George F. Koob - US grants

Arginine vasopressin (AVP), a posterior pituitary peptide hormone, has been shown to have several non-renal actions including the maintenance of learned avoidance behaviors, and recently, in humans, improvement in cognitive functioning. Preliminary research in our laboratory has confirmed some of these behavioral effects in animals and begun an attempt to examine the behavioral and physiological basis of this effect. The present proposal for research will focus on determining the physiological mechanism by which AVP produces its behavioral effects. Behavioral experiments will concentrate on examining the effects of AVP in rats on two tests of "memory", an active avoidance task and a spatial forgetting test, and one test of "emotionality" the Geller-Seifter conflict test. These tests have been chosen to control for the nature of the unconditioned stimulus (aversive vs. appetitive), motor debilitating effects, and motivational effects of AVP itself. The importance of peripheral mechanisms related to the classical endocrinological effects of AVP will be determined by using these behavioral tests and analogs of AVP, some of which are specific antagonists of certain endocrinological effects. In addition, an attempt will be made to identify the distribution and site of action of exogenously applied AVP using radioimmunoassay and intracerebral injections. This basic research directed at understanding the mechanism of action of the effects of vasopressin on behavior has important implications for the possible therapeutic use of vasopressin in cognitive disorders associated with aging, and other biologically based learning disorders.

This proposal will continue to characterize the behavioral effects of arginine vasopressin (AVP) and to explore its mechanism and site of action. Based on studies with AVP analogs, one of which is a vasopressor AVP antagonist peptide, our hypothesis is that peripherally derived (pituitary) AVP alters behavior secondary to its peripheral, physiological effects, but that central AVP has behavioral actions that are separate and of unknown mechanism. An interoceptive stimulus (osmotic challenge with hypertonic saline), known to release AVP, produced behavioral effects similar to administration of exogenous AVP and these behavioral effects were reversed by systemic administration of the AVP antagonist peptide. Studies are designed to characterize further these effects and to examine the site of action for the behavioral actions of hypertonic saline by using central and peripheral administration of less lipophilic AVP antagonists. Further studies are designed to explore the neural site of action for the behavioral effects of central nervous systems (CNS) AVP by using local intracerebral injection of AVP and AVP antagonists. Behavioral tests will include both aversively and appetitively motivated tasks for measuring learning and memory such as active and inhibitory avoidance, a water finding task and a social memory task. Studies also have been designed to explore the functional role of extrahypothalamic and hypothalamic-pituitary AVP by stimulation of AVP cell bodies in the paraventricular nucleus, bed nucleus of the stria terminalis and amygdala and measuring the effects on release of AVP using push-pull cannulation of AVP terminal regions. Subsequently stimulation effective in releasing endogenous central or pituitary AVP will be used to mimic the effects of AVP on behavior. Oxytocin, the second neurohypophyseal nonapeptide, has been reported to have behavioral actions opposite to that of AVP. We propose to extend these earlier observations by characterizing the behavioral effects of both systemically and centrally administered oxytocin and by exploring its mechanism and site of action using oxytocin antagonist peptides. This basic research directed at understanding the mechanism of action for the effects of AVP and oxytocin has important implications, not only for our understanding of the organization of CNS-pituitary interactions in behavioral responses to homeostatic challenge, but also for our understanding of the biological basis of memory and learning.

This Project will extend ongoing recent work in our laboratory where we established the region of the nucleus accumbens as an important substrate for the reinforcing properties of heroin in non-physically dependent rats self-administering heroin intravenously. The present proposal will continue these studies by exploring further the central mechanisms involved in opiate reinforcement and will extend these studies to exploring the central mechanisms involved in opiate dependence. The experiments proposed are designed to examine the role of the nucleus accumbens and its efferent projections in opiate reinforcement in non dependent animals, and to compare these results with similar studies in physically dependent animals. At the same time the neuronal sites of action important for precipitated opiate withdrawal will be explored in dependent animals by using local intracerebral injections of methylnaloxonium, a selective opiate antagonist with greatly reduced lipophilicity and diffusability. Withdrawal measures will include both unconditioned and conditioned behavior. Significant evidence exists to show that rats, monkeys and man show behavioral and physiological manifestation of narcotic withdrawal as a response to previously neutral stimuli that have been paired with a state of precipitated abstinence. An attempt will be made to define the functional substrate for the phenomenon of conditioned narcotic withdrawal by attempting to reverse conditioned withdrawal with local intracerebral injection of morphine or opioid peptides. These studies have important implications for our understanding of the neural substrates involved in opiate seeking behavior and the addictive process.

DESCRIPTION (provided by applicant): Drug addiction in humans is characterized by compulsive drug taking and has been conceptualized as an increased motivation to seek drugs. Still largely unknown are what neurochemical/neurocircuitry changes occur that provide the motivational basis for vulnerability to escalation in drug intake over time. During the previous funding period, an animal model of escalation of cocaine intake was established using prolonged access to cocaine that provides heuristic value for exploring the neurobiological changes that may lead to vulnerability to escalation in cocaine intake and relapse. Neuropharmacological studies during the previous funding period have pointed to changes in reward neurotransmitter neurocircuitry (dopamine and opioid peptides) and changes in brain stress systems as contributing to the escalation process. The purpose of the present proposal is to systematically explore the neurobiological mechanisms responsible for expression of cocaine escalation. The overall hypothesis under test is that sufficient exposure to cocaine leads to changes in specific elements of the neurocircuitry of the extended amygdala to produce progressive elevation in drug intake. A subhypothesis is that the neurobiological basis of this transition to drug escalation reflects compromised dopamine and opioid peptide function and a recruitment of the brain corticotropin-releasing factor (CRF) and norepinephrine systems in different elements of the extended amygdala. The purpose of the proposed studies is to explore the role of specific dopamine receptor subtypes (Specific Aim 1), the role of opioid peptides and specific opioid receptor subtypes (Specific Aim 2), the role of the CRF systems and specific CRF receptor subtypes (Specific Aim 3), and the role of the norepinephrine systems and specific norepinephrine receptor subtypes (Specific Aim 4) in the expression of cocaine escalation in specific subregions of the extended amygdala. These experiments will provide critical information about the brain changes that accompany the transition from stable, limited-access cocaine intake to prolonged access, escalated cocaine intake. Such information is critical for the development of new treatments for cocaine addiction as well as for the development of strategies to identify individuals vulnerable for the development of cocaine addiction.

This award will support collaborative research in neurobiology between Dr. George Koob, Scripps Clinical and Research Foundation, La Jolla, California, and Professor Michel LeMoal, University of Bordeaux II, France. The objective of the proposed research is to study the role of the peptide neurotransmitter corticotronin releasing factor (CRF) as a primary mediator of the behavioral state of stress and the behavioral responses to stress in the central nervous system. Recent studies have shown that CRF liberated directly into the central nervous system may have neurotropic actions important for mobilizing behavioral responses to stress. The investigators have designed a set of experiments to identify the specific neural substrates for these behavioral actions by direct local interacerebral injection of CRF and CRF antagonists. In addition experiments are proposed to explore the possible mediation of these CRF effects via the central and peripheral noradrenergic systems. An attempt will be made to modify behavioral responses to CRF using specific pharmacological and neurotoxin induced inactivation of the central and peripheral noradrenergic systems. These studies have important implications for the role of pituitary peptides in brain function, and may provide important clues to the pathophysiology associated with behavioral disorders as diverse as depression, anxiety, and obesity. Over the past ten years Dr. Le Moal and his research group have developed and explored systematically behavioral and endocrine correlates of stress, coping and behavior. Dr. Koob's expertise is in classic behavioral pharmacology and basic neurobiology. The expected benefits to U.S. science from this project include not only the exchange of special behavioral techniques, but also the gain of valuable expertise regarding the measurement of brain function via behavioral techniques.

This is a competing renewal application to continue to study the neural mechanisms of opiate reinforcement and dependence. Work during the previous funding period has established separate neural sites involved in the classic physical and motivational signs of opiate withdrawal. The region of the locus coeruleus and other brain sites such as the hypothalamus appear to have an important role in the expression of physical signs such as wet dog shakes, ptosis and jumping. However, the region of the nucleus accumbens and amygdala appear to be important sites for the motivational effects of opiate withdrawal as measured by lever response suppression and place aversions and are the sites least involved in the physical signs of withdrawal. These same sites are also responsible for the reinforcing effects of opiates in both the non-dependent and dependent state supporting the hypothesis that elements in this limbic- extrapyramidal interface are the critical determinants of the motivational components of opiate dependence. The purpose of the present proposal is to continue to test this hypothesis by further exploring the neural basis for both the positive and negative reinforcing effect of opiate dependence. Local intracerebral microinjections of the hydrophilic opiate antagonist methylnaloxonium will be used to explore the subregions of the extended amygdala in opiate reinforcement as measured by intravenous self- administration (Specific Aim 1) and the motivational effects of opiate withdrawal as measured by response suppression, place aversion and intracranial self-stimulation (Specific Aim 2). An animal model for the positive conditioned effects of opiates in non-dependent and dependent rats will -be developed using operant schedules of heroin self- administration (Specific Aim 3). The motivational effects of both the positive and negative conditioned reinforcing stimuli will be assessed in both non-dependent and dependent rats using intravenous self- administration of heroin (Specific Aim 4). Finally, the neural substrates for the positive and negative conditioned reinforcing stimuli established by pairings with opiates or opiate withdrawal, respectively, will be explored using cell body specific neurotoxic lesions with focus on amygdala- accumbens interactions (Specific Aim 5). These studies will go far towards elucidating the neural mechanisms mediating the brain changes critical for the development of opiate dependence and the neural- mechanisms responsible for the association of this dependence with previously neutral stimuli. Knowledge of these neural substrate will provide information critical for the prevention, treatment and medication of opiate abuse and drug abuse in general.

This proposal requests equipment necessary to extend preliminary studies combining intracerebral dialysis with behavioral measurement in freely moving rats during intravenous self-administration of cocaine. To that end facilities for in vivo microdialysis supported by an appropriate microsampling system will be established that will permit frequent sampling of synaptic concentrations of dopamine and its metabolites during 3 hour self-administration sessions. the purpose of these experiments will be to study the effects of self-administered cocaine on dopamine release in several terminal areas that have been implicated in cocaine reward and in the development of dependence and psychopathology associated with cocaine abuse. Synaptic levels of dopamine will be monitored at 5 minute intervals in the nucleus accumbens, prefrontal cortex and corpus striatum and compared to the animal's pattern of responding for the drug. Attempts will also be made to investigate specific changes in dopamine function after repeated as opposed to acute cocaine self administration. These studies have important implications for an understanding of critical circuitries in psychostimulant reward and dependence.

Description: (Taken directly from the application). Work during the corticotropin-releasing factor (CRF)antagonist, alpha helical CRF, has releasing factor (CRF) antagonist, alpha helical CRF, has provided has a role in behavioral responses to stress that is independent of its role in behavioral responses to stress that is independent of its role intracerebroventricularly (ICV) reverses the behavioral suppression intracerebroventricularly (ICV) reverses the behavioral suppression stress, and the behavioral suppression associated with the non-physical stress and the behavioral suppression associated with the non-physical ICV antagonist appears to be independent of the nature of the stressor and of ICV antagonist appears to be independent of the nature of the stressor The antagonist injected intracerebroventricularly reverses a number of stressor. The antagonist injected intracerebroventricularly reverses a maze, stress-induced decreases in food intake, increases in stress-induced elevated plus maze stress-induced decreases in food intake increases in the CRF antagonist have identified the central nucleus of the amygdala and intracerebral injection of the CRF antagonist have identified the the bed nucleus of the stria terminalis as specific brain sites involved paraventricular nucleus of the hypothalamus and the bed nucleus of the potent and longer acting CRF antagonist, CRF 12-431, has been identified. actions of the CRF antagonist. In addition a more potent and longer identifying the specific brain sites responsible for the anti-stress present proposal is to extend these observations by identifying the brain sites may be involved in different behavioral responses to either antagonists. The hypothesis under test is that different brain sites may the basal forebrain (amygdala and bed nucleus of the stria terminalis), the physical stressors. The focus will be on brain sites of the basal (physical) and social defeat (non-physical) stressors. Dependent measures hypothalamus and the locus coeruleus of the pons in the context of swim on the elevated plus maze and decreases in food intake) and stress-induced measures will focus on stress induced suppression of behavior (decreased will be directed at testing the hypothesis that brain CRF is involved in stress-induced increases in behavior (fear potentiated startle). In including cocaine, opiates, nicotine and benzodiazepines. Finally, CRF is involved in the stress response associated with withdrawal from duration of action for the anti-stress actions of new CRF antagonists and Finally significant effort will be directed at exploring the efficacy, with a focus on testing the hypothesis of potential CRF receptor subtypes. antagonists and at testing functional deficits of transgenic CRF for our understanding of the role of CRF in behavioral responses to stress potential CRF receptor subtypes. The results generated from these including anxiety and drug abuse. role of CRF in behavioral responses to stress and may provide insight into the role of the brain CRF in psychopathology including anxiety and drug abuse.

Ethanol has reinforcing properties in man that are thought to contribute to alcohol abuse and alcoholism. In rats ethanol is readily self-administered orally when taste factors are minimized by preexposing the rats to sweetened solutions. Non-dependent and non- fluid or non-food deprived rats will choose ethanol over water in limited access situations and will lever press to ingest sufficient quantities to produce meaningful blood ethanol levels. Work in the previous funding period was directed at examining the role of a limbic-nucleus accumbens-extrapyramidal circuit in ethanol reinforcement. Results show that while dopamine in the nucleus accumbens may modulate ethanol reinforcement, other systems such as GABA and glutamate may have important if not crucial roles. The purpose of the present proposal will be to extend these earlier results in non-dependent rats to other brain sites such as the amygdala and to other neurotransmitter systems. A further purpose will be to examine the neuropharmacological basis of ethanol reinforcement in dependent rats. Finally, possible sex differences in ethanol reinforcement and the neuropharmacological modulation of ethanol reinforcement will be explored. Non-food or non-fluid deprived male and female rats will be trained to self-administer ethanol in daily 30 minute sessions using a saccharin fade out procedure. The rats then will be implanted with bilateral cannulas aimed at specific brain sites implicated by previous work on ethanol reinforcement. Following establishment of stable ethanol self-administration, the rats will be microinjected with neurotransmitter antagonists and agonists prior to ethanol self-administration sessions. Dose effect functions will be generated using Latin square within subjects designs for dopamine, serotonin, opioid peptide, GABA and glutamate antagonists, and in some cases agonists. Experiments yielding significant results will be repeated in control experiments where rats are trained to self-administer saccharin (without ethanol). Following establishment of critical sites and neuropharmacological effects in non-dependent rats, similar experiments will be repeated in dependent animals and in female rats. The hypothesis under test is that chronic ethanol may result in adaptive changes in those brain mechanisms critical for ethanol reinforcement in the non-dependent animal and that this adaptation may be a crucial motivational part of the dependence process. Knowledge of the neurobiological basis of the motivational aspects of ethanol dependence will provide key information for the development of diagnosis, treatment, and prevention strategies for alcoholism.

This proposal was generated in response to RFA DA-93-01 and is designed to establish in vivo neurochemical and electrophysiological correlates of cocaine-seeking behaviors that can then be used to design potential novel treatment drugs for cocaine dependence. These animal models are designed to permit assessment of the neurochemical and physiological basis of cocaine reward magnitude, strength of cocaine-seeking behaviors ("craving"), and the "propensity to relapse" in limited access and the dependent state. Behavioral parameters of various cocaine-seeking behaviors will be established in rats with different histories of cocaine preexposure. Specifically, cocaine-seeking behaviors of rats with a history of daily limited-access cocaine self-administration will be compared to those exhibited by rats given extended access to cocaine. The proposed studies will examine whether the behavioral manifestations of cocaine "craving" are subserved by the same or different, neuropharmacological substrates as the acute reinforcing effects of cocaine. To accomplish these goals, behavioral parameters of responding for cocaine under different reinforcement contingencies will be established in rats after episodes of extended-access cocaine self- administration using progressive ratio, and multiple schedules. Repeated extinction and spontaneous recovery tests in the presence and absence of environmental cues associated with cocaine availability will provide information about the persistence of cocaine-seeking behaviors. Neurochemical correlates of these behavioral measures will be established using intracranial microdialysis for dopamine and serotonin in various forebrain sites. Neurophysiological correlates of these behavioral methods will be measured using extracellular recording techniques in freely-moving rats at various forebrain sites. Novel pharmacological therapies will be developed by determining effective pharmacological means of reversing the neurochemical (in vivo microdialysis) and electrophysiological (extracellular recording) changes associated with these various components of cocaine-seeking behavior in both limited-access and dependent animals. Treatments effective in modifying these neuropharmacological events will then be tested in the behavioral paradigms for potential pharmacotherapeutic action. These studies will provide important, presently unavailable insights into the neuropharmacological substrate(s) mediating specific cocaine-seeking behaviors as well as information about the relationship between self-administration history and cocaine-seeking behaviors. Finally, the work in this proposal will provide a novel means for the development of specific and novel drugs with therapeutic potential for treating or preventing cocaine abuse and dependence.

Under this NIAAA Center grant The Scripps Research Institute Alcohol Research Center proposes to continue its long-term, cooperative interdisciplinary research program focused on the theme of the central nervous system effects of alcohol. For this renewal application, the TSRI- ARC will consist of 9 components. The Administrative Core resource (supervising six supporting resource units) and 7 independent collaborating research components as well as a separate Pilot Project Component are designed to stimulate new research directions and recruit new investigators to ethanol research. The TSRI-ARC research program remains strongly interdisciplinary, employing molecular, cellular, and cellular system and behavioral research methods on experimental animal subjects and human subjects. Three major subthemes emerging from our past 19 years of work within this Center will be continued: 1) the molecular, cellular, system mechanisms of the acute reinforcing actions of ethanol (intoxication) and its potential molecular, neurophysiologic, neuroendocrine, and behavioral concomitants (including reinforcing and aversive effects of acute intoxication, (Drs. Sanna, Gruol, Siggins, Henriksen, Rivier, and Weiss), 2) the molecular, cellular and system changes underlying the adaptive responses to the brain systems involve in ethanol reinforcement during alcohol dependence following chronic exposure (Drs. Sanna, Gruol, Siggins, Henriksen, Rivier, and Weiss, 3) the molecular, cellular and neuropsychological basis for susceptibility to alcohol dependence and relapse in humans (Drs. Sanna, Gruol, Siggins, Henriksen, Rivier, Weiss, Ehlers and Polich). Progress during the previous funding period including new advances in animal model development have lead to a focusing of the new proposed work to understanding the neurobiology of ethanol dependence and relapse (subthemes 2 and 3). Individual research components range from studies of the effects of chronic ethanol on neuroadaptive mechanisms involving neuronal transduction mechanisms at the cellular, neuroendocrine and neurobehavioral levels to clinical studies of vulnerability to dependence. The TSRI-ARC also supports 11 RO1, R20, R29, and U10 grants of the TSRI- ARC PIs, and other major alcohol research groups at the UCSD VA Medical Center and San Diego State University through the collaboration, training and information exchange of the Administrative Core Support Units. Research funded within this Center provides the basic research knowledge required from development of better understanding of the basis for susceptibility to alcoholism important for new treatments and prevention.

Acutely ethanol, has tension-reducing (anxiolytic) properties, whereas chronic ethanol, administered to the point of dependence, induces anxiety particularly during withdrawal. Both of these actions may contribute to the motivational effects important for the development and maintenance of ethanol abuse. Previous work in our laboratory has established sensitive measures for the anti-conflict and dependence-producing actions of ethanol. Neuropharmacological studies have provided evidence for a role of GABA, serotonin and corticotropin-releasing factor (CRF) in these anti-conflict and dependence-inducing effects of ethanol, and neurobiological sites for the effects of ethanol on anxiety substrates may involve the amygdala. The purpose of the present proposal will be to continue to explore the neurobiological basis for the anti-conflict ("anxiolytic") and dependence-inducing effects of ethanol. Emphasis will be placed on determining the brain sites of interaction of ethanol with GABA, serotonin and CRF. Studies will also be directed at exploring a possible role of neural and adrenal steroids and sex differences in those effects of ethanol. The anti-conflict effects of acute ethanol will be measured in a Geller-Seifter conflict test sensitive to ethanol. Ethanol withdrawal will be measured with three automated tests: The elevated plus maze, as an index of "anxiety"; the acoustic startle response as a measure of "arousal"; and a "tremor response" as a quantifiable measure of physical withdrawal. Confirmation of brain site specific neurochemical interactions with ethanol will be made with in vivo microdialysis in awake, freely moving animals exposed to acute or chronic ethanol. Results derived from these studies will go far towards identifying specific neurochemical brain systems of critical importance for the development and maintenance of ethanol abuse and alcoholism.

DESCRIPTION: (Applicant's Abstract) Animal models exist for many components of the psychostimulant dependence cycle including the acute reinforcing effects of cocaine and amphetamine, withdrawal from these drugs and even reinstatement of responding after extinction. Basic preclinical studies have implicated brain dopamine neurotransmission in all three of these components of the psychostimulant dependence cycle. However, attempts to modify these aspects of cocaine dependence by manipulation of brain dopamine systems with agonists or antagonists have met with limited success at the clinical level prompting the search for novel means of altering dopaminergic function. Dopamine partial agonists have the unique neuropharmacological profile of having high affinity for the receptor, but low intrinsic activity. The functional consequence is that partial agonists may act as agonists in conditions of low receptor occupancy by the transmitter as, for example, in the case of denervation or depletion due to overstimulation of activity. Similarly, the same compounds may act as functional antagonists in case of high endogenous transmitter tone as may happen during intense presynaptic activity or after pharmacological stimulation (e.g. exposure to cocaine or amphetamine). Recent studies in our laboratories have shown that two partial agonists SDZ 208-911 and terguride can increase cocaine self-administration (decrease the inter-injection interval) in rat, similar to the effects observed with dopamine antagonists. Unknown. however, is the nature of the interaction of these partial agonists with the full cocaine dose-effect function and with other aspects of psychomotor stimulant dependence. The purpose of the proposed studies is to explore the effects of a wide range of dopamine partial agonists on intravenous cocaine self-administration, cocaine withdrawal and the reinstatement of cocaine self-administration after extinction. Four doses of different partial agonists will be tested in rats against full dose effect functions for cocaine and amphetamine self-administration and on a multiple schedule for food and drug (Specific Aims 1&2). The effect of dopamine partial agonists will be tested on cocaine withdrawal using locomotor activity and brain stimulation reward thresholds (Specific Aim 3) and on the reinstatement of cocaine self-administration in animals subjected to extinction of cocaine self-administration (Specific Aim 4). These studies will help elucidate the effectiveness of dopamine partial agonists with a wide range of intrinsic efficacies in modifying various phases of the natural history of psychostimulant dependence.

DESCRIPTION: The purpose of this program is to train promising young scientists at the postdoctoral level in the mulidisciplinary strategies of molecular, cellular and behavioral neuropsychopharmacology. Present day research on alcohol and other abused drugs requires methodologies that can pinpoint molecular and cellular sites in the brain, and integrate these mechanisms at the functional system level of analysis. The Scripps Alcohol Research Center (ARC), combined with a very active independent but interactive San Diego Alcohol Research Program, has developed methods for combining biochemical, morphological, physiological and behavioral research to arrive at broad based studies of sites and mechanisms of drug action. These methods permit the pursuit of research lines broadly through such fields as molecular biology, light and electron microscopic histology, immununo- cytochemistry, electrophysiologic analysis (in vivo and in vitro), tissue culture, neuroendocrinology, behavioral pharmacology, the correlation of animal behavior and neuronal discharge patterns, electroencephalography and event related cortical evoked potential recordings applied to animal and human subjects. In addition to developing those technical skills in our trainees deemed essential for their transition into career scientists, we also attempt to develop other important skills: creative research expression, critical selection of problems, data recording, validation and security, data interpretation and manuscript and grant preparation. Through our resident senior program staff, visiting senior scientists and courses given in conjunction with local educators, trainees are exposed to a variety of research applications and techniques. Trainees participate in frequent scientific project review conferences, research seminars and journal clubs as well as actively participating in research programs and manuscript review. Each trainee will also receive a course on ethanol neuropsychopharmacology and a course in conduct of research. Postdoctoral fellows and other advanced trainees are selected with preferential weighting towards those seeking interdisciplinary methodologies. They are then assigned to one of the Principal Investigators responsible for Individual Research Components of the Scripps ARC and the San Diego Alcohol Research community, who will coordinate their initial research project selections. Depending upon a trainee's prior research skills, collaborations with more than one senior scientist are encouraged.

This proposal continues to be directed at identifying and characterizing the behavioral, neurochemical, and neuropharmacological mechanisms involved in ethanol-seeking behavior and dependence. Studies conducted during the previous funding period have characterized neuropharmacological mechanisms involved in the anti-punishment effects of ethanol and the motivational effects of ethanol withdrawal. The present proposal extends these efforts by shifting emphasis to the investigation of relapse. Specifically, the objective of this proposal is to employ the alcohol deprivation effect (ADE) as a model of excessive drinking or "loss of control" and to study its neurochemical and neuropharmacological basis. The hypothesis is that alcohol deprivation after a period of chronic ethanol self-administration leads to changes in the functional activity and/or sensitivity to alcohol of the same brain neurotransmitter systems that have previously been implicated in the reinforcing effects of ethanol, the genetic basis of alcohol preference, or the motivational effects of ethanol withdrawal. This hypothesis will be tested by focusing on both pre synaptic and post synaptic changes associated with the ADE using in vivo neurochemical as well as behavioral pharmacological approaches. Specific Aim 1 will examine changes in basal neurotransmitter profiles after alcohol deprivation as well as changes in neurotransmitter sensitivity to systemically injected and self-administered ethanol. Target brain sites will include the shell of the nucleus accumbens and the central nucleus of the amygdala. The neurochemical systems of interest are gamma-aminobutyric acid (GABA), dopamine (DA), endogenous opioids, and corticotropin-releasing factor (CRF). These studies will be complemented by investigations of the behavioral effects of pharmacological agents on deprivation-enhanced ethanol intake in Specific Aim 2. Specific Aim 3 will study the neurobiological basis of the enhancement of the ADE associated with protracted abstinence in previously ethanol-dependent rats. These experiments are designed to determine whether the ADE in post-dependent rats involves identical, but more pronounced changes in neurotransmitter function compared to non-dependent rats, or involves additional or different neurochemical mechanisms. Finally, Specific Aim 4 will begin to explore sex differences and their neurobiological basis in the susceptibility to the ADE. The results of these studies are expected to reveal adaptive processes that lead to enhanced alcohol intake after deprivation and, thereby, may provide novel insights into brain mechanisms that underlie processes such as alcohol craving, loss of control, and relapse.

DESCRIPTION (Adapted From The Applicant's Abstract): This is a competing renewal application to continue to study the neural mechanisms of opiate reinforcement and dependence. Work during the previous funding period has established animal models of heroin self-administration in dependent animals and animal models of conditioned withdrawal in dependent animals. Studies have begun to elucidate the neurobiological substrates for these effects with a focus on a basal forebrain continuum from the shell of the nucleus accumbens to the central nucleus of the amygdala termed the "extended amygdala." Sites in the shell of the nucleus accumbens, bed nucleus of the stria terminalis and central nucleus of the amygdala appear particularly important for processing the aversive stimulus effects of opiate dependence independent of somatic effects. Preliminary neuropharmacological studies also suggest a potential role for corticotropin-releasing factor in mediating such aversive stimulus effects. The purpose of the present proposal is to test the hypothesis that specific neuropharmacological elements in the extended amygdala contribute to the motivational effects associated with the development and maintenance of opiate dependence. To test this hypothesis, systematic neuropharmacological studies will be employed to examine the role of CRF, serotonin, glutamate, and dopamine in the reinforcing effects of heroin using intravenous self-administration in dependent rats (Specific Aim 1). In addition, the neuropharmacological basis for the aversive stimulus effects of opiate withdrawal in dependent rats will be explored using place conditioning (place aversion-- Specific Aim 2). An attempt will be made to determine the neural substrates for the expression of opiate withdrawal-induced place aversion by cell body-selective neurotoxin lesions of the regions projecting to the extended amygdala and with lesions of the extended amygdala itself (Specific Aim 3). Finally, the neuropharmacological basis within the extended amygdala itself for heroin self-administration in dependent rats and opiate withdrawal-induced place aversion will be explored using microinjections of neurotransmitter agonists and antagonists directly into sites within the extended amygdala continuum (Specific Aim 4). These studies will go far towards elucidating the neural mechanisms mediating the brain changes critical for the development of opiate dependence and as such will provide basic information important for establishing vulnerability to opiate addiction and for establishing novel prevention and treatment interventions.

DESCRIPTION: (Adapted from the Investigator's Abstract) This proposal seeks to extend the focus of investigation conducted during the previous funding period, which was concerned with the effects of dopamine D2 partial agonists on multiple measures of cocaine self-administration and withdrawal, to study the neuropharmacology of methamphetamine dependence. Work during the previous funding period has allowed the characterization of the effects of dopamine D2 partial agonists on both cocaine and amphetamine self-administration under conditions of limited daily access and across full dose effect functions. In addition, the addictive liability of partial agonists has been examined showing that terguride, a prototype dopamine D2 partial agonist, does not function as a substrate for self-administration and does not act as a priming agent reinstating extinguished self-administration. Furthermore, initial studies have addressed the hypothesis that partial dopamine agonists may also act as candidates for pharmacotherapy for amphetamine and methamphetamine dependence. The aim of the proposed studies is to further characterize the effects of partial dopamine agonists acting on D1, D2 and D3 dopamine receptor subtypes on methamphetamine dependence. For this purpose, a variety of animal tests tailored to model several components of the methamphetamine dependence cycle will be employed. The guiding hypothesis of the present proposal is that dopamine partial agonists may provide innovative pharmacological measures of interaction with multiple aspects of methamphetamine-seeking behavior that may ultimately lead to the loss of control which represents the cardinal feature of the psychostimulant addictive cycle. A detailed characterization of the effects of dopamine partial agonists on multiple measures of methamphetamine self-administration will allow insight into the effects of these drugs on the acute reinforcing properties of methamphetamine in Specific Aim 1. Specific Aim 2 and 3 will examine the abstinence phase investigating the potential of partial agonists to prevent different behavioral changes associated with withdrawal. The potential of partial agonists to induce relapse or prevent methamphetamine-induced relapse of methamphetamine-seeking behavior will also be evaluated. Specific Aim 4 will then characterize the transition from moderate to excessive methamphetamine-seeking behavior and investigate whether partial dopamine agonists modify drug intake in rats with a history of drug escalation.

DESCRIPTION (provided by applicant): This is a competing renewal application for a Consortium for the Integrative Neuroscience Initiative on Alcoholism (INIA)-West (Notice# RFA-AA-11-006) to identify the molecular, cellular, and behavioral neuroadaptations that occur in specific brain neurocircuitries that result in excessive alcohol consumption. This multidisciplinary initiative focuses on the molecular and cellular neuroadaptations in brain addiction circuits associated with the basal forebrain, including reward circuitry in the ventral striatum and dependence circuitry in the extended amygdala. The overall hypothesis for INIA-West is that genetic differences and neuroadaptations in reward circuitry are responsible for individual differences in the vulnerability to the excessive alcohol consumption. Sixteen research components and six scientific cores will use excessive drinking models in animals to mimic the binge- and dependence-induced excessive drinking of alcohol abuse disorders. The overall goals of INIA-West are (1) to confirm gene targets nominated by expression assays or other methods by use of transgenic, knockout, inducible knockout, site-specific knockout, RNAi, in situ hybridization, in vivo electrophysiology, in vivo imaging, and next-generation sequencing, (2) to identify druggable targets that are most promising for medication development for the treatment of alcoholism by use of novel molecules in concert with molecules with existing FDA approval in animal models with the most predictive ability, and (3) to attract new and innovative investigators to the field of alcohol research by recruiting individuals for U01 grants and Pilot projects and by making the informatics integrated datasets accessible, searchable, and interactive with other databases for all scientists interested in alcoholism research. Core facilities are proposed that provide molecular genetic support and target assessment translation for medications target development. A Pilot Project program is proposed to identify exciting new areas of research and the continual recruitment of new investigators to the alcohol field. The INIA program will be directed by the Administrative Core in close cooperation with the Executive Committee and Steering Committee and with the continual advice of a distinguished Scientific Advisory Board.

DESCRIPTION (provided by applicant): The purpose of this program is to train promising young scientists at the postdoctoral level in the multi disciplinary strategies of molecular, cellular, behavioral, and clinical neuropsychopharmacology. Research on alcohol and other abused drugs requires methodologies that can pinpoint molecular and cellular sites in the brain, and integrate these mechanisms at the system level of analysis. The Scripps Alcohol Research Center (ARC), combined with a very active independent but interactive San Diego Alcohol Research Community, has developed methods for combining biochemical, morphological, physiological and behavioral research to arrive at broad based studies of sites and mechanisms of drug action. These methods permit the pursuit of research lines broadly through such fields as molecular biology, immunocytochemistry, electrophysiologic analysis (in vitro and in vivo), neuroendocrinology, behavioral pharmacology, electroencephalography and event related cortical evoked potential recordings applied to animal and human subjects, and clinical studies. In addition to developing those technical skills in our trainees deemed essential for their transition into career scientists, we also attempt to develop other important skills: creative research expression, critical selection of problems, experimental design, data recording, validation and security, data interpretation, manuscript and grant preparation, and ethical conduct of research. Through our resident senior program staff, visiting senior scientists and courses given in conjunction with local educators, trainees are exposed to a variety of research applications and techniques. Trainees participate in scientific project review conferences, research seminars and journal clubs is well as actively participating in research programs. Each trainee will also receive a course on ethanol neuropsychopharmacology and a course in ethical conduct of research. Both informal and formal recruitment mechanisms are well established and have provided a sufficient number of high quality applicants. Extensive programs are in place for recruitment of minority and for facilitating minority interest in alcohol research. Postdoctoral fellows and other advanced trainees are selected with preferential weighting towards those seeking interdisciplinary methodologies. They are then assigned to one of the Principal Investigators of the training grant who will coordinate their initial research project selections. Depending upon a trainee's prior research skills, collaborations with more than one senior scientist are encouraged.

DESCRIPTION (provided by applicant): Animal models are being used throughout The Scripps Research Institute-Alcohol Research Center (TSRI-ARC) and Center at Large to pursue then neurobiological basis of ethanol intoxication, dependence, and vulnerability to relapse. Work in the Animal Model Development Unit of the Alcohol Research Center grant during the previous funding period has provided the foundation for the animal models that are being used throughout the Center. In the present proposal, the former Animal Model Development Unit has been assimilated into a Core component of the TSRI-ARC and the Center at Large. This is necessitated by the major advances in development of animal models during the previous funding period and the reliance of the animal components of the Center and the Center at Large on this facility. The Animal Model Core will serve 3 purposes for the proposed TSRI-ARC and Center at Large: 1) The Animal Model Core will provide the facilities for inducing dependence using the ethanol vapor chambers for all investigators in the TSRI-ARC and Center at Large, 2)The Animal Model Core will provide blood ethanol levels and vaginal smear data for TSRI-ARC and Center at Large, 3) The Animal Model Core will continue to refine the animal models of dependence to be utilized by the Center including exploring different lengths and amounts of ethanol exposure as well as the influence of gender and genetic variables. The Specific Aims of the Animal model development program are to: 1). Examine intermittent versus continuous ethanol vapor exposure models as a means to optimize allostatic processes, 2). Characterize ethanol self-administration in dependent female rats with respect to estrous cycle phase, 3). Examine genetic susceptibility to ethanol self-administration in dependent rats selectively bred for differential ethanol drinking, and 4). Examine genetic susceptibility to ethanol self-administration in dependent rats selectively bred for traits postulated to involve allostatic mechanisms. The service and animal model development proposed for this Core are a critical integrative part of the TSRI-ARC and Center at Large that provide a major contribution to the "centerness" of the program by standardization of animal models, collaborative interactions and communication among investigators.

The overall objective of the Research Translation/Information Dissemination (Education) Component of The Scripps Research Institute Alcohol Research Center (TSRI ARC) is to advance the NIAAA¿s mission to translate and disseminate scientific research findings to researchers, healthcare professionals, policy makers, and the general public. The main goals of the Research Translation/Information Dissemination Component are to: 1) develop and evaluate an education program to the high risk Mexican American minority and EuroAmerican communities in San Diego greatly in need of tools to combat underage drinking and alcohol use disorders in their communities, and 2) expand the dissemination of knowledge about the scientific findings of the TSRI ARC and their translation to clinical alcohol diagnosis, treatment, and prevention to students, scientific and medical professionals and trainees, and the lay public. To accomplish these goals, we will perform research translation/Information Dissemination (Education and Outreach) to High Risk Mexican American Immigrant and Other Communities in San Diego (Specific Aim 1), perform Integration, Translation, Training, and Education in Alcohol Research to ARC scientists and trainees, undergraduates, psychology, pharmacy graduate students, and addiction professionals (Specific Aim 2), and effect the TSRI ARC High School/College/Graduate School Intern Program (Specific Aim 3). Finally, we will disseminate the scientific findings of the TSRI ARC, information on alcohol use disorders, and how recent neurobiological research is informing new approaches to clinical alcoholism to the general public (Specific Aim 4).

DESCRIPTION (Provided by applicant): The Alcohol Research Center of The Scripps Research Institute (TSRI-ARC) proposes to continue its interdisciplinary program focused on the theme of the central nervous system effects of alcohol. For this renewal application, the TSRI-ARC will be a P60 consisting of 9 components plus an Educational Component. Four core components are proposed: Administrative, Animal Models Development, Biochemical and Pilot. Five research components are proposed: Cellular Neurobiology (Roberto/Siggins), Neuroendocrinology (Rivier), Neuropharmacology Neuropeptides (Zorrilla/Weiss), Neuropharmacology Endocannabinoids (Parsons), and Clinical Neurobehavioral (Ehlers). The overall hypothesis of the TSRIARC is that the function of specific neurotransmitter synapses in select parts of the reward and stress systems that are compromised by chronic ethanol administration account for the development of vulnerability to alcoholism in genetically prone individuals. Two major themes have emerged from our present research 1) the neuropharmacological mechanisms of vulnerability to dependence depend on how specific brain reward and stress circuits change during the transition from initiation of drinking to binge drinking to dependence (Specific Aims 1 &2);2) the neuropharmacological changes produced by binge drinking in adolescents and young adults may drive excessive drinking and vulnerability to dependence in adults (Specific Aim 3). Progress during the previous funding period has led to a focus on understanding dependence-induced neuroadaptive mechanisms and residual allostatic changes that persist following acute abstinence. The new focus of the ARC is on neuroadaptive changes that are engaged by binge drinking that form a transition from initiation of drinking to dependence and form a crucial part of the basis for inheritable susceptibility to human alcoholism. The TSRI-ARC also supports the Center at Large, which includes: seventeen NIAAA R01's, two U01's, four NIAAA R21's, two NIAAA R37 awards, one NIAAA T32 training grant, one NIAAA K01 award and one NIAAA K99 award. Training and information dissemination to the San Diego community will be effected by the training opportunities of the Center including an NIAAA training grant and the Education Component.

DESCRIPTION (provided by applicant): This is a competing renewal application to continue to study the neural mechanisms of opiate reinforcement and dependence. Work during the previous funding period has established animal models of the motivational effects of opiate withdrawal (place aversion), animal models of heroin self-administration in dependent rats and animal models of conditioned increases in responding to precipitated opiate withdrawal. Using these models, studies have identified critical elements in the brain stress systems in the basal forebrain (corticotropin releasing factor, neuropeptide Y and norepinephrine) in the motivational effects of opiate withdrawal. The purpose of the present proposal is to test the hypothesis that the brain stress neurotransmitter systems: corticotropin-releasing factor, norepinephrine and neuropeptide Y may have critical roles in mediating the aversive stimulus effects of opiate withdrawal, the development of motivational aspects of opiate dependence, and the motivational effects of conditioning associated with the aversive stimulus effects of opiate withdrawal. A sub-hypothesis is that previously neutral stimuli that acquire motivational significance through pairing with opiate withdrawal utilize specific neural circuits associated with input to the extended amygdala. To test these hypotheses, the animal models of intravenous self-administration in dependent animals will be validated (Specific Aim 1), the neuropharmacological mechanisms within specific sites of the extended amygdala involved in the aversive motivational state of opiate withdrawal (place aversion) will be explored (Specific Aim 2), the neuropharmacological mechanisms within specific sites within the extended amygdala involved in heroin self-administration in dependent rats will be explored (Specific Aim 3) and the neuroanatomical basis for the conditioned increases in heroin self-administration produced by precipitated opiate withdrawal will be explored (Specific Aim 4). The present proposal will go towards elucidating the neural circuits and neuropharmacological systems within the basal forebrain circuits which are critical for the motivational effects of opiate dependence and will provide critical information important for identifying the basis for individual differences in vulnerability to opiate addiction and help establish the basis for novel approaches to prevention and treatment.

DESCRIPTION (provided by applicant): This is a competing renewal application to continue to study the neural mechanisms of opiate reinforcement and dependence. Work during the previous funding period has established animal models of the motivational effects of opiate withdrawal (place aversion), animal models of heroin self-administration in dependent rats and animal models of conditioned increases in responding to precipitated opiate withdrawal. Using these models, studies have identified critical elements in the brain stress systems in the basal forebrain (corticotropin releasing factor, neuropeptide Y and norepinephrine) in the motivational effects of opiate withdrawal. The purpose of the present proposal is to test the hypothesis that the brain stress neurotransmitter systems: corticotropin-releasing factor, norepinephrine and neuropeptide Y may have critical roles in mediating the aversive stimulus effects of opiate withdrawal, the development of motivational aspects of opiate dependence, and the motivational effects of conditioning associated with the aversive stimulus effects of opiate withdrawal. A sub-hypothesis is that previously neutral stimuli that acquire motivational significance through pairing with opiate withdrawal utilize specific neural circuits associated with input to the extended amygdala. To test these hypotheses, the animal models of intravenous self-administration in dependent animals will be validated (Specific Aim 1), the neuropharmacological mechanisms within specific sites of the extended amygdala involved in the aversive motivational state of opiate withdrawal (place aversion) will be explored (Specific Aim 2), the neuropharmacological mechanisms within specific sites within the extended amygdala involved in heroin self-administration in dependent rats will be explored (Specific Aim 3) and the neuroanatomical basis for the conditioned increases in heroin self-administration produced by precipitated opiate withdrawal will be explored (Specific Aim 4). The present proposal will go towards elucidating the neural circuits and neuropharmacological systems within the basal forebrain circuits which are critical for the motivational effects of opiate dependence and will provide critical information important for identifying the basis for individual differences in vulnerability to opiate addiction and help establish the basis for novel approaches to prevention and treatment.

[unreadable] DESCRIPTION (provided by applicant): The purpose of this program is to train promising young postdoctoral scientists in the multidisciplinary strategies of molecular, cellular, behavioral, and clinical neuropsychopharmacology. The Scripps Research Institute Alcohol Research Center (ARC), combined with a very active San Diego Alcohol Research Community, has developed a conceptual framework to study the neurobiology of alcoholism and the neurobiological bases for individual differences in vulnerability to alcoholism. Biochemical, morphological, physiological, and behavioral methods have been developed to study the neuropsychopharmacology of alcoholism through such fields as molecular biology, immunocytochemistry, electrophysiology (both in vivo and in vitro), neuroendocrinology, behavioral pharmacology, electroencephalography, cognitive and motivational testing applied to animal and human subjects, and clinical studies. We develop in the trainees important skills for pathways to independence, such as creative research expression, critical selection of problems, experimental design, data recording, validation and security, data interpretation, manuscript and grant preparation, and the ethical conduct of research. Trainees participate in scientific project development and implementation, research seminars, and journal clubs. Each trainee receives formal courses on Ethanol Neuropsychopharmacology and the ethical conduct of research. Both informal and formal recruitment mechanisms are well established and provide a sufficient number of high quality applicants. Extensive programs are in place for recruitment of minority candidates and for facilitating minority interest in alcohol research. Significant success in the past funding period has resulted in the recruitment of five young faculty, including two minority faculty, to the training program. A formal evaluation of the success of the program charts the career development of pathways to independence in alcohol research of former fellows. Postdoctoral fellows and other advanced trainees are selected with preferential weighting given to those seeking interdisciplinary methodologies. Trainees are assigned to one of the training grant faculty members who coordinate their initial research project selections. Depending upon a trainee's prior research skills, collaborations with more than one senior faculty member are encouraged. In summary, the training program discussed herein provides a dynamic environment for fellows to develop a strong foundation to pursue a career in the neuropsychopharmacology of alcoholism. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): This is a competing renewal application to continue to study the neural mechanisms of opiate reinforcement and dependence. Work during the previous funding period has established animal models of the motivational effects of opiate withdrawal (place aversion), animal models of heroin self-administration in dependent rats and animal models of conditioned increases in responding to precipitated opiate withdrawal. Using these models, studies have identified critical elements in the brain stress systems in the basal forebrain (corticotropin releasing factor, neuropeptide Y and norepinephrine) in the motivational effects of opiate withdrawal. The purpose of the present proposal is to test the hypothesis that the brain stress neurotransmitter systems: corticotropin-releasing factor, norepinephrine and neuropeptide Y may have critical roles in mediating the aversive stimulus effects of opiate withdrawal, the development of motivational aspects of opiate dependence, and the motivational effects of conditioning associated with the aversive stimulus effects of opiate withdrawal. A sub-hypothesis is that previously neutral stimuli that acquire motivational significance through pairing with opiate withdrawal utilize specific neural circuits associated with input to the extended amygdala. To test these hypotheses, the animal models of intravenous self-administration in dependent animals will be validated (Specific Aim 1), the neuropharmacological mechanisms within specific sites of the extended amygdala involved in the aversive motivational state of opiate withdrawal (place aversion) will be explored (Specific Aim 2), the neuropharmacological mechanisms within specific sites within the extended amygdala involved in heroin self-administration in dependent rats will be explored (Specific Aim 3) and the neuroanatomical basis for the conditioned increases in heroin self-administration produced by precipitated opiate withdrawal will be explored (Specific Aim 4). The present proposal will go towards elucidating the neural circuits and neuropharmacological systems within the basal forebrain circuits which are critical for the motivational effects of opiate dependence and will provide critical information important for identifying the basis for individual differences in vulnerability to opiate addiction and help establish the basis for novel approaches to prevention and treatment.

This is a supplemental proposal to the Consortium for the Integrative Neuroscience Initiative on Alcoholism[unreadable] West (INIA) (original RFA: AA-01-002) to provide funds for an innovative project important the goals of INIA west. The overall goals of the Consortium are to identify the molecular, cellular, and behavioral neuroadaptations that occur in specific brain neurocircuitries that result in excessive alcohol consumption. More specifically, the focus of this multidisciplinary initiative is on the molecular and cellular neuroadaptations in the brain reward circuits responsible for the individual differences in vulnerability to the excessive consumption of alcohol. The proposed studies of the supplement provide highly innovative molecular biological approaches to two key INIA-related questions, namely where ethanol acts to produce dependence and which genes are implicated in this process. To accomplish these goals specific ethanol- activated brain sites using innovative reporter mouse models visualizing neuronal activation in vivo will be identified (Specific Aim 1) and novel genes regulated by alcohol in the extended amygdala will be identified (Specific Aim 2) using highly innovative, novel molecular biological approaches and established INIA west models of excessive ethanol intake. In addition, an NIH/INSERM International Associated laboratory between the Committee on the Neurobiology of Addictive Disorders chaired by George Koob (The Scripps Research Institute, La Jolla, California, USA) and the Department of Neurobiology directed by Brigitte Kieffer (IGBCM, Strasbourg, France) will be established. The Kieffer and Koob laboratories have engaged in a highly successful collaboration for more than 10 years and have published together extensively lending support for the proposed extension of their ongoing collaboration. The proposed studies will further the goals of the INIA west consortium by providing not only exciting and unique novel scientific approaches but also by recruiting new talented investigators to the study of neurobiology of alcohol abuse disorders.

12-20 years old; 21+ years old; Absolute ethanol; Abstinence; Acute; Adolescence; Adolescent; Adolescent Youth; Adult; Age Factors; Alcohol Drinking; Alcohol Intoxication; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohol, Ethyl; Alcoholic Intoxication; Alcoholism; Alcohols; Animal Model; Animal Models and Related Studies; Animals; Blood; Blood Alcohol Content; Blood alcohol level measurement; Characteristics; Chemical Class, Alcohol; Chronic; Common Rat Strains; Communities; Consumption; Data; Dependence; Development; Drunkenness; Drunkennesses; ETOH; Estrous Cycle; EtOH drinking; EtOH intoxication; Ethanol; Ethanol dependence; Female; Foundations; Funding; Gender; Genetic; Genetic Predisposition; Genetic Predisposition to Disease; Genetic Susceptibility; Goals; Grain Alcohol; Grant; Guidelines; Heavy Drinking; History; Human, Adult; In element; Indium; Inherited Predisposition; Inherited Susceptibility; Investigators; Mammals, Rats; Methylcarbinol; Modeling; Neurobiology; Pap Test; Pap smear; Papanicolaou Smear; Papanicolaou Test; Phase; Process; Purpose; Rat; Rattus; Recording of previous events; Relapse; Relative; Relative (related person); Reliance; Research; Research Institute; Research Personnel; Researchers; Reticuloendothelial System, Blood; Self Administration; Self-Administered; Smears, Cervical; Solutions; Staging; Testing; Time; Vaginal Smears; Withdrawal; Work; addiction; adolescence (12-20); adult human (21+); alcohol addiction; alcohol dependency; alcohol effect; alcohol ingestion; alcohol intake; alcohol product use; alcohol research; alcohol use; alcohol withdrawal; alcohol-dependent; alcoholic beverage consumption; alcoholic drink intake; allostases; allostasis; base; binge alcohol consumption; binge drinking; blood alcohol concentration; blood alcohol level; cervical/vaginal smear; drink heavily; drinking; episodic drinking; ethanol addiction; ethanol consumption; ethanol dependency; ethanol drinking; ethanol effect; ethanol ingestion; ethanol intake; ethanol intoxication; ethanol product use; ethanol research; ethanol use; ethanol withdrawal; ethanol-dependent; etoh use; excess alcohol consumption; excess alcohol ingestion; excess ethanol ingestion; excessive alcohol consumption; excessive alcohol ingestion; excessive alcohol intake; excessive drinking; excessive ethanol ingestion; extreme drinking; genetic etiology; genetic mechanism of disease; genetic vulnerability; heavy alcohol use; juvenile; juvenile human; model organism; neurobiological; teenage; vapor; withdrawal from alcohol

2-arachidonoyl-glycerol; 2-arachidonoylglycerol; 2-arachidonyl-glycerol; 2-arachidonylglycerol; 4-Aminobutanoic Acid; 4-Aminobutyric Acid; 5,8,11,14-Eicosatetraenamide, N-(2-hydroxyethyl)-, (all-Z)-; 5,8,11,14-Eicosatetraenoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester, (5Z,8Z,11Z,14Z)-; 5,8,11,14-eicosatetraenamide, N-(2-hydroxyethyl)-; 5,8,11,14-eicosatetraenoylethanolamide; Absolute ethanol; Acute; Affective; Alcohol Drinking; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohol, Ethyl; Alcoholic; Aminalon; Aminalone; Amygdala; Amygdaloid Body; Amygdaloid Nucleus; Amygdaloid structure; Animals; Anti-Anxiety Agents; Anti-Anxiety Drugs; Anxiety; Anxiolytic Agents; Anxiolytics; Attenuated; Back; Behavior; Boozer; Brain; Brain region; Butanoic acid, 4-amino-; Cannabinoids; Cannabinoids, Endogenous; Cell Communication and Signaling; Cell Nucleus; Cell Signaling; Cellular Neurobiology; Chronic; Common Rat Strains; Corticosterone; Data; Dependence; Dependent drinker; Depression; Disruption; Dorsum; Drugs; ETOH; Emotional; Encephalon; Encephalons; Endocannabinoids; EtOH drinking; Ethanol; Ethanol dependence; Event; Funding; GABA; Genetic; Glutamates; Grain Alcohol; Heavy Drinking; Human; Human, General; Individual; Intracellular Communication and Signaling; L-Glutamate; Lateral; Light; Literature; Maintenance; Maintenances; Mammals, Mice; Mammals, Rats; Man (Taxonomy); Man, Modern; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Medication; Melia; Mental Depression; Methylcarbinol; Mice; Microdialysis; Monitor; Murine; Mus; N arachidonoyl 2 hydroxyethylamide; N-(2-hydroxyethyl)arachidonamide; Negative Reinforcements; Nerve Cells; Nerve Impulse Transmission; Nerve Transmission; Nerve Unit; Nervous System, Brain; Neural Cell; Neurobiology, Cellular; Neurochemistry; Neurocyte; Neuronal Transmission; Neurons; Nucleus; Nucleus Accumbens; Other Finding; Pharmaceutic Preparations; Pharmaceutical Preparations; Phenotype; Photoradiation; Physiologic; Physiological; Play; Prefrontal Cortex; Pregn-4-ene-3,20-dione, 11,21-dihydroxy-, (11beta)-; Rat; Rate; Rats, Wistar; Rattus; Receptor Protein; Regulation; Reinforcements, Negative; Relapse; Research; Role; Science of neurochemistry; Self Administration; Signal Transduction; Signal Transduction Systems; Signaling; Stress; Symptoms; System; System, LOINC Axis 4; Testing; Thinking; Thinking, function; Tranquilizing Agents, Minor; Transmission; Wistar Rats; Withdrawal; Work; alcohol addiction; alcohol dependency; alcohol exposed; alcohol exposure; alcohol ingestion; alcohol intake; alcohol product use; alcohol use; alcohol withdrawal; alcohol-dependent; alcoholic beverage consumption; alcoholic drink intake; amygdaloid nuclear complex; anandamide; anandamide (20.4,n-6); antianxiety agent; arachidonoyl ethanolamide; arachidonoylethanolamide; arachidonylethanolamide; base; behavior test; behavioral pharmacology; behavioral test; biological signal transduction; brain tissue; drink heavily; drinking; driving force; drug/agent; ethanol addiction; ethanol consumption; ethanol dependency; ethanol drinking; ethanol exposed; ethanol exposure; ethanol ingestion; ethanol intake; ethanol product use; ethanol use; ethanol withdrawal; ethanol-dependent; etoh use; excess alcohol consumption; excess alcohol ingestion; excess ethanol ingestion; excessive alcohol consumption; excessive alcohol ingestion; excessive alcohol intake; excessive behavior; excessive drinking; excessive ethanol ingestion; experiment; experimental research; experimental study; exposed to alcohol; exposure to alcohol; extracellular; extreme drinking; gamma-Aminobutyric Acid; heavy alcohol use; in vivo; insight; interstitial; neural mechanism; neurochemistry; neuromechanism; neuronal; neurotransmission; problem drinker; receptor; research study; response; restraint; restraint stress; social role; transmission process; withdrawal from alcohol

2-arachidonoyl-glycerol; 2-arachidonoylglycerol; 2-arachidonyl-glycerol; 2-arachidonylglycerol; 4-Aminobutanoic Acid; 4-Aminobutyric Acid; 5,8,11,14-Eicosatetraenamide, N-(2-hydroxyethyl)-, (all-Z)-; 5,8,11,14-Eicosatetraenoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester, (5Z,8Z,11Z,14Z)-; 5,8,11,14-eicosatetraenamide, N-(2-hydroxyethyl)-; 5,8,11,14-eicosatetraenoylethanolamide; Absolute ethanol; Abstinence; Acute; Agonist; Alcohol Drinking; Alcohol Intoxication; Alcohol consumption; Alcohol dependence; Alcohol, Ethyl; Alcoholic Intoxication; Alcohols; Aminalon; Aminalone; Amygdala; Amygdaloid Body; Amygdaloid Nucleus; Amygdaloid structure; Animal Model; Animal Models and Related Studies; Area; Bed Nucleus of Stria Terminalis; Behavior; Behavioral; Biochemical; Brain; Brain region; Butanoic acid, 4-amino-; Cannabinoids; Cannabinoids, Endogenous; Cell Nucleus; Cells; Chemical Class, Alcohol; Chromosome Pairing; Chronic; Collaborations; Common Rat Strains; Communication; Data; Dependence; Dialysis; Dialysis procedure; Drugs; Drunkenness; Drunkennesses; Dynorphins; ETOH; Electrodes; Encephalon; Encephalons; Endocannabinoids; Enkephalins; EtOH drinking; EtOH intoxication; Ethanol; Ethanol dependence; Exposure to; Frequencies (time pattern); Frequency; Funding; GABA; Glutamates; Grain Alcohol; Heavy Drinking; Hour; Hypothalamic structure; Hypothalamus; In Vitro; Ion Channel; Ionic Channels; L-Glutamate; Ligands; Mammals, Mice; Mammals, Rats; Measures; Mediating; Medication; Membrane; Membrane Channels; Methods; Methylcarbinol; Mice; Microdialysis; Modeling; Molecular; Murine; Mus; N arachidonoyl 2 hydroxyethylamide; N-(2-hydroxyethyl)arachidonamide; Nerve Cells; Nerve Transmitter Substances; Nerve Unit; Nervous System, Brain; Neural Cell; Neurochemistry; Neurocyte; Neurons; Neuropeptide Tyrosine; Neuroregulator; Neurotransmitters; Nucleus; Nucleus Accumbens; Opioid; Opioid Receptor; Pharmaceutic Preparations; Pharmaceutical Preparations; Physical Dialysis; Physiologic pulse; Play; Potentials, Synaptic; Property; Property, LOINC Axis 2; Protocol; Protocols documentation; Psychological reinforcement; Pulse; Pulse taking; Rat; Rattus; Receptor Activation; Receptor Protein; Receptors, Opiate; Reinforcement; Reinforcement (Psychology); Research; Role; Science of neurochemistry; Self-Administered; Site; Slice; Standards; Standards of Weights and Measures; Stria Terminalis Nucleus; Structure of terminal stria nuclei of preoptic region; Synapses; Synapsis; Synapsis, Chromosomal; Synaptic; Synaptic Potentials; System; System, LOINC Axis 4; Testing; Thinking; Thinking, function; Transmission; Week; Withdrawal; alcohol addiction; alcohol dependency; alcohol effect; alcohol exposed; alcohol exposure; alcohol ingestion; alcohol intake; alcohol product use; alcohol reinforcement; alcohol reward; alcohol use; alcohol-dependent; alcoholic beverage consumption; alcoholic drink intake; amygdaloid nuclear complex; anandamide; anandamide (20.4,n-6); arachidonoyl ethanolamide; arachidonoylethanolamide; arachidonylethanolamide; base; binge alcohol consumption; binge drinking; biological adaptation to stress; conditioning; dialysis therapy; drink heavily; drug/agent; episodic drinking; ethanol addiction; ethanol consumption; ethanol dependency; ethanol drinking; ethanol effect; ethanol exposed; ethanol exposure; ethanol ingestion; ethanol intake; ethanol intoxication; ethanol product use; ethanol reinforcement; ethanol reward; ethanol use; ethanol-dependent; etoh use; excess alcohol consumption; excess alcohol ingestion; excess ethanol ingestion; excessive alcohol consumption; excessive alcohol ingestion; excessive alcohol intake; excessive drinking; excessive ethanol ingestion; exposed to alcohol; exposure to alcohol; extreme drinking; gamma-Aminobutyric Acid; heavy alcohol use; hypothalamic; in vivo; membrane structure; model organism; neuroadaptation; neurochemistry; neuronal; neuropeptide Y; nociceptin; orphanin FQ; paired stimuli; patch clamp; postsynaptic; presynaptic; reaction; crisis; receptor; response; social role; stress response; stress; reaction; transmission process

Agonist; Amino Acids; Animal Model; Animal Models and Related Studies; Area; Assay; Attention; Bioassay; Biochemical; Biologic Assays; Biological Assay; Blood Plasma; Brain; CRF receptor type 1; CRF-R1; CRF1 receptor; CRH-1; Cannabinoids, Endogenous; Chemicals; Common Rat Strains; Consultations; Crh1 receptor; Encephalon; Encephalons; Endocannabinoids; Ethanol dependence; Experimental Designs; Glutamates; Immunologic, Radioimmunoassay; Investigation; L-Glutamate; Ligands; Mammals, Primates; Mammals, Rats; Measurement; Methods; Methods and Techniques; Methods, Other; Microdialysis; Molecular; Nervous System, Brain; Neuropeptide Tyrosine; Pilot Projects; Plasma; Primates; RIA; Radioimmunoassay; Rat; Rattus; Reproducibility; Reticuloendothelial System, Serum, Plasma; Sampling; Serum, Plasma; Standardization; Standards; Standards of Weights and Measures; Steroid Compound; Steroids; Techniques; alcohol research; aminoacid; brain tissue; corticotropin-releasing factor receptor 1; design; designing; ethanol addiction; ethanol research; in vivo; interest; model organism; monoamine; neuropeptide Y; pilot study; repository; sample collection; specimen collection; tool

The Administrative Core organizes the administrative functions that will be used by other Cores and Research Components of The Scripps Research Institute Alcohol Research Center (TSRI-ARC). It coordinates all TSRI-ARC activities, and will help support and coordinate the TSRI-ARC with the Center at Large. Functions of the Administrative Core will be to manage scientific oversight by the Executive Committee and coordinate this oversight with the External Advisory Board and Internal Advisory Board. A major role of the Core will be to provide administrative support to the Executive committee, the Cores and assist the External and Internal Advisory Boards in tracking progress toward the achievement of the TSRIARC program goals. It will arrange regular meetings of the Internal Advisory Board, the Executive Committee and the External Advisory Board and monthly meetings of the Participating Investigators. The Administrative Core is organized to ensure proper functioning of the Cores to benefit investigators, their utilization to enhance and expand alcohol research throughout the TSRI-ARC and Center at Large, and to provide review and advisory functions. It will generate periodic reports and disseminate the progress and discoveries to the scientific public via the Education Component. The Administrative Core will coordinate the overall noncompetitive renewals including progress reports and budgets.

Absolute ethanol; African; Alcohol Drinking; Alcohol consumption; Alcohol, Ethyl; Animal Model; Animal Models and Related Studies; Applications Grants; Clinical Research; Clinical Study; Common Rat Strains; Consensus; Data; Dependence; Descriptor; Doctor of Medicine; Doctor of Philosophy; ETOH; East Indian; EtOH drinking; Ethanol; Evaluation; Financial Management; Funding; Future; Genetic; Goals; Grain Alcohol; Grant Proposals; Grants, Applications; Heavy Drinking; Investigators; Lead; M.D.; Macaca mulatta; Mammals, Rats; Methods; Methylcarbinol; Modeling; Molecular; NIH; NRVS-SYS; National Institutes of Health; National Institutes of Health (U.S.); Nervous System; Nervous system structure; Neurologic Body System; Neurologic Organ System; Occupational activity of managing finances; Pb element; Ph.D.; PhD; Phenotype; Pilot Projects; Plant Embryos; Principal Investigator; Process; Programs (PT); Programs [Publication Type]; Progress Reports; Protein Kinase Inhibitors; Qualifying; R01 Mechanism; R01 Program; RPG; Rat; Rate; Rattus; Recommendation; Recruitment Activity; Reports, Progress; Research; Research Activity; Research Grants; Research Personnel; Research Project Grants; Research Projects; Research Projects, R-Series; Researchers; Rest; Rhesus; Rhesus Macaque; Rhesus Monkey; Role; Scientific Evaluation; Scientist; Seeds; System; System, LOINC Axis 4; Time; Trinidad; United States National Institutes of Health; Update; Work; Writing; Zygotes, Plant; adult youth; alcohol ingestion; alcohol intake; alcohol product use; alcohol research; alcohol response; alcohol use; alcoholic beverage consumption; alcoholic drink intake; binge alcohol consumption; binge drinker; binge drinking; bout drinker; design; designing; drink heavily; episodic drinker; episodic drinking; ethanol consumption; ethanol drinking; ethanol ingestion; ethanol intake; ethanol product use; ethanol research; ethanol response; ethanol use; etoh use; excess alcohol consumption; excess alcohol ingestion; excess ethanol ingestion; excessive alcohol consumption; excessive alcohol ingestion; excessive alcohol intake; excessive drinking; excessive ethanol ingestion; extreme drinking; heavy alcohol use; heavy metal Pb; heavy metal lead; innovate; innovation; innovative; member; model organism; neuroadaptation; non-human primate; nonhuman primate; pilot study; pre-clinical; preclinical; programs; protein kinase inhibitor; recruit; response to alcohol; response to ethanol; seed; social role; young adult

The Education Component proposes four interrelated Specific Aims designed to advance NIAAA's mission of translating and disseminating scientific research findings to researchers, healthcare professionals, policy makers, and the general public. Specific Aim 1: Education and Outreach to High Risk Minority and Military Communities. The goal is to develop effective educational and outreach programs for high risk Mexican American, Native American, and military communities in San Diego. We propose to use a lecture/discussion group format which incorporates the latest scientific findings of the Alcohol Research Center at The Scripps Research Institute and demonstrates how those findings inform current diagnostic, treatment, and prevention strategies for alcohol use disorders. The lecture/discussion groups will be specifically tailored to address significant alcohol-related issues in those communities. Treatment professionals, community leaders, parents of adolescents, and spouses of alcohol-affected individuals will be involved in the education programs, rather than alcohol misusing individuals themselves. The programs will be evaluated as to how well they transfer knowledge, improve beliefs, and change behavior about alcohol misuse and use disorders. Specific Aim 2: Integration, Translation, Training, and Education in Alcohol Research. Two seminar series for ARC scientists and trainees are proposed to increase integration of basic science and human studies and translational research between the two domains and provide training in alcohol research and education about alcohol use disorders to undergraduates, psychology and pharmacy graduate students, psychiatric residents, addiction psychiatry fellows, addiction professionals, scientists, and alcohol research trainees. Specific Aim 3. The TSRI ARC High School/College Intern Program. This Aim will promote scientific and healthcare careers in the fields of alcohol addiction and minority health in high school and college students, particularly minority students. Specific Aim 4. Dissemination. The TSRI ARC website will disseminate the scientific findings of the TSRI ARC and information on alcohol use disorders to other alcohol researchers, scientists, healthcare professionals, and the general public. Educational material relevant to high risk minority and military communities will be added and expanded as an adjunct to, and a follow-up for, the in-person educational program in those communities discussed in Specific Aim 1.

PROJECT SUMMARY/ABSTRACT Tobacco use continues to be a major public health problem in the United States with approximately one-third of users becoming dependent and tobacco smoking is the leading avoidable cause of death in the United States. The present proposal focuses on animal studies to investigate neurobiological mechanisms that mediate nicotine addiction. An animal model of increased intravenous self-administration of nicotine to the point of dependence has been developed and will be further refined, validated and compared to acute reinforcement models. Preliminary results show that a CRF-1 antagonist can block both the anxiety-like responses associated with acute nicotine withdrawal and the increase in nicotine responding produced by nicotine deprivation in an extended access model. The overall hypothesis of the present proposal is that enhanced nicotine seeking in extended access (nicotine-dependent) animals is in part produced by an overactivity of extrahypothalamic corticotropin-releasing factor (CRF) stress systems and related stress modulatory agents in specific regions of the basal forebrain (extended amygdala). To test this hypothesis, three specific aims are proposed: 1). To validate an extended dependence model of nicotine self-administration in rats. 2). To explore the neuropharmacological mechanisms within specific neurochemical systems in the anxiety-like state of nicotine withdrawal. 3). To explore the neuropharmacological mechanisms within specific sites within the extended amygdala on nicotine self-administration in dependent rats. Animal models of intravenous self- administration, anxiety-like responses, and brain stimulation reward combined with biochemical studies, and systemic and intracerebral microinjections of specific neuropharmacological agents will be employed. PROJECT NARRATIVE The proposed neurobiological studies will provide key information not only for the etiology of a major component of the motivation to continue to smoke once dependent, help identify those individual differences that may lead to vulnerability to dependence, and provide key targets for future medications development for the treatment of nicotine dependence.

DESCRIPTION (provided by applicant: Cocaine addiction is a chronic relapsing disorder in which subjects episodically administer the drug and ultimately transition from nondependent drug use to the compulsive drug use of addiction. A progressive increase in the frequency and intensity of cocaine use, and a high propensity to relapse after abstinence are two of the major behavioral phenomenon that characterizes the development of cocaine addiction. Despite major advances in understanding the neurobiological mechanisms underlying the transition to cocaine dependence, there are no pharmacological treatments for cocaine dependence. Recently, deep brain stimulation of the subthalamic nucleus has been proposed has a surgical strategy for obsessive-compulsive disorders, but it has never been tested in preclinical models of compulsive drug taking and drug seeking. The subthalamic nucleus, a cerebral structure belonging to the basal ganglia and classically associated with motor control, is critically involved in key cognitive processes that become dysfunctional in subjects with drug addiction. The overall objective of this proposal is to use a new, potentially groundbreaking therapeutic approach for the treatment of cocaine addiction using an innovative neurosurgical approach that has shown remarkable results in other brain and mental disorders, associated with highly relevant animal models of compulsive cocaine intake and relapse to cocaine seeking. Preliminary results show that lesion of the subthalamic nucleus limits the escalation of cocaine intake in dependent rats, and that deep brain stimulation of the subthalamic nucleus decreases the motivation for cocaine in rats. Unknown is whether deep brain stimulation of the STN will reverse the escalation of cocaine intake and prevent relapse to cocaine seeking in dependent rats. The specific objectives of this proposal are to determine whether it is possible to reverse the escalation of cocaine (SpA1), and to prevent drug-, stress-, and cue-induced reinstatement to cocaine seeking (SpA2) using deep brain stimulation in the subthalamic nucleus in cocaine dependent rats. These studies will provide new findings on the role of the subthalamic nucleus in the compulsivity associated with cocaine dependence and may open new avenues for the development of innovative treatments of drug addiction in general. PUBLIC HEALTH RELEVANCE: Despite major advances in understanding the neurobiological mechanisms underlying the transition to drug addiction there are no pharmacological treatments available. The overall objective of this proposal is to use new, potentially groundbreaking therapeutic approaches for cocaine addiction using deep brain stimulation of the subthalamic nucleus, associated with highly relevant animal models of compulsive cocaine intake and relapse to cocaine seeking. These studies will provide new findings on the neurobiological substrates of compulsive cocaine taking and craving, have direct translational implications for drug abuse, and may open new avenues for the development of innovative treatments of drug addiction in general.

DESCRIPTION (provided by applicant): Alcoholism is a chronic relapsing disorder characterized by compulsive use and loss of control over intake. Alcoholism produces significant cost to society in the United States and worldwide. The excessive use of alcohol has long been shown to have detrimental effects on prefrontal cortex function including impairment in decision making, executive function, and memory and learning. In addition, many studies have established that brain stress systems are activated by excessive drinking. However, few studies have explored how chronic alcohol and activation of the brain stress system interacts with the prefrontal cortex to produce cognitive dysfunction and contribute to compulsive alcohol intake. The overall hypothesis of this project is that activation of the brain stress systems [corticotropin releasing factor (CRF) and norepinephrine (NE)] in the prefrontal cortex disrupts cognitive function that exacerbates the powerful motivation for alcohol seeking associated with compulsive use. To address this hypothesis, the present proposal has been designed to (1) To further characterize the time course of development of cognitive dysfunction and compulsive drinking in animal models of excessive drinking. (2) To determine the pattern of changes in the stress systems in the prefrontal cortex in the development of compulsive drinking and (3) To test if chronic inactivation of the stress systems in the prefrontal cortex prevents cognitive deficits and the development of compulsive alcohol drinking. The approach combines neuroanatomical, neuropharmacological, and molecular techniques and the use of innovative animal models of alcohol dependence, such as the escalation-binge and dependence-induced drinking models, combined with very specific measures of compulsive alcohol drinking, working memory and perseverative responding. Understanding the neurobiological mechanisms within the prefrontal cortex that produce cognitive deficits and contribute to the compulsivity of ethanol dependence will provide key information for understanding the individual differences in vulnerability to develop alcoholism and new targets for the treatment and prevention of alcoholism.

The Administrative Core will provide leadership in research, research methodology development, and information dissemination on topics relevant to our overall mission. It coordinates all TSRI-ARC activities, and will help support and coordinate the TSRI-ARC with the Center at Large. Functions of the Administrative Core will be to provide scientific and administrative leadership for the Alcohol Research Center (ARC) by monitoring and enhancing collaborative interactions among the support core components and research components, to facilitate the dissemination of expertise between support core components, pilots and research components by organizing regular TSRI-ARC meetings, seminars, conferences and workshops to stimulate exchange of scientific information among the Center faculty and staff, to promote the sharing of multidisciplinary resources and services among the support core components and investigators, to administer a pilot project program to stimulate new alcohol research programs within the TSRI-ARC at Large, and to disseminate new research knowledge to academic, treatment and lay communities by engaging in community outreach activities designed to improve awareness, prevention and treatment of alcoholism and alcoholic diseases, and supporting regional and national research initiatives aimed at improving prevention and treatment of alcohol use disorders and addiction. It will arrange regular meetings of the Steering Committee and Program Advisory Board and monthly meetings of the Participating Investigators. The Administrative Core is organized to ensure proper functioning of the Center to benefit investigators, their utilization of Cener resources to enhance and expand alcohol research throughout the TSRI-ARC and Center at Large, and to provide review and advisory functions

DESCRIPTION (provided by applicant): The Alcohol Research Center of The Scripps Research Institute (TSRI-ARC) proposes to continue its interdisciplinary program focused on the theme of the central nervous system effects of alcohol. For this renewal application, the TSRI-ARC will be a P60 consisting of 9 components plus an Educational Component. Four core components are proposed: Administrative, Animal Models/Biochemical Measures, Biochemical and Pilot. Five research components are proposed: Cellular Electrophysiology, Cellular Physiology, Neuroendocrinology, Neurochemistry, and Clinical Neurobehavioral. The overall hypothesis of the TSRI-ARC is that with chronic binge drinking, the brain reward systems become tolerant to alcohol while central stress systems become activated and that as a result, the neuroadaptive changes associated with chronic drinking produce an evolving set of neurobehavioral symptoms that include hypohedonia, anxiety, hyperarousal, sleep disturbances, and negative affect, conceptualized as the dark side of addiction. The subhypotheses for the present proposal are: (1) The transition from low levels of drinking (1-2 drinks within 2 hours in humans) to chronic binge drinking (4-5 drinks within 2 hours in humans) is driven by decreased activity of opioid peptide systems and endocannabinoid systems in reward circuits in the frontal cortex, and nucleus accumbens (Specific Aim 1). (2) The transition from binge drinking to dependence is driven by compromised function in the reward systems and recruitment of a dysregulated central stress system, most notably driven by changes in CRF, glutamate, and endocannabinoids in the extended amygdala (Specific Aim 2). (3) A particularly vulnerable local human clinical population has a phenotype of alcoholism that displays this transition from bingeing to dependence in young adulthood, allowing us to translate our findings in animals to humans and humans to animals (Specific Aim 3). We believe the proposed innovative approaches for testing these hypotheses will provide valuable insight into novel approaches for treating and preventing alcoholism in the human population. The TSRI-ARC also supports the Center at Large, which includes: 13 ROIs, 6 UOIs, one T32 NIAAA training grant, two R37s, one R13, one RC1 award. Members of the Center at Large have access to the Cores of the TSRI-ARC, the INIA Cores and the TSRI NIAAA Training Grant in Neuropsychopharmacology. Training and information dissemination to the San Diego community will be effected by the training opportunities of the Center including an NIAAA training grant and the Education Component.`

The Pilot Component The Scripps Research Institute Alcohol Research Center- (TSRI-ARC) will provide a program for conducting pilot studies that advance the research agenda of the TSRI-ARC on neuroadaptive mechanisms associated with the transition from binge drinking to dependence and potentially generate independent grant applications relevant to the focus of the Center at Large. The principal goals of the Pilot Project Program are to enable the TSRI-ARC to explore new directions for innovative research related to the center's goals (Specific Aim 1) and to recruit scientists new to alcohol research into the field, thereby exposing center investigators to fresh perspectives and methods (Specific Aim 2). In general, it is expected that the Pilot projects will provide seed funding to qualified investigators to enable them to gather sufficient preliminary data to attract support for testing a new hypothesis through R01, R21, KO1 or similar mechanisms (or to eliminate a new hypothesis as not worth pursuing). Ultimately the aim of the Pilot Studies component is to provide the TSRI-ARC with a flexible means to develop and explore new research activities or directions and unique scientific opportunities that have the potential to evolve into independently-funded research projects. The proposed pilot studies are integrated into the overall TSRI-ARC program and involve innovative approaches, which will have translational impact across the research components. Emphasis has been placed, in the first two years, on studies that will develop innovative new exciting neurobiological approaches to neurocircuitry targets and on human translational studies for the neuroadapatations associated with excessive drinking and dependence. As demonstrated with the success of our previous Pilot programs, we anticipate that the results of the pilots will help launch innovative lines of research and in parallel new careers in the study of the neurobiology of alcoholism.

The Animal Models/Biochemical Measurement Core of The Scripps Research Institute Alcohol Research Center (TSRI-ARC) will provide a variety of behavioral and bioanalytical services to meet the specific needs of the Center at large. The first goal of the Core is to provide animals engaged in excessive drinking or have a history of excessive drinking using the intermittent access to ethanol drinking (IAE) model or chronic ethanol-induced dependence (CEID) model to TSRI-ARC and Center at Large investigators (Specific Aim 1). In addition, the Core will also supervise all changes in equipment and procedures and any refinement of the current animal models to ensure that standardized procedures are used across all laboratories. The second goal of the Core is to perform biochemical measurements, such as blood alcohol, cortisol/corticosterone, and ACTH levels and brain amino acid and endocannabinoid content, in support of all Center-related projects by establishing state-of-the-art techniques and an efficient, user-friendly website to schedule services, monitor progress, and receive data (Specific Aim 2). Finally, the last goal of the Core is to further characterize the animal models of excessive drinking to be utilized by the TSRI-ARC, including exploring the effect of excessive drinking on the transition to dependence and the effect of excessive drinking on the brain stress system by characterizing the consequence of a history of binge drinking (IAE model) on the brain stress system and the transition to alcohol dependence. Translational dependent measures also used in the Clinical Neurobehavioral Research Component will be employed (Specific Aim 3). The Animal Models/Biochemical Measurement Core will enable Center investigators to enrich the interpretational power of their experiments through investigations of behavioral, neurochemical, neuroendocrine, and pharmacokinetic processes contributing to alcohol dependence.

The ultimate goal of our Section is to understand how cellular and molecular changes produce changes in particular neurocircuits to convey negative emotional states that contribute to the motivation to seek drugs. Currently, we have been investigating the neurochemical and neuroanatomical basis underlying alcohol, heroin, and psychostimulant dependence. Alcohol: We investigated the effects of oxytocin on compulsive alcohol drinking. In this project, rats are trained to lever press for access to alcohol (i.e., operant conditioning). Another lever gives access to water. In this procedure, rats exhibit stable levels of drinking (nondependent) in daily 30 min sessions (about 30 lever presses for alcohol). Then, some of the animals are made dependent on alcohol via chronic, intermittent alcohol vapor exposure (dependent). This results in a dramatic increase in alcohol self-administration in the same period of time (more than 60 lever presses for alcohol in 30 min). Using this model, we observed that oxytocin (a neuropeptide) is capable of substantially reducing compulsive alcohol consumption observed in dependent animals, without disrupting the behavior of nondependent control animals (i.e., the effect is specific to alcohol dependence). This project is highly translational, as we are working to provide data as the basis for a clinical trial of oxytocin in alcohol dependent humans, to be conducted in collaboration with the NIAAA. Heroin: Experimental evidence from human patients, monkeys, and rats indicates that neutral cues can be associated with opioid withdrawal. Presentation of these cues alone can increase operant responding for access to heroin and this constitutes a model of compulsive drug seeking and taking. Here, we developed a rat model of conditioned heroin withdrawal using a heroin self-administration paradigm and naloxone-precipitated withdrawal paired with unique odor cues. We aim to identify the neural circuitry mediating the acquisition and expression of conditioned withdrawal cues. To achieve this we are using: 1. in vivo functional magnetic resonance imaging, and 2. molecular imaging (e.g., immunohistochemistry and in situ hybridization). Additionally, we are further characterizing behavioral responses following presentation of conditioned withdrawal cues in a variety of paradigms (e.g., pain) to understand how these cues affect behavior. Preliminary data indicate a number of cortical and subcortical brain regions to be specifically related to cues that were paired with negative emotional states of opioid withdrawal. Methamphetamine: Methamphetamine is a highly addictive stimulant that has been shown to be substantially detrimental to people suffering from methamphetamine addiction. There is currently no pharmacological treatment for methamphetamine abuse or relapse prevention. This study explores the effects of R-Modafinil, an FDA-approved wake-promoting agent that acts on monoamine transporters, and its analogues on methamphetamine self-administration. In this study, rats were trained to self-administer methamphetamine (0.05 mg/kg) intravenously in 1-h or 6-h sessions. Animals with daily access to the drug for 6 h progressively escalate their intake, whereas animals with access to the drug for 1 h exhibit stable drug intake levels over time. Upon escalated methamphetamine intake, the rats were given intraperitoneal injections of vehicle, modafinil or the DAT-selective analogues, JJC8-016, JJC8-089, and JJC8-091 and tested for methamphetamine self-administration. Thus far, we have data indicating that JJC8-016 and JJC8-091 are capable of reducing methamphetamine self-administration. Cocaine: Cocaine abuse affects approximately 1.7 million individuals nationwide per year and is characterized by patterns of excessive drug seeking and taking, including a preoccupation with obtaining the drug, repetitive seeking and taking of the drug, and a loss of control over drug intake. Compulsive-like cocaine taking and reinstatement of drug seeking (a model of drug relapse) occur in part through neuroadaptations of brain stress systems that mediate negative emotional states implicated in motivational processes required for maintaining the dependent drug state. The goal of the current research is to characterize the role of neuropeptides, including hypocretin/orexin and dynorphin in the mediation of cocaine intake and the reinstatement of drug seeking following extinction. To date, we have shown hypocretin- or dynorphin-receptor antagonism within target extended amygdala brain regions results in attenuation of drug taking and seeking. Future studies are employing adeno-associated viral vectors targeting the hrct or pdyn gene to silence transcription of the peptides in particular brain regions.