30 

Check Appropriate Boxes: 

_£ Human Subjects _ Human Tissues _ Neither 

Minors 
_ Interviews 

Summary of Work 

In two studies, tobacco users were tested with a commercially available smokeless tobacco product (i.e., 
pouches of snuff), and with a smokeless cigarette through which air is sucked to inhale vaporized 
nicotine. Standardized methods of abuse liability assessment were used. 

The smokeless tobacco study consisted of two phases. The first evaluated the effects of dose and the 
possibility that rate of expectoration would alter nicotine extraction and effects. Dose-related changes 
were found in the magnitude and duration of action of measures such as reduction in urge to smoke and 
strength of effects observed. The second phase evaluated the relationship of the effects observed to 
plasma levels of nicotine; these were found to be closely related to the dose administered, thus 
confirming the reliability of this system of nicotine delivery. The study was smokeless cigarettes 
indicated similar dose-related effects as those found with the commercial tobacco products; nicotine 
levels were negligible, suggesting the possibility that this route of nicotine administration may produce 
effects mediated by its peripheral stimulus properties which resemble those of smoking cigarettes. 
Testing has been completed. 



157 



Publications 

Henningfield, J.E. How tobacco produces drug dependence. In: J.K. Ockene (ed.). The Proceedings of 
the World Congress on the Pharmacologic Treatment of Tobacco Dependence . Cambridge, MA: 
Institute for the Study of Smoking Behavior and Policy, pp. 19-31, 1986. 

Cullen. J.W., Blot, W., Henningfield, J.E., Boyd, g., Mecklenberg, R. and Massey, M.M. Health 
consequences of using smokeless tobacco: Summary of the Advisory Committee's Report to the Surgeon 
General. Public Health Reports 101:355-373, 1986. 

Connolly, G.N., Winn, D.M., Hecht, S.S., Henningfield, J.E., Walker, B. and Hoffman, D. The re- 
emergence of smokeless tobacco. N Eng Med 314:1020-1027, 1986. 

Glover, ED., Schroeder, K.L., Henningfield, J.E., Severson, H.H. and Christen, A.G. An interpretative 
review of smokeless tobacco research in the United States: Part I. Journal of Drug Education 18(4):285- 
310, 1988. 

Glover. E.D., Schroeder, K.L.. Henningfield, J.E., Severson, H.H. and Christen, AG. An interpretative 
review of smokeless tobacco research in the United States: Part II. Journal of Drug Education 19(1): 1-19. 
1989. 



158 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00032-02 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Dopaminergic lessions and subjective effects of methylphenidate 

Principal Investigators: Cooperating Units 

P.I. G.R. Uhl Visiting Scientist, MNL, NJJDA, ARC 

Others: M.J. Kuhar Chief, NB. NJDA. ARC 

J.E. Henningfield Chief, CPB, NBDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.30 Professional: 0.10 Other: 0.20 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The purpose of this research study is to examine whether the effect of the drug "Methylephenidate" that 
has been used in the therapy of Parkinson's disease is different in patients with Parkinson's disease 
compared with individuals without this disease. The study will test whether differences in feeling that 
these drugs can induce in normal individuals may or may not be present in patients with Parkinson's 
disease. Preliminary testing was initiated in 1989, the final phase of this study is now underway. 



159 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00035-02 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Wby do substance abusers seek help? What are their worries about that help? 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: RE. Johnson Former Chief, RSB, NJDA, ARC 

D. Brooke Visiting Scientist, RSB, NJDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.20 Professional: 0.10 Other: 0.10 

Check Appropriate Boxes: 

_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
X_ Interviews 

Summary of Work 

A survey of ARC research subjects was conducted to investigate the reasons that people seek treatment, 
and what their worries about that treatment are. The intent of this protocol is to obtain answers to these 
questions which will enable us to make it easier for people to seek help. Subjects were asked to fill out 
two questionnaires and a cover sheet on their past experience of seeking help. A preliminary manuscript 
has been developed and will be submitted for publication. 



160 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00011-05 BDL 

January 1, 1990 to December 31. 1990 

Title of Project: Physiologic Dependence to Tobacco: Cigarette Withdrawal and Nicotine Substitution 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R. Nemeth-Coslett Former Staff Fellow, BDL, NJDA, ARC 

R. Herning Staff Fellow, EB, NIDA, ARC 

W.B. Pickworth Pharmacologist, BDL, NIDA, ARC 

W.R. Lange Medical Director, NIDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Ginical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .80 Professional: 55 Other: .25 

Check Appropriate Boxes: 

^X Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Heavy tobacco users, otherwise without drug abuse histories, were studied on the Residential Research 
Unit. In the withdrawal study, subjects were assessed for nicotine and cotinine, general cardiovascular 
functioning, passive EEG and evoked cortical potential, and caloric intake, during 10 days of cigarette 
deprivation and when smoking resumed. In the substitution phase of the study, subjects were tested 
during alternating cycles of 4 days smoking and 3 days abstinence. In this phase, subjects were similarly 
assessed as described above, but on days in which they were not permitted to smoke, they were given 
pieces of gum to chew 12 times per day at one hour intervals: the gum contained either 0, 2 or 4 mg of 
nicotine. We found that an orderly withdrawal emerged. It included impaired performance, which did 
not recover within the ten days of abstinence, but did recover when cigarette smoking resumed. Nicotine 
gum reversed major signs of tobacco withdrawal, confirming that the withdrawal was nicotine specific. 
This effect was dose-related, e.g., 4 mg gum restored performance to baseline levels, whereas 2 mg gum 
only partially restored performance. Placebo gum use was accompanied by withdrawal. Together, these 
results confirm that nicotine replacement can be a viable mode of alleviation of the tobacco withdrawal 
syndrome, but is of little benefit in reducing desire to smoke (which appears to by pharmaccologically 
related to abstinence but appears readily elicited by environmental stimuli). 



161 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00008-04 BDL 

January 1, 1990 to December 31, 1990 

Title of Project: Behavioral Performance and Physiologic Effects of Drugs: Atropine and Diazepam 

Principal Investigators: Cooperating Units 

P.I. J.E. Henningfield Chief, Clinical Pharmacology Branch 

Others: R.J. Lamb Former Staff Fellow, NIDA, ARC 

S.T. Higgins Former Staff Fellow, NIDA, ARC 

R.I. Herning Staff Fellow, EB, NIDA, ARC 

W.B. Pickworth Pharmacologist, BDL, NIDA, ARC 

F.R. Snyder Statistician 

W.R. Lange Medical Director, NIDA, ARC 

Lab/Branch: Biology of Dependence and Abuse Potential Assessment Laboratory 
Clinical Pharmacology Branch 

Section: Etiology Branch 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .10 Professional: 5 Other: 5 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Studies were conducted to assess the effects of the drugs on performance at various tasks. An additional 
aspect of this research was to identify possible electrophysiological effects. Atropine and diazepam were 
found to produce dose-related effects on a variety of measures of subjective response as well as 
performance on the computerized Performance Assessment Battery (PAB). 

These studies have been completed and data analyses are underway. Preliminary analysis of the results 
from the study on diazepam indicate that most measures were affected in an orderly time and dose- 
related manner. Most measures were surprisingly insensitive, however, and significant effects were often 
not seen until the administration of the highest dose of diazepam (40 mg). the Army developed measures 
did not appear to be more sensitive than traditional measures (e.g., DSST). 



162 



Publications: 

Higgins. ST., Woodward, B.M. and Henningfield, J.E. Effects of atropine on the repeated acquisition 
and performance of response sequences in humans. Journal of the Experimental Analysis of Behavior 
51:5-15, 1989. 

Higgins, ST., Lamb, R.J. and Henningfield. J.E. Dose-related effects of atropine on behavioral and 
physiologic responses in humans. Pharmacology Biochemistry and Behavio r 34(2):303-31 1 , 1989. 



163 



2. Chemistry and Drug Metabolism Laboratory - Edward J. Cone, Ph.D. 

The Laboratory of Chemistry and Drug Metabolism performs chemical, pharmacokinetic, metabolic and 
pharmacodynamic research with human subjects related to the chemistry of substance abuse. Presently, 
studies are underway to further delineate the pharmacokinetic and pharmacodynamic profile of 
marijuana, opiates and methamphetamine. The focus of these studies is the exploration of the 
relationship between drug levels in various body fluids to behavior, performance and physiological 
effects. An additional focus is the evaluation of the usefulness of unusual body fluids or tissues, e.g., 
saliva, nails and hair for drug detection. Our initial findings indicate that each body fluid or tissue 
provides a unique but slighdy different historical record of drug exposure. It is important in the 
diagnosis, treatment and prevention of drug abuse that we have an understanding of the information 
provided by drug tests on various body fluids and the relationship of these tests to drug-induced effects. 
Also, the risk of unknowing drug exposure, e.g., "crack" smoke and methamphetamine smoke, is being 
evaluated. The laboratory performs basic research in the area of chemical methodology development: 
new methods must be developed as new drugs appear in the illicit drug market and as new and important 
metabolites are identified in metabolic studies. In related studies, the validity of commercial test 
methods presently used in employment drug testing are being evaluated for precision and accuracy with 
clinical specimens collected under highly controlled conditions. 

Summary of Ongoing Research 

Specific research projects which were actively pursued in 1990 are briefly summarized below. Only 
those studies for which personnel from this laboratory were the principal investigators are discussed. 

A. The Pharmacokinetics and Pharmacodynamics of Opiate Analgesics. 

The goals of this study include: 1 ) the evaluation of the usefulness of a heroin "marker" as a means of 
detecting heroin addicts in urine and saliva of subjects after heroin abuse; 2) determination of the 
relationship between plasma levels and saliva levels of active drug or metabolite and pharmacologic 
effects: 3) the use of saliva as a screening media for opiates; and 4) validity assessment studies of 
commercial drug assays for opiates. 

B. Studies on the Validity of Drug Testing Methodology. 

The goal of this study is to compare test results of commercial screening assays for drugs in urine with 
test results obtained by gas chromatograpby/mass spectrometry (GC/MS). Standard specimen sets 
utilized in these studies consist of clinical drug specimens collected under highly controlled conditions 
following drug administration and "spiked" standards at known concentration. A complete validity 
assessment of opiate assays was completed utilizing clinical drug specimens collected following heroin, 
morphine, codeine, hydromorphone, hydrocodone, buprenorphine, oxymorphone and oxycodone 
administration. 

C. Drug Assay Development Studies on Drugs of Abuse. 

The aim of this ongoing project is to develop specific, sensitive and reliable assays for drugs of abuse in a 
variety of biological media. For example, test methodology was developed for the detection of opiates in 
hair. This assay was used to study the appearance of morphine and codeine in human facial hair after 
controlled dosing. This assay provided the first documented evidence of the time period required for an 
administered opiate to appear in hair. Work also continues on the refinement of an assay for the 
simultaneous assay of cocaine and metabolites in body fluids. Another assay is currently under 
development for the determination of buprenorphine in blood, saliva and urine. Buprenorphine is a 
promising new drug for the treatment of opiate and cocaine addiction. Other assays also are developed 
for support of ongoing pharmacokinetic and pharmacodynamic studies. 



164 



D. Buprenorphine Pharmacodynamics. 

Buprenorphine is an opioid partial agonist which shows promise as a treatment agent for heroin and 
cocaine addiction. Although buprenorphine has limited bioavailability by the oral route of 
administration, it is effective by the sublingual route. Current studies are underway to determine its 
bioavailability by the sublingual and buccal route. Concurrent behavioral and physiological effects will 
be measured for correlation with blood levels. Urine will be tested for buprenorphine and metabolite 
content. The detection period for buprenorphine in urine and saliva will be determined. In addition , 
following chronic buprenorphine dosing, the blood levels of drug and metabolite will be determined in 
order to evaluate the importance of accumulation of drug and active metabolites. 

E. Fast Action Dynamics of Marijuana Smoking. 

The immediate effects of smoking marijuana on behavior and performance will be evaluated in this 
study. Behavioral and physiological measures will be collected before, during and after smoking. Blood 
and saliva samples will be collected concurrendy and will be analyzed for tetrahydrocannabinol and 
metabolite content as well as selected hormones. The study is designed to evaluate the mechanistic and 
functional effects of smoking marijuana in human subjects. The study will have a unique focus on the 
early changes that occur in the physiology, behavior and the neuroendocrine system during the smoking 
of marijuana cigarettes. 

F. Passive Inhalation of Drugs of Abuse. 

When drugs of abuse are smoked, e.g., marijuana, cocaine, heroin, phencyclidine and methamphetamine, 
some of the volatile material is released into the atmosphere. Depending on the local environment, 
bystanders may be exposed to small doses of the drug and its pyrolyzed breakdown materials. 
Laboratory methods are being developed to experimentally simulate an atmosphere of drug smoke and 
means of withdrawing air samples for chemical analysis. These methods will be used to assess the 
potential hazards of passive inhalation of drugs of abuse. 

G. Methamphetamine Pharmacodynamics. 

Methamphetamine is a stimulant with effects similar to cocaine. Historically, it has a history of abuse 
both as a licit and illicit drug. Presendy, there is concern that a new form of methamphetamine. "ice", 
may be abused in the same manner as the smokeable form of cocaine, i.e., "crack". Methods are under 
development to study the effects of this new form of methamphetamine. The pharmacokinetic profile, 
abuse liability and chemistry of "ice" will be evaluated. 



165 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00002-05 CDM 

January 1, 1990 to December 31, 1990 

Title of Project: Validity Studies of Commercial Drug Screening Assays 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief CDM, ARC. NIDA 

Others D. Darwin Chemist ARC, NIDA 

S. Dickerson Lab Tech ARC, NEDA 

T. Maguire Chemist ARC, NIDA 

B. Holicky Nurse ARC, NIDA 

Lab/Branch: Laboratory of Chemistry and Drug Metabolism 
Clinical Pharmacology Branch 

Section: Naval Screening Laboratory, Jacksonville, FL (J. Mitchell and B. Paul). 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 08 Professional: 01 Others: 0.7 



Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Commercial assays for the detection of drugs of abuse in urine change periodically and must be 
reevaluated for validity of detection. Studies are designed to test the validity of new assays on clinical 
specimens obtained from drug users under controlled conditions. 

Healthy male volunteers with a history of chemical substance abuse participate in these studies. 
Informed consent is obtained and all procedures are approved by the hospital Institutional Review Board. 
Commercial assays for detection of drugs of abuse in urine are tested for validity with specimens 
collected under controlled dosing conditions. A variety of drugs of abuse are studied at various dose 
levels. The results of the assays are compared to GC/MS analyses. 

These studies test the validity of commercial assays on clinical samples instead of "spiked" samples and 
provide unique and valuable information to the military and industry concerning their time course of 
detection, specificity and accuracy. 



166 



Publications: 

Cone, E.J., Yousefnejad, D. and Dickerson, S.L., Validity testing of commercial urine cocaine metabolite 
assays. IV. Evaluation of the EmitR d.a.u.TM Cocaine Metabolite Assay in a quantitative mode for 
detection of cocaine metabolite. J. Forensic Sci. . 35: 786-791, 1990. 

Cone, E.J., Dickerson, S. Forensic Drug Testing for Opiates: IV. Analytical Sensitivity, Specificity and 
Accuracy of Commercial Urine Opiates Immunoassays. L Anal. Toxicol ., in press. 1991. 

Cone, E.J., Darwin, W.D., Dickerson, S. Evaluation of the Abuscreen ONTRAK Assay for Cocaine 
(Metabolite), submitted, Clin. Chem ., 

Cone, E.J. On-site Drug Testing: Expediency Versus Accuracy. Employment Testing , BWR:621-28. 

Abstracts: 

Dickerson. S.L. and Cone, E.J., Drug Assay Development XXII. Evaluation of the AbuscTeen/Ontrak 
Assay for Qualitative Detection of Cocaine Metabolite in Human Urine. American Chemical Society . 
MARM. Madison. NJ. May 23-25. 1990. 

Cone, E.J , Darwin WD. and Dickerson, S.L. Validity Assessment of the Abuscreen ONTRAK Assay for 
Cocaine. Society of Forensic Toxicology . Sept. 11-15,1990. 

Cone, E.J.. Darwin, W.D. and Dickerson. S. Detection of Buprenorphine in Human Biofluids utilyzing 
Diagnostic Products Corporation's (DPC) Double Antibody Radioimmunoassay. The International 
Association of Forensic Toxicologists 27th International Meeting, Perth, Western Australia, Oct. 19-23, 
1990. 

Huestus, M.A.. Cone, E.J., and Mitchell, J. Accuracy of Immunoassays for the Detection of Cocaine 
Metabolites in Urine at current and Proposed NIDA Cutoffs. American Academy of Forensic, Anaheim, 
CA, Feb. 18-23, 1991. 

Cone. E.J. and Mitchell, J. Can the NIDA Cutoffs for Opiate Urine Screening and Confirmation be 
Lowered? American Academy of Forensic, Anaheim, CA, Feb.18-23, 1991. 

Maguire, T., Darwin, W.D.. and Cone, E.J. Drug Assay development XXIII. Simultaneous Assay for 
Cocaine (C), Norcocaine (NCO, Benzoylecgonine (BE) and Ecgonine Methyl Ester (EME) in Human 
Biofluids by GC/MS. American Chemical Society, MARM, Madison, NJ, May 23-25, 1990. 

Darwin, W.D., Maguire, T.E. and Cone.E.J. Simultaneous Assay for Cocaine, Benzoylecgonine. 
Ecgonine Methyl Ester, Norcocaine and Cocaethylene in Human Biofluids. Society of Forensic 
Toxicology, Sept. 11-15, 1990. 



167 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

January 1, 1990 to December 31, 1990 

Title of Project: Detection of Drugs of Abuse in Human Saliva 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief 



Z01 DA 00003-05 CDM 



Others D. Darwin Chemist 

D. Yousefnejad Chemist 
S. Dickerson Lab Tech 
T. Maguire Chemist 
B. Holicky Nurse 



CDM, ARC, NIDA 

ARC, NIDA 
ARC, NIDA 
ARC, NIDA 
ARC, NIDA 
ARC, NIDA 



Lab/Branch: Chemistry and Drug Metabolism Laboratory 
Clinical Pharmacology Branch 

Section: None 



Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 



Total Man Years: 1.4 Professional: 0.1 Others: 



1.3 



Check Appropriate Boxes: 

2£,_ Human Subjects 
_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

The presence of drugs of abuse in saliva of human subjects after drug administration was studied to 
determine the feasibility of drug testing with saliva. 

Healthy male subjects with a history of chemical substance abuse volunteered for the studies. Informed 
consent was obtained and all procedures were approved by the hospital Institutional Review Board. 
Following the administration of cocaine, marijuana or opiate, saliva and blood samples were collected 
periodically. Behavioral and physiological measures were made concurrently with collection of 
biofluids. Samples were analyzed by RIA and gas chromatography/mass spectrometry. Significant 
correlations of blood levels with saliva levels were found for cocaine and opiates. Investigations are 
continuing on marijuana. 

These studies provide the scientific basis for development of new non-invasive tests for drug abuse. 



168 



Publications: 

Darwin, W.D., Maguire, T.. Cone, E.J., Carroll. F.I. Drug Assay Development XXVI. Simultaneous 
assay for cocaine (C), Cocaethylene (CE), Norcocaetbylene (NCE), Benzoylecgonine (BE), Ecgonine 
Methyl Ester (EME) and Norcocaine (NC) In human Biofluids by GC/MS. 25th ACS MARM, Newark, 
DL, May 21-23, 1991. 

Cone, E.J., Dickerson, S.L., Darwin. W.D., Fudala, P. and Johnson. R.E., Elevated Drug Saliva Levels 
Suggest a "Depot-Like" Effect in Subjects Treated with Sublingual Buprenorphine. Comittee on problems 
of Drug Dependence, Richmond, VA June 11-14, 1990. 

Huestis, M.A., Heishman, S.J. and Cone, E.J. Plasma and Saliva THC Levels and Behavioral Effects 
following repeated Marijuana Exposure. Society of Forensic Toxicology, Sept. 1 1-15, 1990. 



169 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00006-04 CDM 

January 1, 1990 to December 31, 1990 

Title of Project: Pharmacokinetics and Pharmacodynamics of Opiate Analgesics 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief CDM, ARC, NJDA 

Others D. Darwin Chemist ARC, NJDA 

D. Yousefnejad Chemist ARC, NIDA 

S. Dickerson Lab Tech ARC, NJDA 

B. Holicky Nurse ARC, NJDA 



Lab/Branch: Laboratory of Chemistry and Drug Metabolism 
Clinical Pharmacology Branch 



Institution and Location: Addiction Research Center, NJDA, Baltimore, MD 21224 

Total Man Years: 1.4 Professional: 0.3 Others: 1.1 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The effects of single doses of intramuscularly administered opiates (heroin, morphine, hydromorphone, 
codeine, oxycodone, oxymorphone) and sublingual buprenorphine are being studied in male human 
volunteers in order to determine the relationship of blood and saliva levels to pharmacologic effects. 
Additionally, the study is being performed to determine if a metabolic marker for heroin abuse can be 
found in urine. 

The subjects are healthy males with a history of heroin abuse. Informed consent is is obtained and all 
procedures are approved by the hospital Institutional Review Board. A total of three test doses (placebo 
and two active doses) are administered in random order. Test measures are made for 24 hrs and 
biological fluids are collected for 7 days after each test. The biological fluids will be analyzed for drug 
and metabolites by chromatographic and immunoassay techniques. 

The significance of this study lies in the potential value of saliva as a new test medium for detection of 
drugs of abuse and the characterization of the time course of excretion of metabolic markers for heroin 
and other opiates in urine and saliva. 



170 



Publications: 

Pickworth, W.B., Welch, P., Henningfield, J.E. and Cone, E.J. Opiate-induced pupillary effects in 
humans, in press, 1990. 

Vaupel, D.B., Cone, E.J., Johnson, R.E. and Su, T-S. Kappa Opioid Partial Agonist Activity of the 
Enkephalin-like Pentapeptide BW942C Based on Urination and in Vitro Studies in Humans and Animals. 
J. Pharmacol. Exp. Ther., 252: 225-34, 1990. 

Furman. W.R., Munster. A.M. and Cone E.J.. Morphine Pharmacokinetics during Anesthesia and Surgery 
in Bum Patients. J. Burn Care Rehabil., 11: 391-4, 1990. 

Cone, E.J., Welch, P., Mitchell, J.M. and Paul, B.D. Forensic Drug Testing for Opiates: I. Detection of 6- 
Acetylmorphine in Urine as an Indicator of Recent Heroin Exposure; Drug and Assay Considerations and 
Detection Times. J. Anal. Toxicol., 15: 1-7 1991. 

Mitchell. J.M.. Paul, B.D., Welch, P. and Cone E.J. Forensic Drug Testing for Opiates; Metabolism and 
Excretion Rates of Morphine in Humans after Morphine Administration. J. Anal. Toxicol., in press. 1991. 

Cone, E.J., Welch, P., Paul, B.D. and Mitchell, J.M. Forensic Drug Testing for Opiates. Urinary 
Excretion rates of Morphine and Codeine following Codeine Administration. J. Anal. Toxicol., in press, 
1991. 

Abstracts: 

Walsh, S.L., Preston K.L., Stitzer, ML., Dickerson, S.L., Cone, E.J., Bigelow.G.E. The Behavioral and 
Pharmacokinetic Profile of High Dose Buprenorphine Administered Sublingually in Humans. Society for 
Neuroscience, New Orleans, LA, Nov. 10-15, 1991. 

Cone, E.J., Welch, P., Paul, B. and Mitchell, J., Specificity of 6-Acetylmorphine in Urine as an Indicator 
of Horoin Use. AAFS, Cincinnati, OH, Feb. 19-24, 1990. 

Mitchell. J., Paul, B., Welch, P. and Cone, E.J., Clinical Studies Indicate that Morphine is not 
Metabolized to Codeine in Human Subjects., AAFS, Cincinnati, OH, Feb., 19-24, 1990. 

Fudala, P.J., Johnson, R.E.. Heishman, S.J., Cone, E.J., and Henningfield, J.E., A Dose Run-up and 
Safety Evaluation of Nalmefene HCI in Human Volunteers. NIDA Research Monograph 95. Rockville: 
The Committee on Problems of Drug Dependence, Inc. 1990: 451-52. 

furman, W.R., Cone. E.J., Bengson, Z.B., Stiff, J.L., and Munster, A.M., Morphine Pharmacokinetics 
During Anesthesia and Surgery in Bum Patients., American Bum Association. Las Vegas, Nevada, 
March 30, 1990. 

Uhl, G.R., Newlin, D.B., Pretorius, M.B., Park, J.S., Darwin, W.D. and Cone, E.J., Antagonist- 
Withdrawal Up-Regulation of Endogenoud Opiate Antinociceptive Systems. Comittee on Problems of 
Drag Dependence, Richmond, VA, June 11-14, 1990. 

Darwin, W.D. and Cone, E.J. Drag Assay Development. XXI. Solid Phase Extration (SPE) of 
Buprenorphine from Human Biofluids. American Chemical Society, MARM, Madison, NJ. May 23-25, 
1990. 

Cone, E.J., Holicky, B., Pickworth, W. and Johnson, R.E. Pharmacologic and Behavioral Effects of High 
Doses of Intravenous Buprenorphine, Committee on Problems of Drag Dependence, Palm Beach, FL, 
June 16-20, 1991. 



171 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00007-03 CDM 

January 1, 1 990 to Decmber 3 1 , 1990 

Title of Project: Pharmacokinetics and Pharmacodynamics of Drugs of Abuse in Hair 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief CDM, ARC, NJDA 

Others B. Holicky Nurse ARC, NJDA 

S. Dickerson Lab Tech ARC, NJDA 

D. Darwin Chemist ARC, NIDA 

D. Yousefnejad Chemist ARC, NIDA 

T. Maguire Chemist ARC, NIDA 

Lab/Branch: Laboratory of Chemistry and Drug Metabolism 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: Addiction Research Center, NJDA, Baltimore, MD 21224 

Total Man Years: 8 Professional: 0.2 Others: 0.6 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Drug residues have been detected in human hair specimens by a variety of analytical techniques. These 
reports have generated substantial interest in using hair as a historical record of drug usage. Studies are 
designed to determine the presence and time course of drugs of abuse in human hair. 

Healthy male volunteers with a history of chemical substance abuse will participate in the study. 
Informed consent will be obtained and all procedures will be approved by the hospital Institutional 
Review Board. Subjects will reside on the clinical ward of the ARC. Head and facial hair specimens 
will be obtained prior to and after administration of drugs of abuse. Blood, saliva and urine specimens 
also will be obtained. Analyses of tissue and biofluids for drug will be performed by radioimmunoassay 
and gas chromatography /mass spectrometry. 

The studies will provide the scientific basis for determining the usefulness of hair as a "historical record" 
for substance abuse. 



172 



Publications: 

Periodicals: 

Cone. E.J. Testing human hair for drugs of abuse. I. Individual dose and time profiles of morphine and 
codeine in plasma, saliva, urine and beard compared to drug-induced effects on pupils and behavior. J. 
Anal. Toxicol. 14: 1-7, 1990. 

Cone. E.J., Yousefnejad, D., Darwin. W.D. and Maguire, T. testing Human Hair for Drugs of Abuse II. 
Identification of Unique Cocaine Metabolites in Hair of Drug Abusers and Evaluation of 
Decontamination Procedures. J. Anal. Toxiol., in press, 1991. 

Chapter: 

Cone, E.J. and Dickerson. S.L. Analysis of human facial hair for morphine and codeine: Excretion 
patterns after single doses. In Proceedings of the International Association of Forensic Toxicolo gists. 
1989, in press. 1990. 

Abstract: 

Maguire, T., Darwin. W.D., and Cone. E.J. Drug Assay Development. XXVII. Simultaneous Assay for 
the detection of Cocaine and Opiates in Human Hair by GC/MS. 25th ACS MARM. Newark, DL. May 
21-23, 1991. 



173 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00008-02 CDM 

January 1. 1990 to December 31, 1990 

Title of Project: Pharmacokinetics and Pharmacodynamics of Methamphetamine 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief CDM, ARC. NIDA 

Others B. Holicky Nurse ARC, NIDA 

D. Darwin Chemist ARC, NIDA 

D. Yousefnejad Chemist ARC, NIDA 

T. Maguire Chemist ARC, NIDA 

Lab/Branch: Laboratory of Chemistry and Drug Metabolism, Clinical Pharmacology Branch 
Section: None 

Institution and Location: Addiction Research Center, NIDA. Baltimore, MD 21224 

Total Man Years: 0.5 Professional: 0.1 Others: 0.4 

Check Appropriate Boxes: 

2£_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

Methamphetamine is an amphetamine-like stimulant with substantial abuse liability by the intravenous 
route. RecenUy, a new form of methamphetamine ("ice") has been reported to be abused via the smoking 
route. 

This study will evaluate various chemical forms of methamphetamine in human volunteer subjects. 
Informed consent will be obtained and all procedures will be approved by the hospital Institutional 
Review Board. Methamphetamine will be administered by the smoking and intravenous routes. The 
pharmacokinetic profile will be determined by analysis of blood samples. The bioavailability and abuse 
liability of the smoked drug will be obtained by comparison to the intravenous route. Urine, saliva, and 
hair specimens will be collected for drug detection studies on methamphetamine. 

These studies will evaluate for the first time, the abuse potential of smoked methamphetamine and will 
allow drug detection methods to be developed for this new form of methamphetamine. 



174 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

January 1. 1990 to December 31, 1990 

Title of Project: Fast Action Dynamics of Marijuana 

Principal Investigators: Cooperating Units 



Z01 DA 00009-02 CDM 



PI 


MA. Huestis 


Fellow 


CDM, ARC, NJDA 


Others 


E.J. Cone 


Chief 


ARC. NJDA 




B. Holicky 


Nurse 


ARC, NIDA 




D. Darwin 


Chemist 


ARC, NJDA 




J. Henningfield 


Chief 


BDL, ARC, NIDA 




A. Sampson 


Nurse 


ARC, NJDA 



Lab/Branch: Laboratory of Chemistry and Drug Metabolism 
Clinical Pharmacology Branch 

Section: None 



Institution and Location: Addiction Research Center, NJDA, Baltimore. MD 21224 

Total Man Years: 0.9 Professional: 0.6 Others: 0.3 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

Although early changes which occur during the smoking of marijuana are more likely to be indicative of 
its mode of action, this phase of the use of marijuana has largely been ignored and very little is known 
regarding what happens to a human subject during the smoking process. 

This study will detail the effects of smoking marijuana cigarettes on a variety of systems including 
physiologic effects, behavior and hormonal systems. In addition, blood and saliva levels will be 
determined during and after smoking. Blood and saliva levels will be compared to drug-induced effects 
and hormonal changes. 

The results from this study will provide the most comprehensive assessment of marijuana's effects that 
occur both during and after smoking and should provide important insight to the mode of action of this 
widely abused drug. 



175 



Publications: 

Heishman, S.J., Huestis, MA.. Henningfield, J.E. and Cone, E.J. Acute and Residual Effects of 
Marijuana: Profiles of Plasma THC Levels, Physiological, Subjective and Performance Measures. 
Pharmacol. Biochem. Beh., 37: 561-65, 1990. 

Huestis, M.A., Heishman, S.J. and Cone, E.J. Profile of Repeated Marijuana Exposure; Plasma and 
Saliva THC Levels, Performance, Physiological and Subjective Effects Following Smoking in a 
Controlled Clinical Setting. The International Association of Forensic Toxicologists 27th International 
Meeting, Perth, Western Australia. Oct. 19-23, 1990. 

Huestis, M.. Sampson, A., Holicky, B. and Cone, E.J. The Fast Action Pharmacodynamics of Marijuana 
Smoking, Committee on Problems of Drug Dependence, Palm Beach, FL, June 16-20, 1991. 



176 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00010-02 CDM 

January 1. 1990 to December 31, 1990 

Title of Project: Methodological Assessment of the Risk of Passive Inhalation of Drugs of Abuse 

Principal Investigators: Cooperating Units 

PI E.J. Cone Chief CDM, ARC. NIDA 

Others D. Yousefhejad Chemist ARC, NIDA 

T. Maguire Chemist ARC, NEDA 

Lab/Branch: Laboratory of Chemistry and Drug Metabolism, Clinical Pharmacology Branch 
Section: None 

Institution and Location: Addiction Research Center, NDDA. Baltimore, MD 21224 

Total Man Years: 0.7 Professional: 0.1 Others: 0.6 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

When drugs of abuse are smoked, volatile components and pyrolysis material escape into the atmosphere. 
Depending on the local environment, bystanders may be exposed to the drug by passive inhalation of the 
contaminated air. 

Present studies are underway to develop means of heating drugs of abuse in a controlled environment and 
measuring air levels of drug in order to evaluate this potential hazard. Initially, free-base cocaine "crack" 
and methamphetamine "ice" will be evaluated for potential passive inhalation exposure. 

Unknowing drug exposure could be dangerous to unsuspecting bystanders, particularly to small children. 
These studies will establish limits of exposure to volatile components of drugs under controlled 
conditions. 



177 



Publications: 

Yousefnejad. D. and Cone, E.J. Drug assay development. XXIV. Determination of cocaine in air by 
capillary gas chromatography/mass spectrometry. American Chemical Society Meeting, 24th MARM. 
May 23-25, 1990. 

Yousefnejad, D., Cone, E.J. Drug Assay Development. XXV. Determination of Cocaine in Room Air By 
GC/MS. 25th ACS MARM. Newark, DL, May 21-23. 1991. 

Yousefnejad, D. and Cone, E.J. Drug Assay Development XXTV. Determination of Cocaine in Air by 
Capillary Gas Chromatography/Mass Spectrometry. American Chemical Society, MARM, Madison. NJ. 
May 23-25, 1990. 



178 



3. Neuroendocrinology/Immunology Laboratory - Jack E. Henningfield, Ph.D., Acting Chief 

The Neuroendocrinology /Immunology laboratory investigates mechanisms by which substances of abuse, 
particularly cocaine and tetrahydrocannabinol, act to alter two of the body's homeostatic mechanisms; the 
endocrine and immune systems. This is important because neurosecretion is a marker of CNS function 
and may be altered markedly by substance abuse. Further, neurohormones interact with the immune 
system and may be important regulators of immune function. Examination of alterations in endocrine 
function simultaneously with immune function may help to explain the relative contributions of 
neuroendocrine and local regulation to alter immune function in substance abusers, and also mechanisms 
by which substances of abuse alter immune function. The more rapid progress to AIDS and death in 
substance abusers with HIV infection when compared with other groups of HIV infected individuals may, 
thus, be explained. 

Neuroendocrine and immune function in substance abusers on the ARC research ward are being 
examined, thereby providing relevant clinical findings and developing animal models that mimic the 
clinical findings. Neurohormones are monitored as CNS markers in studying the physiological and 
pharmacological effects of substances of abuse on the hypothalamo-pituitary system in clinical and basic 
research settings. The neuroendocrine hormones are regulated by releasing factors and neurotransmitters 
released from the hypothalamus, steroid hormones and other feedback loops. Each of these may be 
perturbed by substances of abuse. Consideration of perturbators or regulators and subsequent 
measurement of the release neurohormones may be used to suggest mechanisms of drug actions on the 
hypothalamo-pituitary system. In animal models, the relevance of the hypothalamo-pituitary responses to 
CNS mechanisms is being explored. 

The goals of the laboratory are: 

a. to investigate the disturbances of neuroendocrine secretion caused by substances of abuse. 

b. to continue research into basic mechanisms of neuroendocrine secretion, endocrine-mediated 
immune function and their relationship to substance use or abuse. 

c. to investigate the interactions between substances of abuse, neurohormones and altered immune 
function, and to address the hypothesis that substances of abuse act as immunodepressors and may be 
cofactors in the development of AIDS in HIV infected substance users. 

d. to determine the HIV antibody status of ARC volunteers, research subjects and addicts in the 
NFDA HTV-antibody prevalence study. 

e. to provide quantitation of drug concentrations in body fluids and tissue extracts. 

Techniques employed by the NEI Laboratory 

a. radioimmunoassay of hormones and substances of abuse in human and rat body fluids and 
tissues. 

b. quantitation of hormones and neurotransmitters by HPLC. 

c. clinical tests of neuroendocrine secretion. 

d. in vivo and in vitro experimental techniques such as indwelling catheterization and portal vein 
cannulation; perfusion, reverse hemolytic plaque assay, in vivo microdialysis, lymphocyte 
function tests and receptor quantitation for studying neuroendocrine and immune function in 
animals. 

Neurosecretion in Substance Abuse 

Release of hormones from the hypothalamo-pituitary axis is a marker of CNS function. CNS function is 
being examined by studying neuroendocrine secretion particularly in volunteers who have used 
substances of abuse (cocaine and cannabinoids) over prolonged periods of time. Cocaine acts primarily 
through dopaminergic mechanisms and, used chronically, may result in either dopamine depletion or 



179 



hypersensitivity to dopamine-mediated responses. By examining neuroendocrine parameters in detail, 
the relative contribution of dopamine and other neuroendocrine regulators in the hypothalamo-pituitary 
axis may be elucidated. In volunteers who used cocaine heavily and who were abruptly withdrawn from 
the drug on the ARC ward, prolactin concentrations throughout the day and over 3 weeks following 
withdrawal were elevated compared with volunteers who did not use cocaine. (Dopamine is a tonic 
inhibitor of prolactin release. ) Resting levels in hormones of the hypothalamo-pituitary-adrenal axis were 
not altered, suggesting chronic cocaine use has a greater and long lasting effect on dopaminergic 
hypothalamic neurons. In male volunteers who had a heavy use of cannabinoids, prolactin secretion was 
decreased compared with secretion in occasional users. 

These detailed studies of endocrinological alterations at rest are being extended by examining 
neurosecretion under stimulated conditions to obtain further information on the relative contributions of 
particular neurotransmitters to the control of neuroendocrine secretion in chronic substance abuse. 
Evidence suggests that hypersensitivity of responses in both the dopaminergic and serotonergic systems 
occurs. This is being explored in detail. The gradual loss of the hypersensitivity created by substance 
use and observed during withdrawal may be important in relapse. 

Using the rat, a model for chronic cocaine use has been established, passively administering smaller 
doses of cocaine than are usually used in such paradigms, and in multiple divided doses over 10 days. 
This regimen resulted in endocrinological findings similar to those observed in the male volunteers 
withdrawing from cocaine. Prolactin levels were measured prior to and following cocaine administration 
immediately following cocaine administration. Immediately following cocaine administration in the rats, 
prolactin was increasingly depressed until on day 10 it was below detectable levels but increased to 
hyperprolactinemic levels 24 hours after the last injection. To examine the relevance of alterations 
observed in the hypothalamo-pituitary system, the brains of these animals are being examined to establish 
whether alterations in neurotransmitters the tuberoinfundibular neurons reflect those in other brain 
regions. By reverse hemolytic plaque assay it has been established that cocaine has no direct action on 
pituicytes, but release of prolactin from individual cells in response to dopamine is greater and occurs 
from more cells in the cocaine treatment rats suggesting hypersensitivity to dopamine. Future studies 
will concentrate on comparing CNS mechanisms with neurosecretory mechanisms in this model in 
response to various stimulators and depressors administered in vivo and on examination of the anatomy 
and mechanisms of alteration and their response to therapeutic agents and the study of withdrawal 
phenomena. The rat model will also enable examination of the effects of cocaine on other systems, 
particularly cardiac and immunological systems. 

Although dopaminergic mechanisms may be most important in cocaine abuse, cocaine also affects 
serotonergic mechanisms. The serotonergic agonist, metachlorophenylpiperazine (mCPP) has been 
shown to release prolactin and Cortisol which may be followed as in vivo markers of its CNS action. 
Studies of the acute effects of mCPP compared with those of fenfluramine, which has been more 
commonly studied, are complete. Studies examining the more chronic effects of mCPP are being carried 
out in humans and rats. Provocative tests suggest that serotonergic mechanisms in chronic cocaine users 
may be hypersensitive, in which case a chronic serotonergic agonist such as mCPP may be important in 
therapy. Therefore, in the future, the interactions of mCPP in cocaine treated animals or in humans 
withdrawing from cocaine are to be examined. 

Mechanisms of Endocrine-Mediated Immune Function 

Prolactin is an important immune-regulator. Because marked disturbances of prolactin secretion in 
substance users have been observed and a rat model for cocaine has demonstrated similar alterations in 
prolactin release, studies have commenced to examine the mechanisms of alterations of immunological 
function by substances of abuse. Growth hormone may have similar importance in immune-regulation 
and is similarly disturbed in cocaine abusers. The role of growth hormone in lymphocyte function will be 
investigated in new studies. These studies involve examining prolactin-sensitive ornithine decarboxylase 



180 



activity and other parameters of lymphocyte activity in animals chronically administered substances of 
abuse. The immunosuppressant properties of substances of abuse will be assessed in rats by observing 
the rat, and measuring neuroimmunomodulating parameters (including prolactin and growth hormone) 
and lymphocyte function. 

Interactions Between Substance Abuse, the Endocrine and Immune System 

The immunologic effects of substances of abuse have been and are being assessed on a clinical level in 
collaboration with Dr. William Adler's immunology group at the Gerontology Research Center. Amyl 
nitrite has been implicated as an etiological factor in the development of Kaposi's Sarcoma, the most 
common malignant complication of AIDS in homosexuals. In a study where amyl nitrite was 
administered to volunteers it was found that it depressed natural killer cell function during administration 
and that a rebound occurred in other parameters. This suggested an alternating, relative 
immunodepression/immunostimulation cycle could be induced by using the drug which, in turn, could 
promote opportunistic infections or compromise immuno-surveillance. Similar immunological 
techniques have been used to examine endocrine and immune function in tetrahydocannabinol 
administration. Lastly, immune function during cocaine and nicotine administration is being assessed. 
These studies are unusual because they are conducted in a controlled environment and subjects are used 
as their own controls. At this stage, studies are carried out in HTV seronegative volunteers. Future 
studies will be made with HTV seropositive volunteers. 



181 



Publications 

Ulrichsen, J.. Partilla, J.S., and Dax. E.M. Long-term administration of m-chlorophenyl-piperazine 
(mCPP) to rats induces changes in serotonin receptor binding, dopamine levels and locomotor activity 
without altering prolactin and cortiscosterone secretion. Psvchopharmacologv . Submitted. 

Buckenmeyer, P.J., Goldfarb, A.H., Partilla, J.S., Pineyro, M.A., and Dax, E.M. (1990) Endurance 
training, not acute exercise, differentially alters beta-receptors and cyclase in skeletal fiber types. Am 
Physjol 258(1 Pt 1): E7I-7. 

Dax. E.M. (1990) Drug dependence in the differential diagnosis of allergic respiratory disease. Annal 
Allergy 64 (3): 261-3. 

Dax, E.M., Adler, W.H., Nagel, J.E., Lange, W.R. and Jaffe, J.H. inhalation of volatile nitrites induces 
changes in in vitro immune function. Clin, and Exp. Immunol. Submitted. 

Dax, EM., Partilla, J.S., Pineyro, M.A., and Gregerman, R.I. (1990) Altered glucagon- and 
catecholamine hormone-sensitive adenylyl cyclase responsiveness in rat liver membranes induced by 
manipulation of dietary fatty acid intake. Endocrinology 127 (5): 2236-2240. 

Dax, EM., Pilotte, N.S., Adler, W.H., Nagel, J.E., and Lange, W.R. (1989) The effects of 9-ene- 
tetrahydrocannabinol on hormone release and immune function. J Steroid Biochem . 34 (1-6): 263-270. 

Dax, E.M., Pilotte, N.S.. Adler, W.H., Nagel, J.E. and Lange, W.R. (1990) Short-term 

w"tetrahydrocannabinol (THC) does not affect neuroendocrine or immune parameters. NIDA 
Monograph . In Press. 

Fudala, P.J., Jaffe, J.H, Dax, EM., Johnson, R.E. ( 1990) Use of buprenorphine in the treatment of opioid 
addiction II: Physiological and behavioral effects of daily and alternate day administration and abrupt 
withdrawal. Clin Pharmacol Ther 47 (4): 525-34. 

Lange, W.R., Ball. J.C.. Dax, E.M. et al. A follow-up study of a parent HIV seropositivity among 
parenteral drug abusers in 1971-72. In press. 

Lange, W.R., Dax, EM., Kovacs, R., Kreider, S., and Frame, J. (1989) Are missionaries at risk of 
AIDS? Southern Medical Journal 82. 1075-1078. 

Lange, W.R., Fudala, P.J., Dax, E.M., Johnson, R.E. (1990) Safety and side-effects of buprenorphine in 
the clinical management of heroin addiction. Drug Alcohol Depend 26 ( 1 ): 19-28. 

Litow, R., Robinson. N., Herning, R., Jaffe, J.H. and Dax, E.M. Cognitive function and EEG changes 
with the inhalation of amyl-nitrite. Psvchopharmacologv . Submitted. 

Pilotte, N.S., Mitchell, W.M., Sharpe, L.G., de Souza, E.B. and Dax, E.M. Chronic cocaine 
administration and withdrawal from cocaine modify central neurotensin receptors in rats. SYNAPSE . In 
Press. 

Pilotte, N.S., Sharpe, L.G., and Dax, E.M. (1990) Multiple, but not acute, infusions of cocaine alter the 
release of prolactin in male rats. Brain Res 512 : 107-112. 

Ulrichsen. J., Partilla, J.S., and Dax. E.M. Long-term administration m-chlorophenyl-piperazine (mCPP) 
to rats induces changes in serotonin receptor binding, dopamine levels and locomotor activity without 
altering prolactin and cortiscosterone secretion. Psvchopharmacologv . Submitted. 



182 



Weddington. W.W.. Brown. B.S., (1990) Haertzen, C.A., Cone, E.J., Dax, E.M.. Heming, R.I., 
Michaelson, B.S. Changes in mood, craving, and sleep reported by male cocaine addicts during acute 
abstinence: A controlled, residential study. Arch Gen Psvch 47 (9): 861-868. 

ABSTRACTS 

Pilotte. N.S., Johnson. R.L.. and Dax, E.M. ( 1990) Chronic cocaine in vivo modifies prolactin release 
after dopamine in vitro. Presented at 2nd International Congress of Neuroendocrinology, Bordeaux, 
France. Abst. #P4.93. 

Pilotte. N.S.. Mitchell, W.M., Sharpe, L.G.. de Souza, E.B., and Dax, E.M. (1990) Cocaine-induced 
reduction in neurotensin binding in midbrain is reversed during withdrawal from cocaine. Committee on 
Problems of Drug Dependence, Richmond, Virginia. 

Newlin, D.B., Pretorius, M.B., Wong, C. and Dax, E.M. (1990) Acute marijuana smoking reduces vagal 
tone. Committee on Problems of Drug Dependence, Richmond, Virginia. 

Fralich. J., Lange, R.W., and Dax, E.M. (1990) Prevalence trends of HIV infection among drug abusers 
in Baltimore. Maryland. Sixth International Conference on Aids, San Francisco, California. 

Dax, E.M. and Pilotte, N.S. (1990) Growth hormone (GH) release is altered in men who abruptly cease 
long-term cocaine. Presented at 2nd International Congress of Neuroendocrinology, Bordeaux, France. 
Abst. #P3.59. 

Pilotte, N.S., Sharpe, L.G. and Dax, E.M. (1990) Chronic cocaine modifies growth hormone release after 
5-hydroxytryptophan in male rats. Presented at 20thAnn. Mtg. Soc. Neuroscience, St. Louis, Mo. Abst. 
#291.7. 

Sharpe, L.G., Pilotte, N.S., Mitchell, W.M., de Souza, E.B., and Dax, E.M. (1990) Withdrawal from 
chronic cocaine decreased dopamine transporter sites in the rat nucleus accumbens (NAC). Presented at 
20th Ann. Mtg. Soc. Neuroscience, St. Louis, Mo., Abst. #111.18. 



183 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

January 1, 1990 to December 31, 1990 



Z01 DA 00004-04 NEI 



Title of Project: Inhalable Nitrites - Immune Function and Abuse Potential 
Principal Investigators: 



PI: 


E.M. Dax 


Laboratory Chief 


NEI.ARCNIDA 


Others: 


J.H. Jaffe 


NIDA 






W.R. Lange 


Medical Director 


ARC. NIDA 




R. Herning 


Laboratory Chief 


CHP, ARC, NIDA 




R.M. Litow 


Research Technologist 


NEI, ARC, NIDA 




N. Robinson 


Registered Nurse 


NEI, ARC, NIDA 




W.H. Adler 


Clin. Immunology Section 


GRC, NIA 




J. A. Nagel 


Clin. Immunology Section 


GRC, NIA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 



Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 



Total Man Years 



Professional: 0.25 Others: 0.75 



Check Appropriate Boxes: 

X_ Human Subjects 
_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

The intake and frequency of inhalation of volatile nitrites has been associated with the incidence of 
Kaposi's sarcoma in people suffering from AIDS. Animal and in vitro lymphocyte studies have shown 
that immune cell function can be altered by these agents. However, no study has related directly the 
effects of nitrites administered in vivo to disturbances of immune function in humans. Thus, a study has 
been conducted in healthy, HIV negative volunteers. An initiation protocol in which the subject inhaled 
3 doses of amyl nitrite for 3 days and 1 dose on the fourth day has been conducted. In an extended 
protocol a second group of volunteers were administered subsequent, single inhalations of nitrite 3-4 days 
apart, to a total of 13 inhalations over 3 weeks. A battery of immune function tests in the subjects' 
lyphocytes, was carried out on 2 occasions prior to the inhalation protocol, immediately following the last 
dose, and at 1,4, and 7 days after the last dose. Results showed a decrease in natural killer cell activity, 
the lymphocyte function reputedly responsible for tumor cell scavenging. The single doses of nitrite 
administered at 3-4 day intervals continued to suppress this activity. Lymphocyte numbers and subsets 
were not altered during the inhalation protocols, but showed a non-specific rise on cessation of the drug. 
Discrepencies between mitogen stimulated [3H]thymidine incorporation, a measure of the activity 
potential of lymphocytes, and antibody production by the T lymphocyte-dependent, B-cells indicated a 
deficit in T-cell function during nitrite exposure. 

The nitrites were demonstrated to have minimal abuse potential. 



184 



Publications 



Litow, R.. Robinson. N., Herning, R., Jaffe, J.H. and Dax, E.M. Cognitive function and EEG changes 
with the inhalation of amyl-ni trite. Psychopharmacology. Submitted. 

Dax, EM.. Adler. W.H, Nagel. J.E., Lange, W.R. and Jaffe, J.H. Inhalation of volatile nitrites induces 
changes in in vitro immune function. Clin, and Exp. Immunol. Submitted. 



185 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00005-04 NEI 

January 1. 1990 to December 31, 1990 

Title of Project: HIV Prevalence: In Depth Survey of Baltimore 

Principal Investigators: 

PI: E.M. Dax Laboratory Chief NEI, ARC, NIDA 

Others: W.R. Lange Medical Director ARC, NIDA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 
Total Man Years: 10 Professional: 0.25 Others: 0.75 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

The seroprevalence of HIV antibodies in surveyed intravenous substance users (IVSUs) who were either 
recently enrolled into treatment or were on a waiting list for enrollment was 29%. The rate among ARC 
research subjects with parenteral drug use histories has averaged 24%, and among area prostitutes with 
heavy drug use histories, 34%. In Baltimore, 94% of IVSUs had shared needles, and even though HIV 
seropositivity was not assoicated with a needle-sharing history, there was an association between the 
intensity of sharing and the probability of being seropositive. A much stronger association was observed 
between seropositivity and "shooting gallery" visitation, suggesting that this milieu of sharing, rather than 
other environments, is the real risk factor. 

Very distinct ethnic group differences in HIV infection were observed, with Blacks being much more 
likely to be seropositive than Whites (odds ratio = 8.18, 95% CI 3.35-19.97). There was no significant 
difference in HrV infection between Blacks in Baltimore and in New York City. Shooting gallery 
visitation appears to be much more a phenomenon among Black IVSUs than it is in White (X2 = 8.23, 
p<0.01). HIV infection has appreciably penetrated Baltimore's addict community. The overall 
seroprevalence rate in Baltimore in 1986 (29%) approximated that of New York in 1979 (27%) where the 
rate subsequenUy jumped to 58% in some areas by 1984 and has increased to 60% in 1987. 

Hepatitis antigen and antibody status of these subjects has been assessed There was no concordance of 
Hepatitis B infection and HIV infection. Other data concerning Hetatitis D is being analysed. 

The second wave of this study is being initiated. 



186 



NOTICE OF INTRAMURAL RESEARCH PROJECT 



Z01 DA 00006-04 NEI 



January 1. 1990 to December 31, 1990 

Title of Project: Cannabinoids and Their Effects on the Immune System and Cognitive Function 

Principal Investigators: 



PI: 


EM. Dax 


Laboratory Chief 


NEI, ARC, NTDA 


Others: 


W.R. Lange 


Medical Director 


ARC, NIDA 




N.S. Pilotte 


Staff Fellow 


NEI, ARC, NIDA 




R.M. Litow 


Research Technologist 


NEI, ARC, NIDA 




J.S. Partilla 


Chemist 


NEI, ARC, NIDA 




N. Robinson 


Registered Nurse 


NEI, ARC, NIDA 




J.R. Mahaffy 


Registered Nurse 


NEI, ARC, NIDA 




W.H. Adler 


Clin. Immunology Section 


GRC, NIA 




J. A. Nagel 


Clin. Immunology Section 


GRCNIA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 
Total Man Years: 2 Professional: 0.5 Others: 1.5 



Check Appropriate Boxes: 

X_ Human Subjects 
_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

Delta-9-tetrahydrocannabinol (THC) has been hypothesized to influence immune function. However, 
this has not been investigated in a comprehensive fashion in humans. The purpose of this study is to 
measure and study the effects of THC on immune function. To investigate immune-endocrine 
correlations, hormone parameters defining the activity of the hypothalamo-pituitary-adrenal axis have 
been measured during THC administration. (The effects of THC on cognitive function will also be 
investigated. ) Experienced THC users have been recruited for study. Immune function of lymphocytes 
in vitro has been investigated during orally administered and inhaled THC and during the washout phase. 



187 



Publications 



Dax, EM., Pilotte, N.S., Adler, W.H., Nagel, J.E., Lange. W.R. (1990) The effects of 9-ene- 
tetrahydrocannabinol on hormone release and immune function. J. Steroid Biochem, 34: 263-270. 

Dax, EM., Pilotte, N.S., Adler, W.H., Nagel, J.E. and Lange, W.R. (1990) Short-term 

W°tetrahydrocannabinol (THC) does not affect neuroendocrine or immune parameters. Committee on 
Problems of Drug Dependence, Richmond, Virginia. 

Newlin, D.B., Pretorius, M.B., Wong, C. and Dax, E.M. (1990) Acute marijuana smoking reduces vagal 
lone. Committee on Problems of Drug Dependence, Richmond, Virginia. 

Robinson, N., Lange, W.R., Dax, E.M. EKG alterations following W"tetrahydrocannabinol. 
(Submitted). 



188 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00007-04 NEI 

January 1. 1990 to December 31, 1990 

Title of Project: Neuroendocrine Secretion During Cocaine Withdrawal 

Principal Investigators: 

PI: Elizabeth Dax. M.D. Ph.D. Laboratory Chief NEI, ARC, NIDA 

Others: W. Weddington. M.D. TEI NIDA 

Nancy Pilotte, Ph.D. Staff Fellow NEI, ARC, NIDA 

Edrich Anderson. R.N. Registered Nurse NEI, ARC, NTDA 

Teri Gendron Research Technologist NEI, ARC, NIDA 

Lab/Branch: Neuroendocrinology/Immunology Branch 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3 Professional: 1 Others: 2 

Check Appropriate Boxes: 

X_ Human Subjects Human Tissues _ Neither 

_ Minors 
Interviews 



Summary Work 

Cocaine withdrawal (and withdrawal from other drugs) is associated with CNS disturbances which are 
reflected in altered neurohormonal secretion, secondary to CNS neurotransmitter alterations. In men 
known to be cocaine abusers, the secretion of neurohormones has been examined during cocaine 
withdrawal. Prolactin secretion is under tonic inhibition by dopamine from the hypothalamus. The 
neurotransmitter most closely associated with Cortisol release is serotonin. In the volunteers withdrawn 
from cocaine, prolactin levels were higher than in men who had not ever taken cocaine, and the diurnal 
rhythms in prolactin secretion were disturbed. The men were followed for up to 21 days with little 
change in the profiles of prolactin release. Cortisol levels and rhythms were similar to controls over this 
withdrawal period. These results suggest that chronic cocaine abuse results in dysfunction of dopamine 
mediated mechanisms of neurosecretion. 

The alterations in the hypothalamo-pituitary-adrenal axis resulting from chronic cocaine abuse, is being 
further defined. This will enable study in volunteers whose serotonergic and dopaminergic functions are 
predictably manipulated, with tests that perturb the hypothalamic-pituitary-adrenal axis at a known level. 
Standard endocrine diagnostic tests (TRH, CRF stimulation and L-dopa suppression) in conjunction with 
drugs that perturb dopaminergic and serotonergic function have been carried out. The study will provide 
further information on dopaminergic control of hormonal secretion and its role in maintaining diurnal 
rhythms of hormones. It may provide an important means of assessing the efficacy of treatment 
protocols. Mechanisms of these changes is being investigated in a rat model of cocaine withdrawal. 



189 



Publications 



Weddinglon, WW., Brown, B.S., Haertzen, C.A., Cone, E.J., Dax, E.M.. Herning, R.I., Michaelson. B.S. 
Changes in mood, craving, and sleep reported by male cocaine addicts during acute abstinence: A 
controlled, residential study. Arch Gen Psych 47 (9): 861-868. 

Dax, E.M. and Pilotte, N.S. Growth hormone (GH) release is altered in men who abruptly cease long- 
term cocaine. Presented at 2nd International Congress of Neuroendocrinology, Bordeaux, France, June. 
1990. Abst. #P3.59. 



190 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00008-04 NEI 

January 1, 1990 to December 31. 1990 

Title of Project: The Effects of Cocaine on Hormone Secretion from the Anterior Pituitary 

Principal Investigators: 

PI: E.M. Dax Laboratory Chief NEI. ARC, NIDA 

N.S. Pilotte Staff Fellow NEI, ARC, NJDA 

Others: L.G. Sharpe Research Psychologist BVL, ARC, NIDA 

Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 

21224 

Total Man Years: 2 Professional: 1 Others: 1 

Check Appropriate Boxes: 

_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

Although there is a large body of evidence linking cocaine use with central dopamine function, little is 
known about the direct or indirect effects of cocaine on the release of hormones from the anterior 
pituitary. The release of prolactin (PRL) is inhibited by dopamine originating from a discrete population 
of hypothalamic neurons. Thus, prolactin release is an indirect measure of dopamine release. 

We have found that rats receiving daily i.v. infusions of cocaine are mildly hyperprolactinemic before the 
daily session of cocaine as early as 5-10 days after initiation of the treatment. Furthermore, PRL is 
decreased markedly at the end of the infusion period. This persistent effect appears to be related to the 
ability of cocaine to block the uptake of released dopamine and a possible decrease in the ability of the 
neuron to release more dopamine in the absence of cocaine. In contrast, dopamine plays little if any role 
in the normal release of growth hormone (GH) from the anterior pituitary. Thus, cocaine does not affect 
the peripheral concentrations of GH unless the release of this hormone is stimulated by serotonin. The 
ability of serotonin to release GH is diminished immediately after cocaine treatment, perhaps reflecting 
the loss of access to a readily-releasable pool of GH, a functional uncoupling of serotonin to its receptors 
on neurons that secrete growth hormone-releasing factor, or a desynchronization of the normal pulsatile 
circadian rhythm of the release of GH. 



191 



Publications 

Pilotte, N.S., Sharpe, L. G. and Dax. E.M. (1990) Multiple, but not acute, infusions of cocaine alter the 
release of prolactin in male rats. Brain Res. 512 : 107-1 12. 

Dax, E.M. and Pilotte, N.S. Growth hormone (GH) release is altered in men who abruptly cease long- 
term cocaine. Presented at 2nd International Congress of Neuroendocrinology, Bordeaux, France, June, 
1990, Abst. #P3.59. 

Pilotte. N.S.. Sharpe, L.G. and Dax, E.M. Chronic cocaine modifies growth hormone release after 5- 
hydroxytryptophen in male rats. Presented at 20th Ann. Mtg. Soc. Neuroscience, St. Louis, Mo. Abst. 
#291.7 



192 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00010-03 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: Development of Monoclonal Antibodies to Drugs and Hormones 

Principal Investigators: 

PI: E.M. Dax Laboratory Chief NEI,ARC,NJX>A 

Others: C. Dersh Chemist NEI, ARC, NEDA 

R. Zaczek Staff Fellow ARC, NIDA 

Lab/Branch: Neuroendocrinology/Immunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2 Professional: .25 Others: 1.75 

Check Appropriate Boxes: 

_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

Usually antibodies to treatment drugs, substances of abuse or hormones are not commercially available 
unless there is wide market. Treatment drugs, such as buprenorphine, are unlikely to have antibodies 
developed until the efficacy of the drug is established. Therefore, this laboratory is developing antibodies 
to buprenorphine in order to establish a radioimmunoassay. Mice have been immunized with 
buprenorphine and demonstrate the presence of antibodies on initial tests. Spleen cells will be harvested 
and fused to myeloma cells to produce cell lines. Testing for production of antibodies in the cell line will 
be carried out and any clones producing highly specific and high affinity antibodies will be isolated. 
Buprenorphine antibodies will be used to establish a radioimmunoassay so that the drug may be 
quantitated in urine, plasma and tissue extracts. Other drugs that antibodies may be raised against 
include amphetamine and metamphetamine, cocaine and cocaine metabolites, and 
metachloropheylpiperazine (mCPP). Antibodies are also presenUy being raised against vasopressin and 
rat prolactin, for which commercially available antibodies are limited. 



193 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00011-03 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: Neuroendocrine Correlates of HIV Infection and the Development of ARC and AIDS 

Principal Investigators: 

PI: E.M. Dax Laboratory Chief NEI, ARC, NEDA 

W.R. Lange Medical Director ARC, NIDA 



Others: Lawrence Brown, M.D., M.P.H., Vice President for Research and Medical Affairs, 
Addiction Research and Treatment Inc., Brooklyn, N. Y. 

Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 
Total Man Years: 1 Professional: 0.5 Others: 0.5 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

The human immunodeficiency virus infects the central nervous system resulting in a wide range of 
neurological deficits. One problem in management of HIV-infected people is predicting the disease's 
prognosis and course. The control of the neuroendocrine system and many feed back loops both 
endocrine and immunological are a property of the CNS, particularly of the hypothalamus. Several 
studies have shown disruption of neuroendocrine function. In a large group of drug abusers the 
neuroendocrine/endocrine status will be correlated with HIV status, clinical history, drug history, and 
presence of opportunistic infections. To date, clinical data from 800 patients have been collected. Their 
HIV antibody status and hormonal measurements are being assessed. 



194 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00012-03 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: HTV Sero-status in Missionaries From Africa, 1968-1983 

Principal Investigators: 

PI: W.R. Lange Medical Director ARC, NJDA 

E.M. Dax Laboratory Chief NEI, ARC, NDDA 

Lab/Branch: Neuroendocrinology/lmmunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1 Professional: 0.5 Others: 0.5 

Check Appropriate Boxes: 

Human Subjects X_ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

The origins and timing of HIV spread into the U.S. remain in question. One possibility is that alterations 
in the virus' genetic makeup led to its current pathogenic properties, but a similar virus may have been 
transported from Africa where HTV is endemic. We have screened approximately 6000 plasmas from 
missionaries travelling between Africa and the U.S. between 1968 and 1983. Although the group may be 
considered low risk for sexually transmitted diseases, they are a group with high casual contact with the 
African people. Approximately 200 plasmas were found to be positive on ELISA screening but none was 
found to have HIV specific proteins detected by Western Blot. Further analysis of the HTV proteins are 
being carried out. Selected plasmas are being screened for related virus, including HTLV1. 



195 



Publications 

Lange, W.R.. Dax. E.M., Kovacs. R.. Kreider. S., and Frame, J. (1989) Are missionaries at risk of 
AIDS? Southern Medical Journal 82: 1075-1078. 



196 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00013-03 NEI 

January 1. 1990 to December 31, 1990 

Title of Project: Neuroendocrine Correlates of Aggressive/Impulsive Behavior 

Principal Investigators: 

PI: E.M. Dax Laboratory Chief NEI, ARC, NJDA 

Others: C.S. Contoreggi Staff Fellow NEI, ARC. NJDA 

D. Fishbein Guest Scientist ARC, NIDA 

J.H. Jaffe NIDA 



Lab/Branch: Neuroendocrinology /Immunology Branch 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 21224 
Total Man Years: 4.0 Professional: 1.5 Others: 2.5 

Check Appropriate Boxes: 

X_ Human Subjects _ Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

When men grouped according to their aggressive/impulsive scores on standard psychological tests, are 
challenged with a serotonergic stimulator, such as fenfluramine, the neuroendocrine response is 
attenuated in the more aggressive, more impulsive men, suggestive of alterations in central serotonergic 
mechanisms. Further, results suggest that hostility ratings decrease with fenfluramine administration 
suggesting possible treatment rationales for the study. Aggression and impusivity may be important 
personality characteristics in initiating and perpetuating addictive behavior. In order to investigate 
mechanisms of this behavior, establish neuroendocrine markers, suggest treatment possibilities and assess 
the efficacy of treatment paradigms, we are extending these studies with the more specific serotonergic 
agonist, metachlorophenylpiperazine (mCPP). We are examining whether mCPP administration gives 
similar results to fenfluramine. Subsequent studies will examine serotonergic as well as dopaminergic 
secretion in greater detail. Using neurohormones as markers of these responses, secretion will be 
examined in the presence of either a serotonergic or dopaminergic agonist (mCPP or bromocryptine, 
respectively). Neuroendocrine provocation tests will be used to further define alterations of function in 
the aggressive men. 



197 



Publications 



Contoreggi, C.S.. Fishbein, D.H., Jaffe, J.H. and Dax, E.M. Comparison of neuroendocrine effects of 
fenfluramine and metachlorophenylpiperazine in male volunteers. In preparation. 



198 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00014-02 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: The effects of cocaine on prolactin secretion from single cells of the anterior pituitary 
gland 

Principal Investigators: 

PI: N.S. Pilotte Staff Fellow NEI, ARC, NIDA 

Others: EM. Dax Laboratory Chief NEI, ARC, NIDA 

R.L. Johnson Biologist NEI, ARC, NIDA 

Lab/Branch: Neuroendocrinology/Immunology Laboratory 

Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1 Professional: .5 Others: .5 

Check Appropriate Boxes: 

Human Subjects Human Tissues _ Neither 

_ Minors 
Interviews 



Summary of Work 

In a recently completed study, we found that in rats that received programmed infusions of 1 mg/kg 
cocaine every 12 min for 2 hr over 10 days, the pre-infusion concentrations of prolactin (PRL) increased 
in a time-dependent manner whereas post-infusion levels of PRL were decreased by cocaine. Because 
dopamine (DA) and PRL are reciprocally related in male rats, these changes could involve modification 
of adenohypophysial dopamine D2 receptors. Dispersed anterior pituitary cells were obtained from rats 
treated chronically with cocaine or saline. The cells were used in a reverse hemolytic plaque assay that 
permitted direct visualization of PRL release from single lactotropes after different treatments in vitro . 
The cells were incubated with media, cocaine, thyrotropin releasing hormone (TRH), or dopamine (DA) 
in vitro . There were 4 major findings. 1) Basal PRL release was greater in rats treated with cocaine: 
more cells secreted PRL and the individual cells secreted more PRL. 2) Cocaine in vitro did not affect 
PRL release. 3) TRH stimulated PRL release similarly from lactotropes of cocaine- or saline-treated 
rats. 4) DA in vitro inhibited PRL release dose-dependently from both cocaine- and saline-treated rats 
when the concentration of DA met or exceeded that observed in hpothalamo-hypophysial portal blood. 
However, lactotropes from cocaine-treated rats were more sensitive to the inhibition by DA. 
Paradoxically, very low concentrations of DA (<10-9M) enhanced PRL release from cells from cocaine- 
treated rats. These data confirm the findings of others that DA-deprived lactotropes release more PRL 
when challenged with low concentrations of DA and suggest that one consequence of chronic use of 
cocaine is a diminished release of DA in the absence of cocaine. 



199 



Publications 

Pilotte. NS, Johnson. RL. Dax. EM. Chronic cocaine in vivo modifies prolactin release after dopamine in 
vitro . Presented at 2nd International Congress of Neuroendocrinology, Bordeaux, France, June 24-29, 
1990. Abst. #P4.93. 



200 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00015-02 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: The effects of cocaine on dopamine release from hypothalamic neurons. 



Principal Investigators: 

PI: N.S. Pilotte 



Staff Fellow 



NEI, ARC, NfDA 



Others: 



L.G. Sharpe 
I.M. Mefford 
E.M. Dax 



Research Psychologist BVL, ARC, NIDA 
Special Expert 
Laboratory Chief 



CP.NIMH 

NEI, ARC, NfDA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1 Professisonal: .75 Others: .25 



Check Appropriate Boxes: 

Human Subjects 

_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

In a recently completed study, we found that in rats that received programmed infusions of 1 mg/kg 
cocaine every 12 min for 2 hr over 10 days, the pre-infusion concentrations of prolactin (PRL) increased 
in a time-dependent manner whereas post-infusion levels of PRL were decreased by cocaine. Because 
dopamine (DA) and PRL are reciprocally related in male rats, these changes could involve modification 
of the release of DA from hypothalamic tuberoinfundibular neurons. We are testing this hypothesis in 
rats treated as described above for 9 days with cocaine or saline. On the 10th day, the hypothalamo- 
hypophysial portal blood will be collected for 30 min before the initiation of passive infusions of cocaine 
or saline, during 60 min of intermittent infusion, and for 30 min following a challenge of amphetamine. 
Arterial blood will be collected concurrendy. These aliquots will be assayed in Dr. Mefford's laboratory 
using microbore high performance liquid chromatography. If there are differences between cocaine- and 
saline-treated animals, another series will be performed with lidocaine as the infusate as a control for the 
local anesthetic effects of cocaine. This experiment will provide the first evidence of cocaine-induced 
modifications of functional DA release coupled to a physiological relevant event, the release of PRL, and 
can serve as a model of the action of cocaine on other central DA systems. 



201 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00016-02 NEI 

January 1, 1990 to December 31, 1990 

Title of Project: Effects of cocaine and withdrawal from cocaine on central receptors for peptides, 
catecholamines, and catecholamine uptake markers. 

Principal Investigators: 

PI: N.S. PUotte Staff Fellow NEI, ARC, NTDA 

Others: W.M. Mitchell Lab Manager NBL, ARC, NIDA 

L.G. Sharpe Research Psychologist BVL, ARC, NIDA 

E.B. de Souza Laboratory Chief NBL, ARC, NIDA 

E.M. Dax Laboratory Chief NEI, ARC, NIDA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3 Professional: 1.5 Others: 1.5 

Check Appropriate Boxes: 

Human Subjects _ Human Tissues _ Neither 

_ Minors 
_ Interviews 

Summary of Work 

Cocaine is thought to produce many of its effects through an interaction with dopaminergic neuronal 
systems. Cocaine's neurochemical effects may include modifications in the numbers of receptors or 
uptake sites for dopamine (DA) or other regulatory peptides colocalized with DA, such as neurotensin 
(NT). If such changes occur, it is not known if they are permanent. Thus, we treated rats with 
programmed infusions of isotonic saline or 1 mg/kg cocaine every 12 min for 2 hr over 10 days and killed 
them within 15 min of the last infusion. Other rats were treated identically, but were killed 10 days later. 
Brains were removed and immediately frozen. Ten micron sections were taken through areas known to 
contain DA perikarya or terminals and binding experiments were conducted on the slices to determine the 
loci and number of binding sites for NT, and desmethylimipramine-insensitive mazindol binding sites to 
mark the DA transporter. Additional sections were taken for analysis of binding of paroxetine, and 
corticotropin releasing hormone. Analysis for NT sites is complete at this time. We found that cocaine 
reduced NT binding sites in the ventral tegmental area, substantia nigra and pars lateralis and that these 
reductions were reversed 10 days later. In addition, NT binding in the prefrontal cortex of rats killed 
within 15 minutes of their final infusion of cocaine was twice that of saline-treated rats and these sites 
were increased by an additional 50% ten days after withdrawal of cocaine. In contrast, the binding of DA 
uptake sites was not changed at the end of the period of cocaine administration, but was reduced in the 
nucleus accumbens ten days after withdrawal of cocaine. The persistent up-and down-regulation of these 
receptors may be intimately involved in the long-lasting behavioral and psychological effects associated 



202 



with the use of cocaine and abstinence from it. 



Publications 



Pilotte, N.S., Mitchell, W.M.. Sharpe, L.G., de Souza. E.B. and Dax. E.M. Chronic cocaine 
administration and withdrawal from cocaine modify central neurotensin receptors in rats. SYNAPSE. In 
Press. 

Sharpe, L.G., Pilotte, N.S., Mitchell, W.M., and de Souza, E.B. Withdrawal of repeated cocaine 
decreases autoradiographic labeling of dopamine transporter in rat nucleus accumbens. Submitted to Eur 
J Pharmacol. 

Pilotte, N.S., Mitchell, W.M., Sharpe, L.G., de Souza. E.B., Dax, E.M. Cocaine-induced reduction in 
neurotensin binding in midbrain is reversed during withdrawal from cocaine. Presented at CPDD, June, 
1990. 

Sharpe, L.G., Pilotte, N.S., Mitchell, W.M.. de Souza, E.B., and Dax, E.M. Withdrawal from chronic 
cocaine decreased dopamine transporter sites in the rat nucleus accumbens (NAC). Presented at 20th 
Ann. Mtg. Soc. Neuroscience, St. Louis. Mo., Abst. #111.18. 



203 



NOTICE OF INTRAMURAL RESEARCH PROJECT 



Z01 DA 00017-02 NEI 



January 1, 1990 to December 31, 1990 

Title of Project: Cardiac effects of I.V. cocaine administration as measured by radionucleotide 
scanning, echocardiography and holter monitoring. 



Principal Investigators: 



PI: 



Others: 



E.M. Dax 
W.R. Lange 
N. Chandra 

C.S. Contoreggi 
J. Fralich 
F. Levin 



Laboratory Chief 
Medical Officer 
Cardiologist 

Assistant Medical Officer 
Physician's Assistant 
Staff Fellow 



NEI, ARC, NJDA 
ARC, NDDA 
FSKMC 

ARC, NJDA 
ARC, NIDA 
ARC, NJDA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2 Professional: 1.5 Others: .5 
Check Appropriate Boxes: 



X_ Human Subjects 
_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

Cocaine use is associated with sudden death which is often due to cardiac complications. The 
mechanism of cocaine's effect on cardiac function is not understood. In healthy volunteers who use 
cocaine, cardiac function will be monitored in the absence and presence of cocaine by holter monitoring, 
echocardiography and radionucleotide (Thallium) scanning. In addition, physiological, 

neuroendocrinological and peripheral nervous system data will be obtained. 



204 



NOTICE OF INTRAMURAL RESEARCH PROJECT 

January 1, 1990 to December 31, 1990 



Z01 DA 00018-02 NEI 



Title of Project: The effect of chronically administered metachlorophenylpiperazine on rat brain 
receptors, neurotransmitters and neuroendocrine hormone secretion. 



Principal Investigators: 

PI: E.M. Dax 



Laboratory Chief 



NEI. ARC. NfDA 



Others: 



J. Ulrichsen 
J.S. Partilla 



Foreign Fellow 
Research Chemist 



NEI, ARC, NIDA 
NEI, ARC, NIDA 



Lab/Branch: Neuroendocrinology/Immunology Laboratory 
Clinical Pharmacology Branch 

Section: None 

Institution and Location: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2 Professional: 1 Others: 1 
Check Appropriate Boxes: 



X_ Human Subjects 
_ Minors 
Interviews 



Human Tissues 



Neither 



Summary of Work 

Metachlorophenylpiperazine (mCPP) is a serotonergic agonist/antagonist which may be useful in 
treatment of disorders such as depression and aggression which are associated with substance abuse. 
mCPP may have therapeutic value for cocaine abuse. Although neuroendocrine and other responses to 
acute treatment with mCPP have been studied, no studies of alterations with chronic administration have 
been made. We will conduct these studies in rats chronically treated with mCPP. Appropriate receptors 
and neurotransmitters along with neuroendocrine responses and behavioral parameters will be 
quantitated. The interaction of mCPP with cocaine effects will be investigated. 

The effects of chronic mCPP on human neuroendocrine responsiveness will also be assessed. Subjects 
will be given the neuroendocrine challenges of TRH- and CRH - stimulation tests after chronic mCPP 
administration. 



205 



Etiology Branch 

David B. Newlin, Ph.D., Acting Chief 

Overview 

The Etiology Branch is concerned with the causes of drug abuse. Research is conducted with human 
subjects in both clinical and laboratory settings. Dr. David Newlin is Acting Chief of the Etiology 
Branch, and a permanent Branch Chief currendy is being recruited 

Vulnerability Laboratory - David. B. Newlin, Ph.D., Acting Chief 

The main focus of the Vulnerability Laboratory concerns mechanisms of addiction, with special emphasis 
on populations who are at high risk for developing drug abuse. Broad secdons of the population are 
exposed to illicit drugs, but only a small percentage develop serious drug problems. The central question 
is what constitutional and environmental factors promote the development of drug abuse, and what are 
the mechanisms by which initial drug use is translated into addicdon. 

High risk populations that are being studied include infants exposed to drugs in utero. individuals at 
genetic or familial risk, individuals at risk due to certain premorbid personality characteristics, 
individuals with genetic markers for alcoholism and/or drug abuse, and established drug abusers who are 
at risk for relapse. 

Several different mechanisms of addiction are being studied that are thought to relate to the process by 
which at-risk individuals develop drug abuse. These mechanisms include attentional deficits in childhood 
and aggressive-impulsive characterisrics in adolescents, electrocortical characteristics, Pavlovian drug 
conditioning mechanisms, enkephalinergic transcriptional processes, deviant responses to common drugs 
that may explain high risk status, individual differences in reward to abused drugs, access to drugs and 
alternative reinforcers. craving for drugs, and withdrawal phenomena. 

Specific studies in the Vulnerability Laboratory span the range from infants to adulthood. We are 
initiating research on infants exposed to cocaine and other drugs in utero which we measure vagal tone, 
response to startle, electroencephalographic activity and developmental course, as well as mother-infant 
interactions. This research is expected to provide invaluable noninvasive physiological and behavioral 
data on outcome in infants exposed to drugs during pregnancy. Simdar research is being conducted with 
8-11 year old children who had been exposed to opiates in utero, compared to chddren whose mothers 
were exposed to opiates after pregnancy (during child-rearing) and mothers not exposed to drugs. This 
research should indicate long-term effects of in utero exposure and child-rearing effects of maternal 
exposure to drugs. 

In the adolescent age range, we are studying responses to alcohol, methylphenidate, and mCPP in young 
men (aged 21-25) who are at high risk for developing drug abuse and alcoholism because of a family 
history of alcoholism or personal history of hyperactivity. This research involves a wide range of 
measures, including autonomic, EEG, neuroendocrine, and subjective report of intoxication and drug 
effects following drug challenge in these individuals. This work in progress is expected to delineate 
differences in acute and chronic drug-induced responses that may account for the increased risk status of 
these individuals. 

Research with gene regulation of the endogenous opiate system has moved into a second phase of 
replication and extension of previous work. We found that chronic treatment with opiate antagonists lead 
to greater anti-nociceptive effects during the antagonist wash-out period. We are currently conducting 
follow-up research to replicate this effect and to determine whether this effect is pre- or post-synaptic. 



206 



Other research on mechanisms of addiction involve studying the acute effects of various abused and 
nonabused drugs on the cardiovascular system in order to elucidate parasympathetic effects of these 
drugs. This research is driven by our linkage hypothesis that proposes central linkage between 
dopaminergic reward mechanisms, psychomotor stimulant effects, and vagally-mediated heart rate 
increases to abused drugs. Following prediction, we have found that alcohol, morphine, cocaine, and 
smoked marijuana decrease vagal tone, in some cases dramatically. This is evidence that the supposed 
sympathomimetic effects of these drugs are largely due to parasympathetic rather than sympathetic 
mechanisms. We are curremly extending this research to replicate the alcohol, opiate, and marijuana 
effects on vagal tone, and to study new drugs such as nicotine, barbiturates, d-amphetamine, 
methylphenidate, mCPP, and naloxone-precipitated withdrawal. 

Our research with induced cocaine craving, in which we found that alcohol intoxication increased self- 
report of cocaine craving, is being extended to follow-up on the large and reliable individual differences 
in cocaine craving. We have found that high cocaine cravers have increased resting vagal tone, THM 
index, and activity, and lower heart rates than low cocaine cravers. This pattern is similar to findings in 
infants in which temperamental traits of "fussiness" have been related to similar cardiovascular and 
behavioral differences. 

Previous research on cognitive and evoked potential correlates of cocaine withdrawal is being extended 
to include longer intervals following cocaine exposure and a more comprehensive cognitive and evoked 
potential battery of measures. This research is providing a very effective documentation of the temporal 
effects of cocaine withdrawal, as well as possible residual effects of cocaine or preexisting characteristics 
of cocaine abusers. 



207 



Publications 

Heming, RE., Glover, B.J., Weddington, WW., Koeppl, B.S., and Jaffe, J.H. Cognitive decrements 
during cocaine abstinence were not related to depression. Biological Psychiatry . Submitted January, 
1990, In revision. 

Weddington, WW., Brown. B.S., Haertzen, C.H., Cone, E.J., Dax, E.M., Herning, R.I., and Michaelson, 
B.S. Changes in mood, craving and sleep during acute abstinence reported by male cocaine addicts: A 
controlled residential study. Archives of General Psychiatry . 47: 861-868, 1990. 

Litow. R.M., Herning, R.I.. Robinson, N., Jaffe, J.H., and Dax, E.M. Cognitive function and EEG testing 
in volunteer men inhaling volatile nitrites. Submitted Psvchopharmacologv . June. 1990. 

London, ED.. Cascella, N.G.. Wong, D.F., Phillips, R.L., Dannels, R.F., Links, J.M.. Herning, R., 
Grayson, R., Jaffe, J.H., and Wagner, H.N. Cocaine-indiced reduction of glucose utilization in human 
brain: A study using Positron Emission Tomography and FDG. Archives of General Psychiatry . 47, 567- 
576, 1990. 

Herning, R.I., Glover, B.J., Koeppl, B., Weddington, W., and Jaffe, J.H. Cognitive deficits in abstaining 
cocaine abusers. In: Residual Effects of Abused Drugs (J. Spenser and J. J. Boren, Eds.) National 
Institute on Drug Abuse Monograph Series, 101:167-178, 1990. 

London, E.D., Broussolle, E.P.M., Links, J.M., Wong, D.F., Cascella, N.G., Dannels, R.F., Sono, M., 
Heming, R., Snyder, F.R., Rippetoe, L.R., Toung, T.J.K., Jaffe, J.H., Wagner, H.N. Morphine-induced 
metabolic changes in the brain: Studies with Positron Emission Tomography and FDG. Archives of 
General Psychiatry . 47, 73-81, 1990. 

Herning, R.I., Glover, B.J. and Henningfield, J.E. Attention deficits during nicotine abstinence. 
Psvchopharmacologv . Submitted Jan., 1989. 

Pickworth, W.B., Herning, R.I., Koeppl, B. and Henningfield, J.E. Atropine-induced changes in 
spontaneous electroencephalogram in human volunteers. Military Medicine . 155: 166-170, 1990. 

Newlin. D.B., & Thomson, J.B. ( 1990). Alcohol challenge with sons of alcoholics: A critical review and 
analysis. Psychological Bulletin . 108, 383-402. 

Newlin, D.B., Byme, E.A., & Porges, S.W. (1990). Vagal mediation of the effect of alcohol on heart 
rate. Alcoholism: Clinical and Experimental Research . 14, 421-424. 

Newlin, D.B., Pretorius, MB. ( 1990). Sons of alcoholics report greater hangover symptoms than sons of 
nonalcoholics: A pilot study. Alcoholism: Clinical and Experimental Research, 14, 713-716 

Newlin, D.B., & Pretorius, M.B. (1991). Prior exposures to the laboratory enhance the effect of alcohol. 
Journal of Studies on Alcohol , in press. 

Newlin, D.B.. & Thomson. J.B. (1991). Chronic tolerance and sensitization to alcohol in sons of 
alcoholics. Alcoholism : Clinical a od Experimental Research , in press. 

Newlin. D.B., & Pretorius, M.B. (1991). Nonassociative mechanisms in the development of preferences 
for alcoholic flavors: Differences between sons of alcoholics and sons of nonalcoholics. Addictive 
Behaviors , in press. 



208 



O'Hara. B.J., Smith, S.S., Persico, A.. Wang, K., Cutting, G.R., Newlin, D.B., Gorelick, D.A., Uhl, G.R. 
(1991 ). Dopmaine D2 receptor alleles in substance abusers: Confounding effect of race, submitted. 



Published Abstracts 

Pretorius, M.B., Wong, C.J., & Newlin, D.B. (1990). Cardiovascular components of the response to 
morphine. Committee on Problems of Drug Dependence. 

Newlin, D.B.. Pretorius, MB., Wong, C.J., & Dax, E. (1990). Acute marijuana smoking reduces vagal 
tone. Committee on Problems of Drug Dependence. 

Uhl. G. Newlin, D.B., Pretorius. M.B., Park, J., & Cone, E. (1990). Antagonist-withdrawal up- 
regulalion of endogenous opiate antinociceptive systems. Committee on Problems of Drug Dependence. 

Hickey, J.E., Suess, P.E., Spurgeon, L., Newlin, D.B., & Porges, S.W. (1991). Vagal tone and attention 
in 8 to 12 year old males exposed to opiates in utero: A preliminary report. Committee on Problems of 
Drug Dependence. 

Newlin,, D.B., Wong, C.J., Pretorius. MB.. & Muntaner, C. (1991). Alcohol ingestion increases self- 
report of cocaine-craving: Individual differences in craving. Alcoholism: Clinical and Experimental 
Research . 12. 365. 

Pretorius, MB., Newlin, D.B., Wong, C.J., & Better, W.E. (1991). Individual differences in cocaine- 
craving: Physiological and affective correlates. Committee on Problems of Drug Dependence. 

Newlin. D.B., Pretorius, M.B., Wong, C.J., Stapleton, J.M., & London, E.D. (1991). Acute intravenous 
cocaine reduces cardiac vagal tone in cocaine abusers. Committee on Problems of Drug Dependence. 

Dax, E.M.. Newlin, D.B., Better, W.E., Wong, C.J., & Pretorius, M.B. (1991). Intravenous morphine 
produces initial heart rate increases and cardiac vagal tone decreases. Committee on Problems of Drug 
Dependence. 

Uhl, G.R., O'Hara, B.J., Smith. S.S., Persico, A., Wang, K., Cutting, G.R., Newlin, D.B., & Gorelick, 
D.A. (1991). Dopamine D2 receptor alleles in substance abusers: Confounding effect of race. 
Committee on Problems of Drug Dependence. 

Smith, S.S., Newlin, D.B., Uhl, G.R. (1991). Reliability and validity of the ARC Drug Use Survey. 
Committee on Problems of Drug Dependence. 

Heming, R.I.. Koeppl, B., Pickworth, W., Johnson, R.E., Fudala, J.H..Khazan, N. EEG Characteristics of 
burprenorphine maintenance and withdrawal in former heroin dependent subjects. Submitted to Society 
for Neuroscience 1991 

Heming, R.I., Glover, B.J., Koepple, B., Reddish, R., and London, E.Effects of Cocaine and Cocaine 
Withdrawal on the CNS: EEG, Evoked Potential and Performance. Fifth World Congress of Biological 
Psychiatry, Florence, Italy, June 9-14, 1991 

Heming, R.I., Brigham, J., Stitzer, M.L., Glover, B.J., Pickworth, W.B.and Henningfield, J.E. The 
effects of nicotine on information processing: Mediating a deficit. Society for Psychophysiological 
Research, Boston, MA., Oct., 1990 

Herning, R.I.. Glover, B.J., Koeppl, B., Reddish, R., Levin, R, Dax, E. Cognitive Deficits during cocaine 
abstinence persist. Committee on Problems of Drag Dependence, Richmond, June, 1990 



209 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00500-01 VL 

Period Covered: January 1. 1990 to December 31. 1990 

Title of Project: Individual Differences in Cocaine Craving: Physiological and Affective Correlates. 

Principal Investigators: Cooperating Units 

Newlin, D.B., Pretorius. M.B., Wong, C.J., & Better, W.E. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center. National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: 2 Professional: 1 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

We studied continuous self-report of cocaine-craving, autonomic measures (heart rate, cardiac vagal 
lone index, general motor activity, finger, and cheek temperature) and self-reported affect (POMS) in 
20 male research volunteers with heavy cocaine abuse histories. All subjects were exposed to a cocaine 
videotape and audiotape in counter-balanced order with a control audio and videotape. The autonomic 
correlates of craving induced by the cocaine-stimuli were small and could be attributed to nonspecific 
psychophysiological responding while attending to an external stimulus. However, autonomic and 
affective correlates emerged when we correlated overall levels of cocaine- craving with pre-stimulus 
heart rate and self-reported affect. Replicating our preliminary study with a smaller number of 
residential volunteers, cocaine-craving was negatively correlated (r = -.45, p<05) with resting (i.e., pre- 
stimulus) heart rates; craving was also negatively correlated (r = -.44, p<.06) with baseline forehead 
temperature. Cocaine-craving was positively correlated with both positive and negative affect on the 
POMS. These results indicated stable trait characteristics rather than reactions to the tapes. There were 
interesting parallels between the autonomic patterns of the higher cocaine-craving subjects and "fussy," 
irritable, dysregulated infants. Cocaine abusers who do not crave cocaine may have blunted affect. 
Cocaine- craving was much greater in this outpatient study than our otherwise similar residential study, 
suggesting that drug availability or anticipation may determine the intensity of cocaine-craving. Unlike 
the preliminary study, frequency (but not quantity) of cocaine use was positively correlated (r = .70, p 
<001) with cocaine-craving. These results indicate that stable individual physiological correlates of 
cocaine-craving may be more robust than evoked responses to cocaine-stimuli in this paradigm. 



210 



Publications 

Pretorius, M.B., Newlin, D.B.. Wong, C.J., & Better, W.E. (1991). Individual differences in cocaine- 
craving: Physiological and affective correlates. Committee on Problems of Drug Dependence. 



211 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 10701-02 VL 

Period Covered: January 1. 1990 to December 31, 1990 

Title of Project: Alcohol Ingestion Increases Self-Report of Cocaine-Craving: Individual Differences in 
Craving 

Principal Investigators: Cooperating Units 

Newlin, D.B.. Wong, C.J., Pretorius, M.B., & Muntaner, C. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 3 Professional: 1 Other: 2 

Check Appropriate Boxes: 

_X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Cocaine abusers often report difficulty abstaining from cocaine while intoxicated with alcohol, and 
alcoholism and cocaine dependence have strong comorbidity. Therefore, we sought to determine 
whether alcohol intoxication would increase cocaine-craving induced by exposure to videotapes of an 
individual self-administering cocaine compared to a control tape of an individual painting. Subjects were 
12 male residential volunteers with histories of both heavy alcohol and cocaine use. They drank water 
the first day, and a high (1.1 g/kg), moderate (0.64 g/kg), placebo (0.0 g/kg) dose of alcohol, or water on 
4 separate (approx. alternate) days in pseudo- randomized order in double-blind fashion. Continuous 
self-report of desire for cocaine and autonomic measures were recorded before drinking, and during and 
after watching the stimulus tapes in the rising and falling BAC curves. 

Although mean levels of cocaine-craving were not intense, subjects reported significantly more cocaine- 
craving after ingestion of alcohol; this effect was nonsignificantly greater in the rising BAC curve when 
BAC was highest. We then divided subjects into 7 who craved cocaine and 5 with minimal or no 
craving during the entire experiment; these two groups had equivalent cocaine and alcohol histories. 
High cocaine-cravers had trait characteristics of significandy lower resting heart rate, higher vagal tone 
index, and greater general motor activity on a stabdometer under their chairs both before and after 
drinking. Lower resting heart rates among high cocaine-cravers was replicated in a second 
(nonresidential) study with 20 cocaine abusers. The autonomic pattern for high cocaine-cravers parallels 
that of "fussy," irritable, dysregulated infants. The results suggest a role for alcohol intoxication in 
craving for cocaine. They also underscore the potential importance of individual differences in drug- 
craving among users with equivalent abuse histories. 



212 



Publications 

Newlin, D.B., Wong, C.J., Pretorius, M.B., & Muntaner, C. (1991). Alcohol ingestion increases self- 
report of cocaine-craving: Individual differences in craving. Alcoholism: Clinical and Experimental 
Research . 12, 365. 



213 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00501-01 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Intravenous Morphine Produces Initial Heart Rate Increases and Cardiac Vagal Tone 
Decreases. 

Principal Investigators: Cooperating Units 

Newlin. D.B., Better, W.E., Wong, C.J., Pretorius, M.B., & Dax, E.M. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 1 Professional: 0.25 Other: 0.75 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

We have found that many abused drugs increase heart rate (HR) and decrease cardiac vagal tone index 
(V). Traditional views of the limited effect of the opiates on the cardiovascular system would suggest 
that the opiates represent an exception to this observation. However, this may be due to recordings 
taken after compensatory mechanisms have stabilized cardiovascular parameters. Therefore, we 
recorded HR and V continuously before, and for 30 min after i.v. morphine (15 mg) injection to 
measure initial cardiovascular responses. V is a well-validated measure of vagal inhibition of the heart 
based on time series analysis of HR variability entrained with respiration (i.e., respiratory sinus 
arhythmia). The subjects were 9 opiate abusers who were research volunteers living on a residential 
ward. Intravenous morphine produced immediate HR increases (p<.01) of +9 bpm, and V decreases 
(p<.05) of -1.4 log units. These responses gradually returned toward baseline during the 30 min after 
injection. These effects were in the same direction, but of larger magnitude than our previous results 
with i.m. morphine (20 mg). Therefore, morphine may not be an exception to the pattern of 
cardiovascular responding to various abused drugs when considering initial responses before 
compensatory mechanisms develop. Moreover, these results indicate that the HR response to morphine 
has a substantial parasympathetic component. 



214 



Publications 

Dax. E.M., Newlin, D.B., Better, W.E.. Wong, C.J., & Pretorius, M.B. Intravenous morphine produces 
initial heart rate increases and cardiac vagal tone decreases. Presented at Committee on Problems of 
Drug Dependence, Palm Beach, Florida, 1991. 



215 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00502-01 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Cardiovascular Responses to Naloxone-Precipitated Withdrawal: A Test of a 
Hypothesis Concerning Drugs of Abuse. 

Principal Investigators: Cooperating Units 

Newlin, D.B., Wong, C.J., Better, W.E., and Cheskin, L. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 125 Professional: 0.25 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

We have examined cardiovascular responses in humans to a wide range of abused drugs, including 
cocaine, marijuana, alcohol, nicotine, methylphenidate, morphine, dilaudid, and pentobarbitol. All these 
drugs increase heart rate and decrease vagal tone, particularly cocaine and marijuana. Vagal tone 
measures parasympathetic influences on the heart noninvasively by quantifying respiratory sinus 
arrhythmia, or heart rate variability entrained with respiration. Withdrawal of vagal inhibition produces 
tachycardia (i.e., increased heart rate). We have proposed that stimulation of the mesolimbic dopamine 
reward system by these abused drugs produces parallel activation of locomotor activity (Wise & Bozarth, 
1987) and vagally-mediated tachycardia. This raises the possibility that measuring parasympathetic 
withdrawal and its associated tachycardia may prove useful as a simple index of a drug's abuse liability. 
This would require that psychoactive drugs that are not abused do not also show the same pattern (i.e., 
decreased vagal tone and increased heart rate). 

The purpose for this study was to determine whether the tachycardia sometimes produced by naloxone- 
precipitated withdrawal from opiates is mediated by decreased vagal tone. We hypothesized that this 
tachycardia is sympathetically rather than parasympathetically mediated because naloxone is not an 
abused drug. 

We administered 0.4 mg intramuscular naloxone to 12 opiate users who were classified as opiate- 
dependent on the basis of clinical history and toxicology. All 12 subjects exhibited signs of naloxone- 
precipitated withdrawal, as indicated by elevated scores on the Opiate Withdrawal Scale. We recorded 
heart rate and vagal tone continuously before and for 30 min after the intramuscular injection of 
naloxone. The cardiovascular response peaked from 11 to 16 min after the injection. We preselected the 
8 subjects with the greatest heart rate increases in order to provide the strongest possible test of the 
hypothesis concerning vagal tone (i.e., vagal tone would not be expected to decrease if heart rate did not 
increase). For these 8 subjects, heart rate increased significantly (F(1.7)=20.3, p< 



216 



005) approx. 7 to 8 beats/min, and vagal tone decreased slightly but nonsignificantly F(l,7)=4.4, n.s.). A 
lower frequency (approx. 0.10 Hz) rhythm in the heart rate variability spectrum increased 
nonsignificantly (F(l,7)=3.0, n.s.). 

This pattern of cardiovascular results was very different from that of any abused drug that we have 
assessed (listed above). We interpreted this pattern to indicate that, as predicted, the tachycardia from 
naloxone-precipitated withdrawal was sympathetically as opposed to parasympathetically mediated. 



Publications 

Newlin, D.B., Wong, C.J., Better, W.E., & Cheskin, L.J. Cardiovascular Responses to Naloxone- 
Precipitated Withdrawal: An Exclusionary Test of a Hypothesis Concerning Drugs of Abuse. Paper 
presented at the Association for the Advancement of Behavior Therapy, 1991. 



217 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00504-01 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Acute Intravenous Cocaine Reduces Cardiac Vagal Tone in Cocaine Abusers. 

Principal Investigators: Cooperating Units 

Newlin, D.B., Pretorius, MB., Wong, C.J., Stapleton, J.M., & London, E.D. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.5 Professional: 0.5 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Researchers have assumed that the pronounced tachycardia produced by cocaine is due to sympathetic 
autonomic mechanisms because cocaine blocks reuptake of catecholamines. We studied the acute 
effects of cocaine on parasympathetic mechanisms to determine if there was a significant vagal 
component to this tachycardia. We used a noninvasive measure of cardiac vagal tone. This measure, 
which has been well-validated in animals and human subjects, is based on time series analysis of 
successive R - R intervals. Vagal tone index quantifies heart rate variability in the same frequency 
band as respiration (i.e., respiratory sinus arrhythmia). We considered vagal blockade impractical to 
resolve this issue because it produces very large baseline shifts in heart rate. We administered cocaine 
(20 mg and 40 mg) and placebo (saline) intravenously on separate days in pseudo-randomized order in 
double-blind fashion to 10 male residential volunteers with histories of cocaine abuse. Cocaine 
produced dose-dependent increases in heart rate. The effect was precisely mirrored by robust decreases 
in vagal tone index, as well as decreases in a lower frequency heart rate rhythm associated with blood 
pressure homeostasis. Injection of saline (i.e., cocaine cues) produced an initial 14 bpm increase in 
heart rate that had no significant vagal component. Vagal tone index and the lower frequency rhythm 
decreased approximately 2 to 2.5 log units in response to 40 mg cocaine, with a trough 7 to 14 min after 
intravenous administration. Therefore, cocaine produced a pronounced decrease in heart rate 
variability. The results indicate that cocaine-induced tachycardia has a strong parasympathetic 
component. Further research is needed to determine whether the vagal and sympathetic effects of 
cocaine on the autonomic nervous system are additive or synergistic. The findings have implications 
for a better understanding of the cardiotoxicity of cocaine. 



218 



Publications 

Newlin, D.B.. Pretorius, MB., Wong, C.J., Stapleton, J.M., & London, ED. (1991). Acute intravenous 
cocaine reduces cardiac vagal tone in cocaine abusers. Paper presented at Committee on Problems of 
Drug Dependence, Palm Beach, Florida, 1991. 



219 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00505-01 VL 

Period Covered: January 1. 1990 to December 31, 1990 

Title of Project: 

The ARC Drug Expectancy Questionnaire: An Instrument for Assessing Expectancies Concerning Use 
of Cocaine, Heroin. Marijuana, Alcohol, and Nicotine. 

Principal Investigators: Cooperating Units 

Wong, C.J., Newlin, D.B.. Better, W.E., & Pretorius, M.B. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.10 Professional: 0.10 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

In the alcohol literature, researchers have found that cognitive expectations about how alcohol will 
affect people are potent predictors of individual alcohol use. These expectations tend to be stronger 
predictors of drinking than other variables, such as demographics or familial history of alcoholism. 
People who expect, for example, that alcohol will make them happy, friendly, attractive, etc., drink 
more alcohol than people who expect that alcohol will make them clumsy, unattractive, and sleepy. 
These cognitive expectations can be measured in primary school children, and appear to be a product of 
both societal attitudes toward alcohol intoxication and individual experiences with the drug. 

We developed the Addiction Research Center Drug Expectancy Questionnaire (ARCDEQ) to extend 
this research to drugs other than alcohol. We included tobacco as a comparison or control drug because 
we thought it unlikely to show the magnitude of subjective expectations associated with the other drugs. 
The ARCDEQ has 46 items for each of five drugs (i.e., alcohol, cocaine, heroin, marijuana, and 
tobacco). Thirty six of the items referred to expectations about acute effects of the drug, such as feeling 
"anxious" or feeling "bold." These items were balanced to include those on each end of continuums 
represented by "good" vs. 'had" effects, stimulant vs. depressant effects, enhancing vs. impairing 
effects, and prosocial vs. hostile effects. Ten items were also included for each drug to assess longterm 
expectations about how chronic use will affect them, such as "enjoy life more" or "get sick." Each item 
used a Likert scale with descriptive anchors of "unlikely" to "likely", with "neither" in the center of the 
scale. 

We have administered this questionnaire in computerized form to 86 abstinent, light, and heavy drug 
users. To date, 75 men and 1 1 women have answered the ARCDEQ. Twenty subjects answered the 
questionnaire twice to assess test-retest reliability. For each drug, we divided subjects based on 



220 



whether (hey were naive, light, or heavy users of that particular drug, based on quantity-frequency 
items from the ARCDEQ itself, and the ARC Drug Use Survey, administered separately. 

We will report on preliminary analyses of this data in which we eliminated some items and developed 
empirically-derived scales for data reduction and reporting. We consider expectations about abused 
drugs to be important for two reasons. 

First, they suggest motives for excessive drug use and may predict who escalates from experimental to 
heavy use. Second, they have direct relevance for prevention and psychological treatment of drug 
abuse, namely, to develop interventions to change these expectancies. 



Publications: 

Wong, C.J.. Newlin. D.B., Better, W.E., & Pretorius, M.B. The ARC Drug Expectancy Questionnaire: 
An Instrument for Assessing Expectancies Concerning Use of Cocaine, Heroin, Marijuana, Alcohol, 
and Nicotine. 



221 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00506-01 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Acute Effects of Various Abused Drugs on Heart Rate and Cardiac Vagal Tone: A 
Common Factors Approach. 

Principal Investigators: Cooperating Units Newlin, D.B., Pretorius, M.B., Wong, C.J., Better, W.E., 
& Pickworth, W.B. 

Lab/Branch: Etiology Branch 

Section: Vulnerability Laboratory 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.5 Professional: 0.5 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Drug-induced cardiovascular responding appears to be a common factor among abused drugs that may be 
linked to psychomotor stimulant activation and increased mesolimbic dopamine levels. Therefore, the 
pattern of cardiovascular responses to abused drugs may reflect their reward value. We measured heart 
rate (HR) and vagal tone index (V) before, 60 and 120 min after double-blind administration of low and 
high dosages of smoked marijuana, or oral alcohol, hydromorphone, pentobarbital, and d- amphetamine. 
There were two triple-dummy (i.e., smoke, drink, and capsule) placebo sessions. The 12 sessions were in 
randomized order on approx. alternate days. The subjects were 9 male poly-drug abusers who were 
residential volunteers. V is a well-validated measure of parasympathetic inhibition of the heart that 
quantifies respiratory sinus arrhythmia using time series analysis, i.e., beat-to-beat variability in HR that 
is entrained with respiration. Marijuana, alcohol, and pentobarbital increased HR and decreased V 
relative to placebo in a dose-dependent manner, these results replicate our previous findings with 
marijuana and alcohol. D-amphetamine increased HR relative to placebo only at the low dose; it 
appeared to fail to increase HR at the high dose because of baroreceptor mechanisms associated with 
blood pressure increases. Hydromorphone had no effect on HR or V at 60 or 120 min after drug 
ingestion. We have shown that HR increases and V decreases occur immediately after opiate 
administration, and are well compensated by 60 or 120 min. These results suggest some commonality in 
the cardiovascular responses to various abused drugs, and may reflect the common excitatory aspects of 
these drugs. However, cardiovascular responses during the rising blood drug curves may better illustrate 
this pattern because they occur before compensatory mechanisms develop. 



222 



Publications 

Newlin. D.B., Pretorius. MB., Wong, C.J.. Better, W.E., & Pickworth, W.B. Paper presented at the 
Committee on Problems of Drug Dependence, Palm Beach, Florida, June, 1991. 



223 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 09601-03 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Antagonist- Withdrawal Up-Regulation of Endogenous Opiate 

Principal Investigators: Cooperating Units Uhl, G., Newlin, D.B., & Pretorius, M.B. 

Lab/Branch: Etiology Branch 

Section: Vulnerability Laboratory 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.5 Professional: 0.5 Other: 1 

Check Appropriate Boxes: 

_X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Changes in expression of the principle opioid peptide gene, preproenkephalin, follow stimulation or 
inhibition of inputs to antagonist drugs. The duration of proenkephalin gene up- or down-regulation can 
outlast the duration of the stimulus, suggesting that regulated gene expression could provide one substrate 
for storing information about previous stimulus or drug exposure. This would have direct implications 
for possible mechanisms of opiate tolerance and withdrawal. We have thus sought evidence for 
functional up-regulation of enkephalinergic gene regulation systems in humans. The subjects were 19 
males with no reported histories of opiate abuse. Pain self-report to a 5 min ice-water immersion and 
cold-stress-induced analgesia were tested before 8 days of antagonist treatment, 2 days after the last dose 
of antagonist (when excretion of antagonist metabolites was finished) or placebo, and finally 30 min after 
an acute oral dose of naloxone. Self-report of pain to the initial stages of the ice-water immersion were 
significantly reduced in subjects 2 days after opiate antagonist treatment. Initial results of the study 
provide evidence for increased function in endogenous opioid systems after antagonist washout. Current 
studies aim to separate pre- and post-synaptic components to this effect. 

We studied 30 normal males in a second study that was a replication and extension of the first. The 
procedure was similar, with the addition of an oral hydromorphone challenge at the end to: 1) quantify 
the degree of analgesia to a mu-type opiate and 2) rule out post-synaptic changes as an explanation of the 
earlier results. 



224 



Publications 

Uhl, G.R., Newlin, D.B., Pretorius, M.B., Park, J., & Cone, E. (1990). Antagonist-withdrawal up- 
regulation of endogenous opiate antinociceptive systems. Paper presented at Committee on Problems of 
Drug Dependence, Palm Beach, Florida, 1991. 



225 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00507-01 VL 

Period Covered: January 1. 1990 to December 31, 1990 

Title of Project: Vagal Tone and Attention in 8 to 12-year-old Males Exposed to Opiates in Utero: A 
Preliminary Report. 

Principal Investigators: Cooperating Units Hickey, J.E., Suess, P.E., Spurgeon, L., Newlin, D.B., & 
Porges, S.W. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore. MD 21224 

Total Man Years: 1.5 Professional: 0.5 Other: 1 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

We assessed vagal tone changes during an attention demanding CPT task in 12 boys (mean age 9 yrs, 5 
mos) exposed prenatally to opiates and 12 (mean age 9 yrs, 10 mos) male controls. Mothers of opiate- 
exposed and control boys were primarily single and of lower income. Groups did not differ significantly 
on mother's education, income, or marital status. Racial composition of the groups also did not differ 
(opiate-exposed: 8 black, 4 white, controls 7 black, 5 white). Vagal tone was measured pre- and post- 
baseline and during the 3 tasks of the Gordon Diagnostic System. Vagal tone is a heart rate variability 
measure that quantifies parasympathetic inhibition of the heart. Results indicated that opiate-exposed 
boys failed to suppress vagal tone compared to control boys when distractors were added to a vigilance 
task (p .05). In normal children and adults, vagal tone is suppressed during tasks requiring sustained 
attention. These preliminary results indicate that normal physiological responses to increased attentional 
demand may be impaired in opiate-exposed boys in this age range. These physiological response patterns 
were not associated with prenatal alcohol, nicotine, or marijuana exposure in these samples. The design 
of this study does not distinguish among genetic, teratogenic, or child-rearing practice effects. Further 
research is needed to replicate and extend these findings as a possible risk factor for subsequent drug 
abuse in children exposed prenatally to opiates. 



226 



Publications 

Hickey, J.E., Suess, P.E., Spurgeon, L., Newlin, D.B., & Porges, S.W. Vagal tone and attention in 8 to 
12 year old males exposed to opiates in utero: A preliminary report. Paper presented at Committee on 
Problems of Drug Dependence, Palm Beach, Florida, 1991. 



227 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 06801-04 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Cognitive Neurophysiologic Signs of Cocaine Abstinence 

Principal Investigators: Cooperating Units Herning, R.I., 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 1.60 Professional: .40 Other: 1.20 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Cognitive impairments and sleep disruption have been reported in patients withdrawing from cocaine. 
The nature of these disorders have yet to be documented in clinical laboratory studies. The present study 
evaluates cognitive information processing in subjects on a clinical ward withdrawing from cocaine with 
a battery of tasks (auditory rare event monitoring, two continuous performance tasks, four Stemburg 
memory tests ). Sleep quality and duration is monitored by a subjective questionnaire. Fourteen subjects 
including controls have been tested in this study over a 6 to 8 week withdrawal period. Stimulus 
evaluation and memory deficits were observed in the cocaine addicts. 

Clarification of the nature of the cognitive deficits and of sleep loss will lead to more effective treatment 
strategies for cocaine withdrawal. 



228 



Publications 

Heming, RE., Glover. B.J., Weddington, WW., Koeppl, B.S.. and Jaffe, J.H. Cognitive decrements 
during cocaine abstinence were not related to depression. Biological Psychiatry . Submitted January, 
1990, in revision. 

Weddington, W.W., Brown, B.S., Haertzen, C.H., Cone, E.J., Dax, E.M., Heming, R.I., and Michaelson, 
B.S. Changes in mood, craving and sleep during acute abstinence reported by male cocaine addicts: A 
controlled residential study. Archives of General Psychiatry . 47: 861-868, 1990. 

Heming, R.I., Glover. B.J., Koeppl, B., Reddish, R., Levin, F., Dax, E. Cognitive deficits during cocaine 
abstinence persist. Paper presented at Committee on Problems of Drug Dependence, Palm Beach, 
Florida, June. 1991. 

Heming, R.I., Glover, B.J.. Koeppl, B., Weddington, W., and Jaffe, J.H. Cognitive deficits in abstaining 
cocaine abusers. In: Residual Effects of Abused Drugs (J. Spenser and J.J. Boren, Eds.) National 
Institute on Drug Abuse Monograph Series, 101: 167-178, 1990. 

Herning, R.I.. Glover, B.J., Koeppl, B., Reddish, R., and London, E. Effects of Cocaine and Cocaine 
Withdrawal on the CNS: EEG, Evoked Potential and Performance. Fifth World Congress of Biological 
Psychiatry, Florence, Italy, June 9-14, 1991. 



229 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 05801-04 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Mapping the Effects of Cocaine by EEG 

Principal Investigators: Cooperating Units Heming, R.I.. London, E., & Stapleton, J. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .40 Professional: .20 Other: .20 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

The effects of cocaine on scalp EEG and FDG PET scans are being compared to determine the brain 
areas involved in the cocaine-induced euphoria. In previous studies, cocaine increased EEG beta power. 
The distribution of cortical areas responsible for the EEG beta increase and the time course of the beta 
increase have not as yet been determined. The present study was designed to answer these two questions. 

The complimentary nature of the EEG and PET data will delineate the anatomical and electrophysiologic 
mechanisms involved in cocaine induced euphoria. 

Twenty subjects were tested using EEG measures with placebo, 20m g and 40m g of cocaine in double 
blind order in previous years and seven additional subjects were tested during the current year. EEG beta 
increased in dose dependent manner starting immediately after the injection and continuing for twenty 
minutes. The increase in EEG beta was maximal in frontal cortical areas. The relationship between the 
increase in beta and subjective state is currently being investigated. 



230 



Publications 

London, ED., Cascella, N.G., Wong, D.F., Phillips, R.L., Dannels, R.F., Links, J.M., Heming, R.I., 
Grayson, R., Jaffe, J.H.. and Wagner, H.N. Cocaine-induced reduction of glucose utilization in human 
brain: A study using Positron Emission Tomography and FDG. Archives of General Psychiatry . 47, 567- 
576, 1990. 



231 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 02101-05 VL 

Period Covered: January 1, 1990 to December 31. 1990 

Title of Project: Acute Abstinence from Tobacco: Electrophysiological and Cognitive Signs 

Principal Investigators: Cooperating Units Heming, R.I., Henningfield, J., & Pickworth, W.B. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 21224 

Total Man Years: .025 Professional: .025 Other: 0.0 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

_ Minors 

Interviews 

Summary of Work 

The laboratory's efforts were directed toward the quantification of the cognitive and performance deficits 
during nicotine withdrawal and the treatment of these deficits with nicotine chewing gum. The EEG, 
cognitive, and cognitive process was monitored during a ten-day period of tobacco withdrawal in heavy 
smokers. Some of the changes persisted over the entire ten-day deprivation period. These measures 
included EEG alpha frequency, theta power, performance on selected cognitive tasks (especially a rapid 
arithmetic task) and a cognitive event related potential measure (N100 amplitude). Stimulus evaluation 
time, as measured by P300 latency, and the depth of stimulus evaluation battery were affected early 
during the tobacco deprivation period, but returned to smoking levels later during the deprivation period. 
Thus, the cognitive deficits are clearly apparent during abstinence from tobacco and contribute to relapse 
during treatment. The deficits during withdrawal have at least two different components - one affecting 
stimulus evaluation which dissipates after 5 to 7 days of abstinence and one affecting attention 
accompanied by lower arousal which persists ten days or longer. During the year this data was analyzed 
and two papes were submitted for publication. 



232 



Publications 

Heming, R.I., Glover. B.J., and Henningfield, J.E. Attention deficits during nicotine abstinence. 
Psvchopharmacologv . Submitted January. 1989. 

Heming, R.I., Brigham, J., Stitzer, M.L., Glover, B.J., Pickworth, W.B., and Henningfield, J.E. The 
effects of nicotine on information processing: Mediating a deficit. Society for Psychophysiological 
Research. Boston, Massachusetts, October, 1990. 



233 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 02001-05 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Mapping the Effects of Opioid Agonists by EEG 

Principal Investigators: Cooperating Units Heming, R.L, London, E.D., Stapleton, J., & Philips, R. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .025 Professional: .025 Other: 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

_ Minors 
Interviews 

Summary of Work 

Effects of morphine on the scalp EEG and FDG PET scans are being compared to determine the brain 
areas invoked in euphoria. Etiology collected in past years and is now analyzing the EEG data from 20 
scalp locations from post addicts receiving placebo, 15 and 30 mg injections of morphine. These subjects 
subsequently received FDG PET scans while receiving placebo and 30 mg of morphine. The PET scans 
are performed by our collaborators. The EEG data by itself provides insight into time course of 
electrophysiologic effects of a mu agonist in humans and the cortical distribution of mu effects. PET 
techniques do not by themselves provide information about the time course of the mu effects. In the 
preliminary analysis, twelve subjects had increased EEG delta and the theta power beginning 15 minutes 
and persisting until 45 minutes after the intramuscular injection. Changes in artifact detection were being 
investigated so that the relationship between these EEG changes and subjective effects of morphine can 
be investigated 



234 



Publications 

London, E.D.. Broussolle, E.P.M., Links, J.M., Wong, D.F., Cascella, N.G., Dannels, R.F., Sono. M., 
Heming, R., Snyder, F.R., Rippetoe. L.R., Toung, T.J.K., Jaffe, J.H., Wagner, H.N. Morphine-induced 
metabolic changes in the brain: Studies with Positron Emission Tomography and FDG. Archives of 
General Psychiatry . 47, 73-81, 1990. 



235 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 03101-05 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Effects of Atropine on Cognitive Information Processing 

Principal Investigators: Cooperating Units Heraing, R.I. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: .025 Professional: 025 Other: 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

_ Minors 
Interviews 

Summary of Work 

An extensive battery of sensory and cognitive electrophysiological tasks is used to assess sensory, 
cognitive and performance deficits produced by atropine. The tasks include eyes open and eyes closed 
EEG, brainstem auditory evoked response, pattern reversal visual evoked response, the auditory rare 
event monitoring task, auditory continuous performance task and Sternberg auditory memory task (both 
immediate and delayed). Each of four doses of atropine (0, 2, 4 and 6 mg/70 kg) is investigated on two 
occasions. Eight subjects have been tested on these procedures. 

The purpose of the study is to better understand the effects of cholinergic agents on cognition and 
performance; in particular, where in the information processing sequence atropine exerts its effects. The 
EEG and evoked response data have been reported in military and scientific journals. Atrophine at doses 
4 mg or greater increase EEG slowing and reduces cognitive evoked potentials and performance. The 
EEG results were published and evoked potential analysis began over the last year. 



236 



Publications 

Pickworth, W.B., Herning. R.I., Koeppl, B. and Henningfield, J.E. Atropine-induced changes in 
spontaneous electroencephalogram in human volunteers. Military Medicine . 155: 166-170, 1990. 



237 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00508-01 VL 

Period Covered: January 1, 1990 to December 31, 1990 

Title of Project: Neurocognilive Status of Young Boys Exposed to Opiates in Utero 

Principal Investigators: Cooperating Units Herning, R.I., Spencer, J., and Guo, X. 

Lab/Branch: Etiology Branch 

Section: 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 70 Professional: .70 Other 0- 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

X Minors 
Interviews 

The study seeks to detennine whether boys 8 to 12 who were exposed to opiates in utero, have 
any neurocognitive deficits as compared to similar aged control boys. Two control groups were 
employed in this study. BoUi groups were not exposed to opiates in utero. The boys in one 
group lived with an opiate using mother and the boys in the other group did not. The boys 
exposed in utero to opiates have increased intentional tremor and appear to have alterations in 
event related potentials. The study is ongoing and about 30 boys have been tested. 



238 



Treatment Branch 

David Gorelick, M.D., Ph.D., Chief 

Overview 

The Treatment Branch conducts research on the efficacy and safety of existing and developing 
pharmacologic and psychosocial treatments for drug abuse, including the influence of treatment on the 
biomedical consequences of drug abuse. Current research focuses on cocaine and opiate abuse. Research 
is carried out on both the ARC residential unit and the outpatient clinic. The Branch also collaborates, as 
appropriate, with other clinical facilities in the Baltimore area. The Treatment Branch is organized into 
the Pharmacotherapy Laboratory (Daviu A. Gorelick, M.D., Ph.D., Chief). Clinical Trials Laboratory 
(Paul Fudala, Acting Chief), and Drug Abuse Treatment Evaluation Unit (John Ball, Ph.D., Chief). 

The Branch studies new treatments with a view toward developing effective strategies for use by the 
clinical treatment community. Potential therapeutic medications are chosen for study on several 
grounds: 1) their existing clinical use, even in the absence of controlled scientific data, 2) theoretical 
deduction from the known neuropharmacology of drugs of abuse, or 3) leads from other ARC or NIDA 
researchers. Detailed diagnostic and biopsychosocial characterization of subjects is done to identify 
predictors of treatment compliance and response, develop optimum matching of patients to treatment, 
and determine the extent to which biomedical and psychosocial consequences of drug abuse are affected 
by treatment. These objectives are pursued using a variety of research techniques, including ( 1 ) single- 
blind and double-blind, placebo-controlled designs with pharmacologic interventions; (2) experimental 
designs employing random assignment to control or comparison groups; (3) obtaining and quantifying 
observational data to clarify behaviors significant to the conduct of drug abuse treatment. Whenever 
possible, long-term follow-up is obtained on subjects after active treatment has ended, in order to assess 
the persistence of treatment effects. 

Multidisciplinary studies are frequently conducted in collaboration with other laboratories of the ARC, 
including the Chemistry and Drug Metabolism and Neuroendocrinology Laboratories (Clinical 
Pharmacology Branch) , Vulnerability Laboratory (Etiology Branch), and Molecular Neurobiology 
Laboratory (Neuroscience Branch). In addition, several clinical drug abuse programs of the Maryland 
Substance Abuse Administration, University of Maryland School of Medicine, and Department of 
Veterans Affairs have indicated a willingness to collaborate with the Branch. 

Long-term goals of the Branch will continue to be the exploration of scientific issues significant to the 
treatment process and the examination of interventions that have promise for improved treatment. 
Specific areas of interest include the role of neurotransmitter receptor ligands and anti-convulsants as 
potential therapeutic medications for drug abuse, the impact of HIV infection on drug abuse treatment, 
and concurrent psychiatric diagnoses in drug abusers. Many of these efforts are in cooperation with 
NIDA's Medication Development Division and AIDS program. 



1. Pharmacotherapy Laboratory - David Gorelick, M.D., Ph.D., Chief 
Overview 

The goal of the Pharmacotherapy Laboratory is the development of new pharmacologic treatments for 
drug abuse. Potential treatment medications are evaluated in a controlled residential environment using a 
variety of interdisciplinary experimental paradigms (e.g., drug self-administration) that allow study of 
clinically relevant mechanisms of action (both pharmacological and psychological), potential to produce 



239 



toxicity and/or adverse interactions with drugs of abuse, and the pharmacokinetics of the medication. 
Various behavioral, psychological, physiologic, and pharmacokinetic parameter are measured, including 
those performed in collaboration with other ARC laboratories, such as electrophysiological assessments 
and PET scanning. 

Summary of Current Research 

A. Effects of carbamazepine on cocaine self-administration 

The anti-convulsant medication carbamazepine has attracted much attention as a potential treatment for 
cocaine addiction, but there is little systematic data from controlled human studies on either its efficacy 
or the safety of its interactions with cocaine. This double-blind , double-dummy, placebo-controlled 
residential study evaluates the effect of targeted carbamazepine plasma levels on cocaine self- 
administration, self-reported cocaine craving, and conditioned responses to cocaine-associated stimuli in 
18 cocaine addicts. This study also evaluates the influence of carbamazepine on the subjective and 
cardiovascular effects of cocaine, and the pharmacokinetics of carbamazepine in cocaine addicts. 

B. Comparison of detoxification treatments for opiate addiction 

The purpose of this ongoing inpatient study is to compare the efficacy of buprenorphine to the alpha- 2- 
adrenergic agonist clonidine for the rapid detoxification of heroin addicts. A major focus of this study 
will be to determine whether a successful opiate detoxification can be accomplished by administering 
buprenorphine for no longer than 72 hours, since federal regulations allow for the use of an opiate for up 
to 72 hours for opiate-dependence treatment without special licensing requirements of the practitioner. 

C. HIV infection, high-risk behaviors, and drug abuse 

HIV infection is associated with intravenous drug use and high-risk behavior such as needle sharing, but 
relatively little is known about these variables in non-intravenous drug abusers or how these variables are 
influenced by drug abuse treatment. This study will evaluate the influence of drug of abuse and route of 
administration on HIV antibody status and high-risk behaviors, as well as the latter's influence on 
treatment compliance and outcome. These issues will be addressed both retrospectively, by analyzing 
data already collected on ARC subjects, and prospectively, by administering questionnaires or structured 
interviews to patients entering drug abuse treatment research protocols. 

D. Esterase activity in human cocaine abusers 

Plasma and RBC esterases are the chief metabolizing enzymes for cocaine in humans. Studies with other 
substrates indicate that these enzymes are under genetic control and show population variability, but there 
are no human studies of their effects on cocaine metabolism or cocaine abuse. This study, in 
collaboration with Dr. Raymond Woosley, Georgetown University, will measure plasma and RBC 
esterase activity in cocaine abusers and determine the influence of this metabolic parameter on response 
to cocaine and treatment outcome. 

E. Dopamine D2 receptor allelic linkage in substance abuse 

Recent research reports have suggested a possible association between a particular dopamine D2 receptor 
allele and alcoholism. The Treatment Branch is collaborating with the Molecular Neurobiology 
Laboratory in recruiting and diagnosing subjects for a study of the association between dopamine D2 
receptor allele and heavy peak lifetime use of a variety of drugs, including alcohol, cocaine, opiates, and 
nicotine. Possible confounding factors, such as racial background of subjects and possible drug use in 
control subjects, are also being addressed 



240 



PUBLICATIONS 

Ball, John C. "Opening the "Black Box" of Drug Abuse Treatment - Measurement and Evaluation of the 
Treatment Domain," Committee on Problems of Drug Dependence, Inc. (Abstracts), Fifty-Second Annual 
Scientific Meeting, June 10- 14, 1990. 

Ball. John C. "The Status of Methadone Maintenance Treatment in the Uiuted States," Australian 
Methadone Conference. Keynote Speaker (Sydney, Australia). July 9-10, 1990. 

Ball, John C. "Evaluating Methadone Maintenance Programs," Keynote Speaker Midwest Regional 
Methadone Conference, Milwaukee, Wisconsin, September 17-18, 1990. 

Ball, J.C. "A Comprehensive Evaluation of Methadone Maintenance Programs in New York City, 
Philadelphia and Baltimore," Advances in Alcohol & Substance Abuse (Guest Issue), (In press). 

Ball, J.C. "A Schema for Evaluating Methadone Maintenance Programs," In: L.S. Harris (ed.), NIDA 
Research Monograph: Proceedings of the 51st Annual Scientific Meeting of the Committee on Problems 
of Drug Dependence. 1989 NIDA Res. Monogr. 95:74-77, 1990. 

Ball. J.C. "The Effectiveness of Methadone Maintenance Treatment in the United States An Overview." 
Presented at the "What Works Conference" in New York, October 22-24, 1989. (In press). 

Ball, J.C. "The Similarity of Crime Rates Among Heroin Addicts in New York City, Philadelphia and 
Baltimore," In: R. Rachin (ed.), Journal of Drug Issues (Guest Issue), Fall 1990 (In press). 

Ball. J.C, Ross A., and Jaffe J.H., "Cocaine and Heroine Use by Methadone Maintenance Patients." 
NIDA Res. Monogr. 95:328, 1990. 

Cheskin. L.J., Shabsin HS, Brooner R, Schuster MM, Whitehead WE. Colon motility in opiate addiction 
and naloxone-precipitated withdrawal. Gastrointestinal Motility. 2 (2): 90-5, 1990. 

Fudala, P.J., Heishman S.J., Henningfield J.E., and Johnson R.E. Human pharmacology and abuse 
potential of nalmefene. Clin Pharmacol Ther. (In press). 

Fudala, P.J.. Jaffe J.H., Dax E., and Johnson R.E. Use of buprenorphine in the treatment of opiate 
addiction 1 1 : Physiologic and behavioral effects of daily and alternate-day administration and abrupt 
withdrawal. Clin Pharmacol Ther 1990; 47:525-534. 

Gorelick, DA. "Progression of Dependence in Male Cocaine Addicts" American Journal of Drug and 
Alcohol Abuse (In Press). 

Gorelick, D.A., Irwin MR., Schmidt-Lackner S, and Marder S. Alcoholism among male schizophrenic 
inpatients. Annals of Clinical Psychiatry, 2:19-22, 1990. 

Gorelick. D.A., Paredes A. "Effect of Fluoxetine on Alcohol Consumption in Male Alcoholics." 
Alcoholism: Clinical and Experimental Research (In Press). 

Johnson, RE., Fudala P.J., Fralich J.L. Use of naloxone in the assessment of opiate dependence. Clin 
Pharmacol Ther 1990:47:168 

Johnson, RE., Fudala P.J., Collins C.C., and Jaffe J.H. "Outpatient Maintenance/Detoxification 
Comparison of Methadone and Buprenorphine." NIDA Res. Monogr. 95:389, 1990. 



241 



Kolar, A.F., Brown B.S., Weddington W.W.. and Ball J.C. "A Treatment Crisis: Cocaine Use by Clients 
in Methadone Maintenance Programs,'' Journal of Substance Abuse Treatment 7 (2). (Summer 1990), pp 
101-107. 

Lange, W.R., Fudala P.J.. Dax E.M., and Johnson R.E. Safety and side-effects of buprenorphine in the 
clinical management of heroin addiction. Drug Alcohol Depend 1990; 26:19-28. 

Pickworth. W.B., Lee H., Fudala P.J. Buprenorphine induced pupillary effects in human volunteers. Life 
Sci 1990;47:1269-1277 

Weddington, W.W., Brown B.S.. Haertzen C.A., Cone E.J., Dax E.M., Herning R.I., Michaelson B.S. 
Changes in mood, craving, and sleep during abstinence reported by male cocaine addicts; a controlled, 
residential study. Arch Gen Psychiatry. 1990; 47:861868. 

Weddington. WW., Brown B.S., Haertzen C.A., Hess. J.M., Mahaffey J.R., Kolar A.F., and Jaffe J.H. 
Comparison of amantadine and desipramine combined with psychotherapy for treatment of cocaine 
dependence. Am J Drug Alcohol Abuse. 1991; 17:137-152. 

Weddington, WW. Towards a rehabilitation of methadone maintenance: Integration of relapse 
prevention and aftercare. Int J Addictions. 1991; 25:1205-1228. 

Weddington, WW., Haertzen C.A., Hess J.M., and Brown B.S. Psychological reactions and retention in 
treatment according to HTV-serostatus: a matched-control study. Am J Drug Alcohol Abuse, 1991; 
17:355-368. 

Weinhold, L.L., Funderburk F.R., Summerfelt A.T., and Liebson I.A. Cardiovascular Effects of 
Phenylpropanolamine: A Meta Analytic Examination. Drug Safety, Vol 5 (Suppl. 1), 160-161. 

Weinhold. L.L., and Bigelow G.E. Factors Influencing Assessment of Opioid Miosis in Humans. NIDA 
Research Monograph Series, in press. 

Weinhold, L.L., Jaffe A.B., and Sharpe, L.G. Factors Influencing Self-Administration of Aerosol 
Sufentanil in Rats. NEDA Research Monograph Series, in press. 

Weinhold. L.L. Use of Steroids and Drugs by Athletes. In; Lou Diamont (Ed.) The Psychology of Sports, 
Exercise and Fitness. (In Press). Washington, D.C. Hemisphere Publishing Company. 

Weinhold, L.L., Bigelow G.E. and Preston K.L. (1990) Combination of Naloxone with Buprenorphine in 
humans. NIDA Research Monograph Series: 95, 485. 

Weinhold, L.L., Sharpe, L.G, and Jaffe, J.H. (1990) The Effects of Capsaicin Treatment on Self- 
Administration of Amphetamine Vapor in Rats. NIDA Research Monograph Series: 95, 539. 

PRESENTATIONS & ABSTRACTS 

Ball, J.C "Opening the "Black Box" of Drug Abuse Treatment - Measurement and Evaluation of the 
Treatment Domain," Committee on Problems of Drug Dependence, Inc. (Abstracts), Fifty-Second Annual 
Scientific Meeting, June 10-14, 1990. 

Ball, J.C. "The Status of Methadone Maintenance Treatment in the United States," Australian Methadone 
Conference. Keynote Speaker (Sydney, Australia). July 9-10, 1990. 



242 



Ball. J.C. "Evaluating Methadone Maintenance Programs," Keynote Speaker Midwest Regional 
Methadone Conference. Milwaukee, Wisconsin, September 17-18, 1990. 

Covi, L., Hess J.M. and Haertzen CA. "Why Cocaine and PCP Abusers Seek Treatment." Poster 
presented at the F.S. Key Medical Center Science Day 6/15/90. 

Covi. L.. Baker CD, Hess. J.M. "An Integrated Interpersonal/Cognitive Behavioral Counseling Approach 
to Cocaine Abuse Treatment". Poster presented at the International Conference on Cognitive Therapy 
Philadelphia 10/5/90. 

Cone. E.J., Dickerson SL.. Darwin WD., Fudala PJ., and Johnson RE. Elevated drug saliva levels 
suggest a "depot-like" effect in subjects treated with sublingual buprenorphine. The Committee on 
Problems of Drug Dependence, Inc., (In Press). 

Fudala. PJ.. Johnson RE. Heishman SJ, Cone EJ, and Henningfield JE. A dose run-up and safety 
evaluation of nalmefene HC1 in human volunteers. NIDA Research Monograph 95. 1990:451-452. 

Johnson. RE. Fudala PJ. Jaffe JH. Outpatient comparison of buprenorphine and methadone maintenance 
1. Effects on opiate use and subject reported side effects and withdrawal symptoms. The Committee on 
Problems of Drug Dependence, Inc., (In Press). 

Khan. S, Gorelick, DA, & Nademanee, K: Role of alcohol in cardiac arrhythmias and effect on the 
circadian rhythm of heart rate. Presented at American Society of Addiction Medicine, 21st Annual 
Medical-Scientific Conference, Phoenix, April, 1990. 

Montoya, I. Review of Epidemiological Profiles of Alcohol and Drug Abuse in Latin American 
countries. Bulletin of the Pan American Health Organization, 24: 1990. 

Nademanee, K, Gorelick, DA. Nademanee K. Wilkins JN: Acute and chronic effects of cocaine on 
physiological cardiovascular parameters in men. Presented at American Society of Addiction Medicine, 
21st Annual Medical-Scientific Conference, Phoenix, April, 1990. 

Nolimal, D. Drinking, Suicide and Maintenance of Ethnic Identity among the Slovenes. Surveyor, 23: 
28, 1990. 

Tashkin DP. Gorelick, DA, Simmons. MF, Khalsa, ME Chang, P. Coulson, AH, & Gong, H, Jr: 
Respiratory effects of cocaine freebasing among habitual users. Presented at American Society of 
Addiction Medicine, 21st Annual Medical-Scientific Conference, Phoenix, April, 1990. 

Tashkin. DP, Gorelick, DA, Simmons MF, Khalsa ME, Chang P, Coulson AH, & Gong H, Jr: Respiratory 
symptoms and lung function in heavy habitual "crack" smokers. Presented at World Conference on Lung 
Health, International Union Against Tuberculosis and Lung Disease/American Lung 
Association/American Thoracic Society, Boston, May, 1990. (American Review of Respiratory Disease 
14LA776, 1990). 

Weddington WW. Brown BS, Haertzen CA, Hess JM, Mahaffey JR, Kolar AF, Jaffe JH. Amantadine and 
desipramine for treatment of cocaine dependence. Nat Inst Dr Ab Monar Ser 95 (ADM) 90-1663, pp. 
483-484, 1990. 

Weddington WW, Kolar AF, Brown BS, Ball JC: A treatment crisis: Cocaine use by clients in methadone 
maintenance programs. Nat Inst Dr Ab Monar Ser (ADM) 91-1753, pp. 365-366, 1991. 



243 



Weddington WW, Brown BS. Cone EJ. Haertzen CA, Dax EM, Herning RI. Michaelson BS. Changes in 
mood, craving and sleep during acute abstinence reported by male cocaine addicts. Nat Inst Dr Ab 
Monoar Ser (ADM) 91-1753, pp. 453-454, 1991. 

Wilkins, JN & Gorelick, DA: Neuroedocrine effects of alcohol and alcohol withdrawal. Presented at 
American Society of Addiction Medicine. 21st Annual Medical-Scientific Conference, Phoenix, April, 
1990. 

Wilkins JN & Gorelick. DA: PCP use and alcoholism in an urban VA psychiatric population. Presented at 
conference on "Aging in the 1990's: Alcohol and Other Drug Abuse" sponsored by Western Michigan 
University, Novi, Michigan, April. 1990. 

Wilkins JN, Shaner AL, Patterson CM. Setoda D, and Gorelick, DA: Screening drug evaluation subjects 
for substance abuse: descrepancies between patient report, clinical assessment and urine analysis. 
Presented at New Clinical Drug Evaluation Unit Program, Key Biscayne, FL, June, 1990. 

Wong, M & Gorelick, DA: Effect of alcohol on ventricular function in male alcoholics. Presented at 
American Society of Addiction Medicine 21st Annual Medical-Scientific Conference. Phoenix, April, 
1990. 



244 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00139-01 PTL 

Title of Project: Effects of Carbamazepine on Cocaine Self-Administration 

Principal Investigators: David A. Gorelick, M.D., Ph.D., Linda Weinhold. Ph.D., Frances Rudnick 
Levin, M.D., Jack Henningfield, Ph.D., Ed Cone, Ph.D., David Newlin, Ph.D., Ronald Herning, Ph.D., 
W.RLange, M.D. 

Cooperating Units: Chemistry and Drug Metabolism Lab, Vulnerability Lab, Office of Medical Affairs. 

Lab/Branch: Pharmacotherapy Lab/Treatment Branch 

Section: NONE 

Addiction Research Center. National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 4.0 Professional: 1.0 Other: 3.0 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

This double-blind, double dummy, placebo-controlled (diphenhydramine-25 mg BID) residential study 
assesses the psychological, physiological, cardiovascular, cognitive, electroencephalographic, and 
pharmacokinetic effects of the carbamazepine-cocaine interaction in cocaine-abusing subjects not 
currently dependent on other drugs. 

Eighteen subjects are randomly assigned to 4 parallel groups: (1) low plasma carbamazepine levels (1-3 
mg/L), (2) placebo short-stay (6 weeks); (3) middle plasma carbamazepine levels (4-7 mg/L); and (4) 
placebo long-stay (9 weeks). Subjects can self-administer cocaine-25 mg IV (or blank) or receive a 
monetary reward up to thrice daily 3 days each week by making a stimulus-controlled operant response. 
On self-administration days, subjects undergo 24-hour ambulatory monitoring of cardiovascular function, 
and answer computer administered questions on their subjective state. Subjects' response to cocaine- 
associated stimuli and their EEG and cognitive function are assessed periodically. Thus, the influence of 
carbamazepine on cocaine reinforcement and cocaine- induced psychological and physiological effects 
can be assessed. Blood, saliva, and hair samples are collected periodically to assess cocaine and 
carbamazepine pharmacokinetics. 

This is the largest, most detailed study to date investigating the clinical pharmacology of the 
carbamazepine-cocaine interaction. 



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NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00100-01 CTL 

Title of Project: Comparison of Detoxification Treatment for Opiate Addiction 

Principal Investigators: Lawrence J. Cheskin. M.D.. Paul Fudala, Ph.D. 

Cooperating Units: Neuropharmacology Lab, Vulnerability Lab 

Lab/Branch: Pharmacotherapy Lab/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 

21224 



Total Man Years: 1.2 



Professional: .2 



Other: 1.0 



Check Appropriate Boxes: 



X Human Subjects 

Minors 

Interviews 



Human Tissues 



Neither 



Summary of Work 

This study is a double-blind, double-dummy comparison of buprenorphine (1-4 mg si daily), a partial 
opiate agonist, vs. clonidine (0.2 - 0.9 mg po daily), an alpha-2 adrenergic agonist, in the treatment of 
acute opiate withdrawal. Sixteen patients are randomly assigned to receive one of the 2 medications for 3 
days, followed by 14 days of inactive placebo. Patients are also stratified by degree of opiate dependence, 
as measured by a cutoff opiate withdrawal score following naloxone challenge (0.4 mg im). 

Outcome measures include both physiological and psychological signs and symptoms of opiate 
withdrawal. In addition, optional subprotocols involve measuring vagal and sympathetic tone as reflected 
in variations in cardiac interbeat interval (in collaboration with the ARC Vulnerability Lab), and cerebral 
metabolism using positron emission tomography (PET) with 2-F-DG (in collaboration with ARC 
Neuropharmacology Lab and Hopkins PET Center). 

To date, 1 2 patients have completed the study, which is the first direct, controlled clinical comparison of 
buprenorplune with a standard treatment for acute opiate withdrawal. 



246 



NOTICE OF INTRAMURAL RESEARCH Z01 DA 00101-01 PTL 

Title of Project: HIV Infection. High-Risk Behaviors, and Drug Abuse Treatment 

Principal Investigators: David A. Gorelick, M.D., Ph.D., Ivan D. Montoya, M.D., MPH, W.R. Lange, 
M.D., Judy Hess, M.A., Weddington, M.D. 

Cooperating Units: Office of Clinical Director 

Lab/Branch: Pharmacotherapy Lab/Treatment Branch 

Section:NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 

21224 

Total Man Years: 0.50 Professional: 0.25 Other: 0.25 
Check Appropriate Boxes: 



X Human Subjects Human Tissues Neither 

Minors 

X Interviews 



Summary of Work 

This two-phase project evaluates the relationship between sociodemographic, psychosocial, cognitive, 
and drug use variables and HIV infection in drug abusers, and the influence of HTV infection on drug 
abuse treatment compliance and outcome. The first phase involves retrospective analysis of data already 
collected on ARC subjects, using two analytic approaches: I ) case-control comparisons of subjects HIV 
antibody positive or negative who are matched on appropriate variables and 2) multivariate analyses 
using the entire sample of subjects to determine variables associated with HIV infection and treatment 
compliance and outcome. The second phase prospectively collects data on HTV infection high-risk 
behaviors and attitudes using self-report questionnaires and structured interviews. Data is collected from 
ARC treatment research subjects at treatment entry and again at follow-up in order to evaluate possible 
changes due to drug abuse treatment or drug abstinence. 

Initial analysis of retrospective case-control data from 22 HIV-antibody positive, physically 
asymptomatic cocaine addicts and 22 matched HTV seronegative cocaine addicts indicated that 
seropositive subjects were significantly more likely to be HTV cocaine users. Otherwise, there were no 
significant differences between the two groups in sociodemographic, psychosocial, cognitive, or 
treatment compliance variables. 



247 



Publications: 

Weddington, WW. Haertzen CA, Hess JM, Brown BS: Psychological reactions and retention in 
treatment according to HTV-serostatus: a matched-control study. Am J Drug Alcohol Abuse, 1991: 
17:355-368. 



248 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00107-01 PTL 

Title of Project: Esterase activity in human cocaine abusers 

Principal Investigators: David Gorelick, M.D., Ph.D., Linda Weinhold, Ph.D., Raymond Woosley, 
M.D. 

Cooperating Units: Department of Pharmacology, Georgetown University 

Lab/Branch: Pharmacotherapy Lab/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 0.35 Professional: 0.1 Other: 0.25 

Check Appropriate Boxes: 

x Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

Esterase enzymes in blood are die major metabolic pathway for cocaine in humans. There is population 
variability in enzyme activity, some of which is genetically based, yet no published studies to date have 
evaluated enzyme activity in cocaine abusers or the correlation between enzyme activity and response to 
cocaine. This study, done in collaboration with Dr. Raymond Woosley, Department of Pharmacology, 
Georgetown University, measures plasma and RBC esterase activity in cocaine abusing research subjects 
at the ARC to determine the influence of this metabohc parameter on response to cocaine and on 
treatment outcome. 



249 



2. Clinical Trials Laboratory - Paul Fudala, Ph.D., Chief 



Overview 



The goal of the Clinical Trials Laboratory is to test the safety and efficacy of new treatments for drug 
abuse, including both pharmacologic and non-pharmacologic (psychosocial) treatments and their 
interactions. Studies are conducted in a realistic community (outpatient) setting to gain clues to the 
effectiveness, safety, and mechanism of action of new treatments that might lead to more intensive study, 
and to assess various environmental factors and patient characteristics (e.g., availability of illicit drugs, 
personality traits) affect compliance, treatment outcome, duration in treatment, and other clinically 
relevant variables. Long-term follow-up is conducted to assess the duration of beneficial, as well as 
potentially adverse, effects of treatment. 



Summary of Research 

A. Efficacy of fluoxetine and desipramine in the treatment of cocaine and PCP dependence. 

The first phase of this outpatient study assessed the efficacy of fluoxetine, a serotonergic reuptake 
blocker, for the treatment of cocaine dependence. Fifty-three subjects were admitted to the study; 
analyzable data was obtained for 46 of these. All subjects received counseling twice weekly. No 
significant differences were found between treatment groups (fluoxetine 20, 40, 60 mg, or active placebo) 
for any dependent variable (e.g., cocaine-positive urine, subject-reported craving for cocaine), regardless 
of whether subjects were grouped by fluoxetine dosage or plasma levels. The second phase of this study 
involved comparisons between fluoxetine and placebo in PCP abusers and desipramine and placebo in 
cocaine abusers. Forty-seven subjects were enrolled into the desipramine protocol. Subjects received 
either desipramine. 300 mg per day or inactive placebo for 12 weeks. All subjects received counseling 
twice weekly. Forty-four subjects were enrolled into the fluoxetine protocol. Subjects received either 
fluoxetine, 20 mg per day or active placebo (diphenhydramine 12.5 per day) f or 12 weeks. All subjects 
received counseling once weekly. Data analysis is currently ongoing. 

B. Buprenorphine/ methadone comparison - maintenance and detoxification 

The purpose of this study was to determine the effectiveness of buprenorphine in maintaining opiate- 
dependent individuals in non-residential treatment as compared to the prototypic treatment drug 
methadone. Additional information regarding the medical safety of the two treatments, as well as 
pharmacokinetic data, were obtained from analyses of blood and urine. Preliminary analyses of the data 
have indicated that: 1 ) no differences were observed between treatment groups for self-reported opiate 
withdrawal symptoms, 2) no pattern of results was observed between groups with respect to subject- 
reported adverse effects which could be related to the study medications, 3) the percentage of missed 
clinic visits did not differ significanfly between treatment groups, 4) subject retention rates at the 
completion of the maintenance phase (study day 119) were approximately double for the buprenorphine 
and the higher-dose methadone treatment groups compared to the lower-dose methadone group, 5) 
buprenorphine treatment was associated with more urine samples negative for opiates compared to either 
methadone group, and 6) no significant or consistent differences were observed between groups for 
cocaine-positive urine samples. It is anticipated that this will be considered a pivotal study by the FDA. 
potentially leading to the approval of buprenorphine as a pharmacotherapy for opiate dependence. 

C. Assessment of nalmefene glucuronide as a selective antagonist of gut opioid action 

A single-blind ascending dose study was conducted to determine whether nalmefene glucuronide 



250 



precipitates withdrawal symptomatology in opiate-(methadone-) dependent individuals. If the 
glucuronide were shown to exert a selective effect on the enteric nervous system without antagonizing 
central opiate actioas, it could have utility for patients who require high doses of opiates but who suffer 
from gastrointestinal side effects (e.g., constipation). However, withdrawal symptoms were induced at 
relatively low doses (e.g., 1 mg po daily). Thus, nalmefene glucuronide does not appear to be useful for 
constipation associated with the administration of exogenous opiates. 

D. Impact of differing intensities of drug abuse counseling 

While much research attention has focused on pharmacologic treatment for drug abuse, few studies have 
addressed the efficacy of psychosocial treatments such as counseling. This 12-week outpatient study is 
evaluating the effectiveness of individual counseling as a treatment for cocaine dependence. Patients are 
randomly assigned to one of three counseling frequencies: twice weekly, once weekly, or every other 
week. All counseling is delivered according to a specified therapy manual in which all counselors have 
been trained, thus assuring that all patients receive the same type of treatment. Treatment outcome 
measures include urine toxicology, subject-reported drug use, cocaine craving, depressive symptoms, and 
psychosocial functioning. 



E. A double-blind comparison of carbamazepine and placebo for the treatment of cocaine abuse 

The anti-con vulsant medication carbamazepine has been suggested as a treatment for cocaine abuse, but 
there are as yet no published double-blind, placebo-controlled treatment studies establishing its efficacy. 
This eight-week outpatient study uses such a design to evaluate the efficacy and safety of carbamazepine 
(up to 800 mg daily) plus individual counseling in the treatment of cocaine dependence. Treatment 
outcome measures include urine toxicology, subject-reported drug use, cocaine craving, depressive 
symptoms, and psychosocial functioning. 

F. Reasons for seeking drug abuse treatment 

This study is collecting data on the self-reported reasons why drug abusers seek treatment. Data will be 
analyzed in terms of differences among drug abusing populations and for possible predictive factors for 
treatment compliance and outcome. 



3. Drug Abuse Treatment Evaluation Unit - John Ball, Ph.D., Chief 

The Treatment Evaluation Unit applies a comprehensive schema for evaluating existing drug abuse 
treatment, assessing 89 variables in four areas: patient history and characteristics, program 
characteristics, treatment services provided, and treatment outcome. The goal is to determine the efficacy 
of existing treatment programs and identify variables associated with successful outcome. 

Dr. Ball has applied his methodology to the evaluation of methadone maintenance treatment for opiate 
dependence, with data analysis largely completed The next study planned will evaluate existing drug 
abuse treatment programs in the Baltimore area, including those that treat cocaine users. 



251 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00090-02 CTL 

Title of Project: Buprenorphine/Methadone Comparison - Maintenance and Detoxification 

Principal Investigators: RE. Johnson. Pharm.D.. P.J. Fudala, Ph.D., W.R. Lange, M.D. 

Cooperating Units:NONE 

Lab/Branch: Clinical Trials Lab/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 12.3 Professional: 3.3 Other: 9.0 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 



Summary of Work 

Previous residential studies conducted at the ARC showed that heroin dependent individuals may be 
rapidly inducted onto buprenorphine without producing clinically significant opiate-withdrawal 
symptoms, may be maintained on daily or alternate-day buprenorphine dosing schedules, and experience 
a mild to moderate withdrawal syndrome after abrupt withdrawal of buprenorphine. Results from dose- 
ranging studies indicated an appropriate dose for use in non-residential maintenance treatment. 

The purpose of this study was to determine the effectiveness of buprenorphine (8 mg si daily) in 
maintaining opiate-dependent individuals over 6 months of non-residential treatment as compared to the 
prototypic treatment drug methadone (20 mg or 60 mg daily). 162 opiate-dependent subjects were 
randomly assigned after stratification by age, sex, and the results of a naloxone challenge. 

This study, the largest clinical trial to date assessing the effectiveness of buprenorphine for the treatment 
of opiate dependence, found buprenorphine as effective as methadone-60 mg on almost all outcome 
measures, and both significandy more effective than methadone-20 mg. There was no significant 
difference among groups in urine samples positive for cocaine. 



252 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00081-03 CTL 

Title of Project: Double-Blind Comparison of Desipramine. Fluoxetine and Bromocriptine for the 
Treatment of Cocaine and PCP Abuse 

Principal Investigators: L. Covi, M.D.. C. BakerJvl.A.. J.M. Hess, M.A. 

Cooperating Units: Office Of Medical Affairs 

Lab/Branch: Clinical Trials Lab/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse. Baltimore, MD 

21224 

Total Man Years: 2.0 Professional: 0.5 Other: 1.5 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 

Summary of Work 

This series of 3 double-blind, placebo-controlled (active placebo = diphenhydramine - 12.5 mg) studies 
examined the efficacy of medication plus individual counseling (Rounsaville's adaptation of interpersonal 
psychotherapy) for 12 weeks as outpatient treatment of drug abuse. Outcome was assessed by drug use 
(both urine toxicology and self-report), drug craving, mood, and psychosocial functioning (Addiction 
Severity Index). All subjects were followed up 3, 6, 12 months after active treatment. 

In the first study, 53 cocaine abusers (ages 21-60) were randomly assigned to receive either fluoxetine- 20 
mg <n=U), 40 mg (14), 60 mg (12), or placebo (16) daily. All patients also received counseling twice 
weekly. There were no significant outcome differences among groups, whether analyzed in terms of 
medication dose or achieved plasma level (> vs < 100 ng/ml). At follow-up, all groups showed some 
improvement, with no group differences. 

In the second study, 45 PCP abusers were randomly assigned to receive either fluoxetine-20 mg daily or 
placebo, plus counseling twice weekly. In the third study, 46 cocaine abusers were randomly assigned to 
receive either desipramine- 300 mg daily or placebo, plus counseling once weekly. Data from both studies 
are being analyzed. 



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NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00091-01 CTL 

Title of Project: Assessment of Nalmefene Glucuronide as a Selective Antagonist of Gut Opioid Action 
Principal Investigators: Lawrence J. Cheskin, M.D., R.E. Johnson, Pharm.D. 
Cooperating Units: NONE 
Lab/Branch: Treatment 
Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore.MD 

21224 

Total Man Years: 0.5 Professional: 0.2 Other: 0.3 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 



Summary of Work 

A single-blind ascending dose study was conducted in 5 subjects to determine whether nalmefene 
glucuronide precipitates withdrawal symptomatology in opiate-( methadone) dependent individuals. If the 
glucuronide were shown to exert a selective effect on the enteric nervous system without antagonizing 
central opiate actions, it could be useful in patients requiring high doses of opiates, but who suffer from 
gastrointestinal side effects (e.g., constipation). However, withdrawal symptoms were induced at 
relatively low doses (e.g., 1 mg po daily). Thus nalmefene glucuronide does not appear to be useful for 
constipation associated with the administration of exogenous opiates in opiate-dependent subjects. It may 
be useful, however, in patients with constipation who are not opiate-dependent, such as persons suffering 
from constipation - predominant irritable bowel syndrome. 



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PUBLICATIONS: 

Fudala P.J.,Ph.D., Heishman S.J., Ph.D., Henningfield J.E.Ph.D., and Johnson R.E., PharmD. Human 
Pharmacology and Abuse Potential of Nalmefene. Clin. Pharm. and Ther. Vol. 49, No. 3, pp. 300-306, 
March, 1991. 



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NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00508-01 CTL 

Title of Project: Reasons for seeking drug abuse treatment 

Principal Investigators: Lino Covi, M.D., Judy Hess, M.A., C. Haertzen, Ph.D. 

Cooperating Units: Clinical Pharmacology Branch 

Lab/Branch: Clinical Trials/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 

21224 

Total Man Years: 0.3 Professional: 0.1 Other 0.2 

Check Appropriate Boxes: 



X Human Subjects Human Tissues Neither 

Minors 

Interviews 



Summary of Work 

In order to identify factors possibly influencing treatment compliance and retention, 206 applicants for 
outpatient pharmacological treatment of cocaine or PCP dependence gave self-reported reasons for 
seeking treatment, using a new questionnaire developed by the P.I. There were significant differences in 
reasons given between cocaine vs PCP abusers and males vs females. Subjects not motivated by the cost 
of their drug abuse stayed in treatment twice as long (6 weeks) as those who were motivated by cost. 
Currently, data are being collected to compare counselors' ratings of treatment motivations with the self- 
report instrument. 



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Publication 

L. Covi. J.M Hess, CA Haertsen: Why Cocaine and PCP Abusers Seek Treatment. Poster presented at the 
Science Day - Francis Scott Key Medical Center. May, 1990. 



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NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00098-02 CTL 

Title of Project: Impact of Differing Intensities of Drug Abuse Counseling 

Principal Investigators: L. Covi, M.D., C.Baker, M.A., J. M.Hess, M.A. 

Cooperating Units: Office of Medical Affairs 

Lab/Branch: Clinical Trials Lab/Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.0 Professional: 0.5 Other: 1.5 

Check Appropriate Boxes: 

x Human Subjects Human Tissues Neither 

Minors 

Interviews 



Summary of Work 

This study evaluates the influence of frequency of individual counseling on the effectiveness of cocaine 
abuse treatment. Subjects meeting DSM-1I1-R criteria for cocaine dependence will be randomly assigned 
(50 per group) to one of three treatment conditions plus urine monitoring for 12 weeks: a) twice weekly 
counseling, b) once weekly counseling, or c) counseling every other week. 

All counseling is delivered according to a specified therapy manual integrating aspects of interpersonal, 
cognitive, and behavioral approaches to drug abuse counseling. In the pilot phase, 27 subjects were 
treated to refine the manual and train counselors in the therapy. 

Treatment outcome will be assessed by urine toxicology twice weekly (collected under direct staff 
observation) , breath alcohol analysis, self-reported drug use and cocaine craving, psychological 
symptoms, and psychosocial functioning (Addiction Severity Index). Follow-up at 3, 6, and 12 months 
will assess any long-term effects of treatment. 

To date, 25 subjects have entered the study, of whom 6 received initial evaluation only, with no actual 
treatment. 



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Publication: 

L. Covi, M.D., CD. Baker, M.A., and J.M. Hess, M.A.: "An Integrated Interpersonal/Cognitive 
BehavioraJ Counseling Approach to Cocaine Abuse Treatment". Poster presented at the International 
Conference on Cognitive Therapy, Philadelphia 10/5/90. 



259 



NOTICE OF INTRAMURAL RESEARCH PROJECT Z01 DA 00106-01 CTL 

Title of Project: Evaluation of methadone maintenance treatment for opiate dependence 

Principal Investigators: John Ball, Ph.D. 

Lab/Branch: Treatment Branch 

Section: NONE 

Addiction Research Center, National Institute on Drug Abuse, Baltimore, MD 21224 

Total Man Years: 2.0 Professional: 1.0 Other: 1.0 

Check Appropriate Boxes: 

X Human Subjects Human Tissues Neither 

Minors 

Interviews 



Summary of Work 

This project applies a comprehensive schema for evaluating existing drug abuse treatment, assessing 89 
variables in four areas: patient history and characteristics, program characteristics, treatment services 
provided, and treatment outcome. The goal is to determine the efficacy of existing treatment programs 
and identify variables associated with successful outcome. 

Dr. Ball has applied this methodology to the evaluation of 6 methadone maintenance treatment programs 
for opiate dependence, with data analysis largely completed 

The findings from this large-scale, prospective, systemic treatment outcome study indicate that 
methadone maintenance is effective in reducing drug use, criminal activity and some HIV infection high- 
risk activities, but that treatment effectiveness can vary widely among programs, depending on several 
variables such as methadone dose, staff qualifications, and staff turnover. 



260 



Publications: 

Ball. John C. "A Comprehensive Evaluation of Methadone Maintenance Programs in New York City, 
Philadelphia and Baltimore," Advances in Alcohol & Substance Abuse (Guest Issue), (In press). 

Ball. John C. "A Schema for Evaluating Methadone Maintenance Programs." In: L.S. Harris (ed.), 
NJDA Research Monograph: Proceedings of the 51st Annual Scientific Meeting of the Committee on 
Problems of Drug Dependence. 1989 (In press). 

Ball, John C. "The Effectiveness of Methadone Maintenance Treatment in the United States - An 
overview." Presented at the "What Works Conference" in New York, October 22-24, 1989. (In press). 

Kolar A.F.. Ball, John C, Brown F.S.. and Weddington WW. "A Treatment Crisis: Cocaine Use by 
Clients in Methadone Maintenance Programs," Journal of Substance Abuse Treatment 7 (2), (Summer 
1990), pp 101-107. 



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