Abstract:
This invention describes gene targets for the development of therapeutics to treat drug addiction. Animal models of drug craving and relapse have been developed and used to find gene expression changes in key brain regions implicated in cocaine addiction. The genes whose expression levels are altered serve as pharmacological targets with the purpose of preventing or inhibiting cocaine craving and relapse in human cocaine addicts.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS  
       [0001]     This application is a National Stage of US application number PCT/US02/11094, filed Apr. 4, 2002; said application claims the benefit under 35 USC § 119(e) of U.S. provisional application No. 60/281,440 filed Apr. 4, 2001. The aforementioned applications are incorporated herein by reference for all purposes. 
     
    
     BACKGROUND OF THE INVENTION  
       [0002]     1. Field of the Invention  
         [0003]     The present invention relates to the identification of differentially expressed genes in the brain that are involved in behavior associated with cocaine addiction. More particularly, the present invention relates to methods of identifying and using candidate agents to treat cocaine addiction based upon these genes.  
         [0004]     2. Description of the Related Art  
         [0005]     Drug and alcohol addictions are mental illnesses that exact an enormous social and economic cost from society. Although biomedical research has made tremendous advances in our understanding of how drugs affect the brain, very little of this information has translated into effective treatment strategies. This problem is particularly troublesome for cocaine addiction, where no effective treatments currently exist. Although many cocaine addicts can abstain from drug use for short periods of time, relapse rates at longer periods of abstinence are remarkably high, sometimes exceeding 90% (see Leshner, “Addiction Is a Brain Disease, and It Matters”, Science, Vol. 278, pp. 45-47 (1997)).  
         [0006]     Progress in treating cocaine addiction has been hampered by the failure of animal models to target the primary behavioral disturbance, i.e., the increased propensity for relapse following prolonged periods of abstinence. Recently, the realization of this problem has led investigators to develop new animal models of drug craving in attempts to understand the underlying neurobiological mechanisms that trigger relapse to drug-seeking behavior, and to develop more effective treatment. In these studies, laboratory measures of “cocaine-seeking behavior” provide an objective measure of operant events such as lever-press responses that represent approach behavior analogous to relapse. In these studies, the level of drug-seeking behavior is indicated by the amount of effort (lever-pressing) exerted by animals to self-administer the drug. Importantly, this cocaine-seeking behavior is tested in the absence of drug reinforcement, because the reinforcing and rate-limiting effects of drugs can obscure the true incentive motivational state of the animals. Cocaine-seeking behavior can be measured by the magnitude and persistence of drug-paired lever responding during extinction testing, and by “reinstatement” of this responding following extinction. Either of these measures are thought to reflect the propensity for relapse in humans. Another advantage of these paradigms is that they can be tested during prolonged periods of forced abstinence. In contrast, subjective measures of drug craving in humans can be confounded by the subjective nature of self reports, and contextual differences between laboratory settings and the environment where humans routinely take drugs (see Tiffany et al., “The Development of a Cocaine Craving Questionnaire”, Drug Alcohol Depend., Vol. 34, pp. 19-28 (1993)).  
         [0007]     Generally, there are only three stimuli known to reinstate drug-seeking behavior in animals following extinction of drug-self-administration. These stimuli consist of drug-associated (conditioned) cues, stress and low “priming” doses of the self-administered drug itself (for review, see Self et al., “Relapse to Drug Seeking: Neural and Molecular Mechanisms”, Drug Alcohol Depend., Vol. 51, pp. 49-60 (1998)). Since all three of these stimuli also trigger drug craving in human drug abusers (see Jaffe et al., “Cocaine-Induced Cocaine Craving”, Psychopharmacology, Vol. 97, pp. 59-64 (1989); Robbins et al., “Relationships Among Physiological and Self-Report Responses Produced by Cocaine-Related Cues”, Addictive, Vol. 22, pp. 157-167 (1997); and Sinha et al., “Stress Induced Craving and Stress Response in Cocaine Dependent Individuals”, Pschyopharmacology, Vol. 142, pp. 343-351 (1999) reinstatement of drug-seeking behavior in animals may represent a valid model of human drug craving. One caveat is that human drug addicts rarely, if ever, experience extinction conditions prior to relapse, but the striking concordance of reinstating stimuli in animals, and triggers of craving in humans, suggests that similar neurobiological processes are involved in both reinstatement and craving.  
         [0008]      FIG. 1  depicts some of the primary pathways whereby stress, priming injections of drugs, and drug-associated cues are thought to induce relapse to drug-seeking behavior based on an evolving literature. There is a growing evidence that these stimuli all induce relapse, at least in part, by their ability to elevate dopamine levels in the nucleus accumbens (NAc). Thus, the NAc may be a critical neural substrate for relapse to drug seeking, in addition to its well-characterized role in drug reward. For example, abused drugs which elevate NAc dopamine levels also reinstate cocaine- and heroin-seeking behavior, while abused drugs like barbiturates that do not elevate NAc dopamine levels also fail to reinstate this behavior (reviewed by Self et al., supra). Similarly, infusion of drugs into brain regions where they activate NAc dopamine release reinstates cocaine- and heroin-seeking behavior, where infusion into regions where they do not is without effect.  
         [0009]     Although it has not been clearly resolved, cue- and stress-induced reinstatement of drug-seeking behavior may involve both dopamine-dependent and dopamine-independent neural substrates (reviewed by Self et al., supra). An area of excitatory convergence is the NAc, where excitatory inputs from the prefrontal cortex (PfC), basolateral amygdala (BLA), and subiculum innervate medium spiny neurons receiving dopamine inputs from the ventral tegmental area (VTA). Excitatory neurotransmission in the NAc also has been implicated in reinstatement of cocaine-seeking behavior (see Cornish, et al., “A Role for Nucleus Accumbens Glutamate Transmission in the Relapse to Cocaine-Seeking Behavior”, Neuroscience, Vol. 93, pp. 1359-1367 (1999)). Together, these brain regions all form a complex circuit with primary sites of convergence in both the VTA and NAc of the mesolimbic dopamine system, as depicted in  FIG. 1 .  
         [0010]     These studies highlight new and important information on the neural mechanisms of drug craving and relapse to drug seeking. Given that drug-seeking and drug craving can persist (or increase) despite long periods of abstinence, many current theories suggest that relatively long-term neuroadaptations in limbic brain regions associated with drug-seeking behavior underlie the propensity for relapse in addicted individuals. Most of these theories focus on pharmacological neuroadaptations directly produced by repeated drug exposure, leading to the phenomena of tolerance and sensitization (see Koob et al., “Drug Abuse: Hedonic Homeostatic Dysregulation”, Science, Vol. 278, pp. 52-58 (1997) and Nestler et al., “Molecular and Cellular Basis of Addiction”, Science, Vol. 278, pp. 58-63 (1997)). However, there is little evidence that most neuroadaptations persist during prolonged periods of abstinence (see White et al., “Neuroadaptions Involved in Amphetamine and Cocaine Addiction”, Drug Alc. Dep., Vol. 51, pp. 141-153 (1998)), and, thus, they cannot fully account for the propensity for relapse at these later time points. A major gap in our current knowledge is identifying stable neuroadaptations that underlie persistent drug craving in prolonged abstinence.  
         [0011]     Recently, a behavioral paradigm in rats has been developed that models persistent craving for cocaine during prolonged abstinence. In fact, rats actually show increased levels of cocaine-seeking behavior as abstinence proceeds, a phenomenon also recently reported by Neisewander and colleagues (see Tran-Nguyen et al., “Time-Dependent Changes in Cocaine-Seeking Behavior and Extracellular Dopamine Levels in The Amygdala During Cocaine Withdrawal”, Neuropsychopharmacology, Vol. 19, pp. 48-59 (1998)). In this model, the level of cocaine-seeking behavior progressively increases from 1-6 weeks of forced abstinence from cocaine self-administration. The model is referred to as the “Cocaine Abstinence Effect”, and is thought to reflect time-dependent increases in cocaine craving that lead to increased relapse rates during prolonged abstinence. The model also represents the phenomenon of incentive sensitization, whereby drug-associated stimuli (environmental context, conditioned cues) show enhanced ability to stimulate craving as abstinence proceeds (see Robinson et al., “The Neural Basis of Drug Craving: An Incentive-Sensitization Theory of Addiction”, Brain Res. Rev., Vol. 18, pp. 247-291 (1993)).  
         [0012]     In this model, rats are allowed to acquire intravenous cocaine self-administration on a fixed ratio 1:time-out 15-second schedule of reinforcement for 4 hours/day. Following 12 days of cocaine self-administration, different periods of forced abstinence are imposed whereby animals remain in their home cages, and are not allowed access to the self-administration test chambers. After a given period of abstinence, the rats are returned to the self-administration chambers, and the degree of drug-seeking behavior is measured by the number of non-reinforced responses at the drug-paired lever during extinction testing.  FIG. 2  shows that cocaine-seeking behavior is approximately tripled when rats are returned to the test chambers during the third and sixth week of abstinence, relative to rats returned during their first week of abstinence. Six weeks of abstinence also produces more persistent cocaine-seeking behavior over the first few days of testing. By the sixth day of extinction testing, all three groups have extinguished to similar levels.  
         [0013]      FIG. 3A  shows time-dependent changes in the initial level of cocaine-seeking behavior when rats are first returned to the self-administration test chambers following forced abstinence. Rats tested after 2 and 5 weeks of forced abstinence exhibit 5- to 6-fold greater levels of drug-seeking behavior than at 1 day of abstinence. At the end of extinction testing, rats were tested for cue-induced reinstatement of cocaine-seeking behavior. In this test, cues specifically associated with the 10-second cocaine infusions during self-administration (house light off; lever cue light on; pump noise, vehicle infusion) were presented every 2 minutes for the final hour of the extinction/reinstatement test session.  FIG. 3B  shows that the cocaine abstinence effect is still evident following extinction testing, but only in the group tested during their sixth week of forced abstinence. Thus, cues specifically associated with cocaine infusions during self-administration induced greater reinstatement of responding at 6 weeks of abstinence than at 1 week of abstinence. Moreover, extinction training failed to completely reverse the Cocaine Abstinence Effect in this 6-week group, although cue-induced reinstatement at 3 weeks abstinence failed to differ as in extinction testing.  
         [0014]     The Cocaine Abstinence Effect suggests that the incentive motivational effects of the drug-paired environment (extinction), and cocaine-associated cues (reinstatement), gain motivational salience with prolonged abstinence from cocaine. In contrast, pharmacological models of drug addiction and dependence suggest that drug craving would be maximal during early abstinence periods, when withdrawal symptoms also are maximal, and diminish as withdrawal effects wane over time (see Koob et al., supra).  
         [0015]     The model closely parallels a similar effect of prolonged abstinence from chronic alcohol consumption known as the “alcohol deprivation effect” (see Sinclair, “The Alcohol-Deprivation Effect. Influence of various factors.”, Quarterly Journal of Studies on Alcohol, Vol. 33, pp. 769-782 (1972)), although it differs by measuring drug-seeking behavior rather than drug intake. This feature represents an important advantage over models of drug intake, because acute effects of drugs following abstinence could obscure certain biochemical measures that correlate with time-dependent increases in cocaine-seeking, and the response rate-limiting effects of drugs could alter behavioral measures of drug seeking. As cited above, at least one other group has published the phenomenon of time-dependent increases in cocaine seeking using similar (2 and 4 weeks) periods of forced abstinence (see Tran-Nguyen et al., supra). This study found that the behavioral effects also were associated with increased basal dopamine levels in the central nucleus (CeA) of the amygdala, and greater increases in dopamine release when animals were first returned to the self-administration chambers during extinction testing.  
         [0016]     The “Cocaine Abstinence Effect” animal model is particularly useful in understanding the underlying biochemical neuroadaptations that trigger relapse to drug-seeking behavior. Accordingly, the use of this model to identify changes in gene expression that coincide with time-dependent increases in cocaine-seeking behavior and extinction training in rats, would aid in identifying potential therapeutic targets and therapeutic agents for use in treating cocaine addiction.  
       SUMMARY OF THE INVENTION  
       [0017]     The present invention is based on the identification of genes found in particular brain regions of rats that are modulated by behavior associated with cocaine addiction and extinction training. The genes have been identified by using a behavior animal model of cocaine addiction combined with oligonucleotide array profiling techniques. In particular, the present invention is directed to methods for inhibiting behavior associated with cocaine addiction in a subject such as a mammal suffering from cocaine addiction, and methods for identifying candidate agents useful in inhibiting behavior associated with cocaine addiction, using these genes.  
         [0018]     In some embodiments, the invention provides methods for inhibiting addiction-related behavior in a subject suffering from cocaine addiction. These methods involve administering to the subject a therapeutically effective amount of a therapeutic agent which has the ability to modulate the level of activity of a polypeptide encoded by at least one gene identified in one or more of Tables 1-15. The activity of the polypeptide can be modulated by, for example, increasing or decreasing the level of expression of a gene that encodes the polypeptide, the level at which a transcript is translated or maintained in a cell, or by increasing or decreasing the enzymatic activity, binding ability, or other property of the polypeptide itself.  
         [0019]     The invention also provides methods of inhibiting addiction-related behavior in a subject suffering from cocaine addiction that involve administering to the subject a therapeutically effective amount of a therapeutic agent which has the ability to decrease transcription/translation of, or decrease the activity of a protein encoded by, at least one gene that encodes a polypeptide selected from the group consisting of hypertension-regulated vascular factor, myelin-associated basic protein, PB cadherin, calcitonin receptor, melanocortin 4 receptor, ALK-7 kinase, and retroposon.  
         [0020]     Also provided are methods of inhibiting addiction-related behavior in a subject suffering from cocaine addiction that involve administering to the subject a therapeutically effective amount of an agonist that activates a protein selected from the group consisting of GABA-B receptor subunit gb2, cell adhesion-like molecule,  bos taurus -like neuronal axonal protein, a polypeptide similar to mouse chemokine-like factor, FRA-2, a protein similar to human oxygen-regulated protein, a protein similar to mouse mrg1 protein, pentraxin, malic enzyme, olfactomedin-related protein, arc-growth factor, protein tyrosine phosphatase, krox, neuritin, microtubule-associated protein 2d, and CB1 cannabinoid receptor.  
         [0021]     Another aspect of the invention provides methods for identifying an agent to be tested for an ability to prevent or inhibit cocaine addiction-related behavior. These methods can involve: a) combining in a reaction mixture a candidate agent with a protein encoded by a gene identified in Tables 1-15; and b) determining whether the candidate agent binds to and/or modulates activity of the protein.  
         [0022]     In some embodiments, these methods can further involve adding to the reaction mixture a competitor molecule that competes with binding of the candidate agent to the protein, either prior to or subsequent to combining the protein with the candidate agent.  
         [0023]     In other embodiments, the methods further involve: c) administering the candidate agent identified in b) to a cocaine-addicted subject or brain cells of a cocaine-addicted subject, wherein the cocaine-addicted subject is undergoing withdrawal; and d) determining a level of expression of at least one gene identified in Tables 1-15 in brain cells of the cocaine-addicted subject. The level of expression is compared to that observed in brain cells of a cocaine-addicted subject to which the candidate agent is not administered, wherein a change in expression level is indicative of the candidate having efficacy in preventing or inhibiting cocaine addiction-related behavior.  
         [0024]     Still other embodiments involve: c) administering the candidate agent identified in b) to a cocaine-addicted subject that is undergoing withdrawal; and d) determining whether the withdrawal symptoms exhibited by the subject are reduced upon administration of the candidate agent.  
         [0025]     Also provided by the invention are methods for identifying an agent to be tested for an ability to prevent or inhibit addiction related behavior. These methods involve: a) exposing a cocaine-addicted subject or brain cells of a cocaine-addicted subject to a candidate agent, wherein the cocaine-addicted subject is undergoing withdrawal; b) determining a level of expression of at least one gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject, wherein the at least one gene is identified in Tables 1-15; and c) comparing the level of expression of the gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject in the presence of the candidate agent with the level of expression of the gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject in the absence of the candidate agent. A reversal in the level of expression of the gene in cocaine-addicted subject or brain cells of the cocaine addicted subject in the presence of the candidate agent relative to the level of expression of the gene in the absence of the candidate agent indicates that the candidate agent is an agent to be tested for the ability to prevent or inhibit addiction related behavior.  
         [0026]     The invention also provides methods for identifying an agent to be tested for an ability to prevent or inhibit cocaine addiction-related behavior. These methods involve: 
        a) contacting a brain tissue sample from each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     b) detecting a level of expression of at least one gene in both tissue samples, wherein the gene encodes a polypeptide selected from the group consisting of hypertension-regulated vascular factor, myelin-associated basic protein, PB cadherin, calcitonin receptor, melanocortin 4 receptor, ALK-7 kinase and retroposon.     c) subtracting the level of expression of the gene in the sample obtained from the cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the subject having cocaine addiction-related behavior to provide a first value;     d) administering a candidate agent to each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     e) detecting a level of expression of at least one gene in both tissue samples obtained from the subjects treated with the candidate agent;     f) subtracting the level of expression of the at least one gene in the sample obtained from the treated cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the treated subject having the cocaine addiction-related behavior to provide a second value; and     g) comparing the second value with the first value wherein a decreased second value relative to the first value is indicative of an agent to be tested for an ability to prevent or inhibit cocaine addiction-related behavior.        
 
         [0034]     In some embodiments, the invention provides methods for identifying agents to be tested for an ability to prevent or inhibit cocaine addiction-related behavior that involve: 
        a) obtaining a brain tissue sample from each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     b) detecting a level of expression of at least one gene in both tissue samples, wherein the gene encodes a polypeptide selected from the group consisting of GABA-B receptor subunit gb2, cell adhesion-like molecule,  bos taurus -like neuronal axonal protein, similar to mouse chemokine-like factor, FRA-2, a polypeptide similar to human oxygen-regulated protein, a polypeptide similar to mouse mrg1 protein, pentraxin, malic enzyme, olfactomedin-related protein, arc-growth factor enriched in dendrites, protein tyrosine phosphatase, krox, neuritin, microtubule-associated protein 2d and CB1 cannabinoid receptor;     c) subtracting the level of expression of the gene in the sample obtained from the cocaine addiction-free subject from the level of expression of the gene of the sample obtained from the subject having cocaine addiction-related behavior to provide a first value;     d) administering a candidate agent to each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     e) detecting a level of expression of the gene in both tissue samples obtained from the subjects treated with the candidate agent;     f) subtracting the level of expression of the gene in the sample obtained from the treated cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the treated subject having the cocaine addiction-related behavior to provide a second value; and     g) comparing the second value with the first value wherein an increased second value relative to the first value is indicative of an agent to be tested for an abilty to prevent or inhibit cocaine addiction related behavior.        
 
     
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0042]      FIG. 1 . Diagrammatic representation of the primary pathways through which stress, drugs of abuse and drug-associated conditioned stimuli are hypothesized to trigger drug craving and relapse to drug-seeking. Stress and conditioned stimuli can activate excitatory glutamatergic projections to the VTA from the PfC, amygdala (Amyg) and hippocampus (Hipp), while priming injections of drugs directly stimulate dopamine (DA) release in the NAc. In this sense, dopamine release in the NAc may be a may be a final common trigger of drug craving by all three stimuli. At the level of NAc neurons, dopamine from the VTA modulates direct excitatory signals from the PfC, Amyg and Hipp where complex spatio-temporal integration of relapse-related information occurs. Studies showing involvement of these brain regions in reinstatement of drug-seeking suggest that long-term changes in gene expression in these regions would alter the functionality of this circuitry, and could produce profound changes in reactivity to stimuli that trigger drug craving and relapse to drug-seeking (adapted from Self et al., supra).  
         [0043]      FIG. 2 . Time-dependent increases in drug-seeking behavior during forced abstinence from cocaine self-administration. Groups of rats (ns=8-25) were balanced such that each group averaged similar levels of cocaine intake on the last 3 days of self-administration testing (1.0 mg/kg/infusion in 4 hour test sessions during the dark cycle). Following different periods of forced abstinence rats were returned to the drug-paired environment, and non-reinforced responding at the drug-paired lever was measured during 6 daily 4-hour extinction tests. Rats tested during the third and sixth week of forced abstinence showed significantly greater levels of drug-seeking behavior during the first 2 extinction tests than rats tested during the first week of forced abstinence (*P&lt;0.05; Fisher&#39;s LSD).  
         [0044]      FIG. 3 . The Cocaine Abstinence Effect is evident at both the beginning (A) and end (B) of extinction testing. Selective responding at the drug-paired, rather than inactive, lever reflects the level of effort exerted by animals to self-administer cocaine (i.e., drug-seeking behavior). The left panel (A) depicts non-reinforced responding during the first hour of the initial 4-hour extinction test in groups of animals with forced abstinence ranging from 1 day to 5 weeks. The initial level of spontaneous drug-seeking behavior more than tripled at 2 and 5 weeks of forced abstinence when compared to rats tested after 1 day of forced abstinence (***P≦0.001). Following extinction testing, the ability of cues associated with cocaine infusions (house light off; lever light on; pump noise, vehicle infusion) to induce relapse to drug-seeking behavior was measured (B). The cues were non-contingently delivered for 10 seconds every 2 minutes for 1 hour immediately following the final extinction test. The level of drug-seeking behavior during cue-induced relapse doubled at 6 weeks when compared to 1 week of forced abstinence (**P&lt;0.01; Fisher&#39;s LSD; 3-4 non-responders/group were not included in relapse analysis). Note that baseline response rates in the 1-hour period preceding cue exposure were similar for all 3 groups of rats (mean group responses ranged from 5.4-9.2).  
         [0045]      FIG. 4 . Effects of extinction training on withdrawal-induced changes in gene expression following 1 week abstinence from 12 days (4 hours/day at 1.0 mg/kg/injection) of cocaine self-administration. Example GeneChip profiles of mRNAs from NAc shell tissue are shown for 2 genes differentially regulated during early withdrawal by extinction training. Expression of the retroviral derived rat brain retroposon gene is elevated 88% during withdrawal from cocaine self-administration, but decreased 49% in animals that underwent 4 hours/day of extinction training, when compared to control values (see Table 1). The CB1 cannabinoid receptor is reduced 53% during withdrawal from cocaine self-administration, but is normalized (19% increase relative to control values) in animals that experienced extinction training during withdrawal. The top row of highlighted boxes in each array contains several different oligonucleotide sequences (25 bases/each) spanning the target sequence, while the bottom row contains a 1 base mismatch in the same sequences.  
         [0046]      FIG. 5 . Time-course and overall experimental strategy to identify changes in gene expression produced by cocaine self-administration (SA) abstinence and extinction. Arrows denote the time of sacrifice and dissection of the NAc shell for analysis with gene expression profiling. Group I remained in their home cages during 1 week of abstinence. Groups II and IV underwent 1 week of extinction training 1 week prior to sacrifice. Not shown are three groups that simultaneously underwent saline self-administration and were sacrificed along with Groups I, II and IV.  
         [0047]      FIG. 6 . Diagrammatic representation of tissue punches of limbic brain regions collected from animals during 1 week abstinence from cocaine self-administration for oligonucleotide array analysis. A “half-moon” outer punch of NAc shell was collected with a 12-gauge tissue punch. Each punch was taken from chilled brain slices immediately following sacrifice. The anatomical plates illustrate the posterior side of each 1.2-1.5 mm thick brain slice. For the current study, only the NAc shell was used. Other brain regions shown were also dissected but will be used for later studies.  
         [0048]      FIG. 7 . GABA-B receptor subunit gb2 protein levels are increased by extinction training in the NAc shell as measured by Western Blot. Values are expressed as a percentage of the mean of the control group.  
         [0049]      FIGS. 8-10 . Cannabinoid receptor CB1 protein levels are increased by cocaine withdrawal in the NAc shell as measured by Western Blot. Three different bands specific for CB1 were detected and quantitated separately:  FIG. 8 , 70 kDa glycosylated species;  FIG. 9 , upper 50 kDa nonglycosylated species; and  FIG. 10 , lower 50 kDAa glycosylated species. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0050]     All patent applications, patents and literature references cited herein are hereby incorporated by reference in their entirety.  
         [0051]     The present invention relates to the identification of genes that are up- or down-regulated in particular regions of the brain of rats undergoing cocaine withdrawal compared with rats that are free from cocaine addiction (control) as shown below (see Tables 1-16.  
         [0052]     As used herein, the term “up-regulated” with respect to these genes means that the expression of these genes is higher in rats undergoing cocaine withdrawal compared with rats that are free from cocaine addition. Such up-regulation refers to at least about a two-fold change.  
         [0053]     As used herein, the term “down-regulated” with respect to these genes means that the expression of these genes is lower in cocaine-addicted rats undergoing withdrawal compared with rats that are free from cocaine addiction. Such down-regulation refers to at least about a two-fold changed.  
         [0054]     Importantly, as shown in Table 1 the up- or down-regulation of many of these genes observed in the brain tissue of cocaine-addicted rats undergoing withdrawal is reversed upon subjecting these rats to extinction training. These results indicate a causal relationship between extinction-induced neuroadaptations in these genes and the propensity for behavior associated with cocaine addiction, particularly cocaine-seeking behavior. Accordingly, these differentially expressed genes can form the basis for novel agents useful in the treatment of cocaine addiction and in reducing, inhibiting or preventing addiction-related behavior in individuals suffering from cocaine addiction. In addition, these differentially expressed genes can be utilized to identify agents that inhibit or prevent behavior associated with cocaine addiction. Gene expression is typically assessed about 1-2 weeks after withdrawal.  
         [0055]     The complete sequences of the genes listed in Tables 1-15 are available from GenBank database using the assigned accession numbers (as in Table 1) or part of the probe set identification numbers which indicate the accession numbers of the genes. For example, Probe set identification number “rc_AA875032_at listed in Table 3 corresponds to GenBank Accession No. AA875032. The sequences of these genes in GenBank, and their probe identification and accession numbers are expressly incorporated herein by reference.  
         [0056]     The brain regions where these genes are differentially expressed include the nucleus accumbens shell (Nac shell), the nucleus accumbens core (Nac core), the central nucleus of the amygdala (CeA), the ventral tegmental area (VTA) and the medial prefrontal cortex (mPFC). Evidence linking behavior associated with cocaine addiction to the aforementioned brain regions further support the involvement of the aforementioned genes expressed in these brain regions in such behavior. As stated above, although it has not been clearly resolved, cue- and stress-induced reinstatement of drug-seeking behavior may involve both dopamine-dependent and dopamine-independent neural substrates (reviewed by Self et al., supra). However, the basolateral amygdala (BLA), as well the CeA and related extended amygdala structures recently have been implicated in cue-and stress-induced reinstatement of drug-seeking behavior (see Meil et al, “Lesions of the Basolateral Amygdala a Bolish of the Ability of Drug Associated Cues to Reinstate Responding During Withdrawal from Self-Administered Cocaine”, Behav., Brain Res., Vol. 87, pp. 139-148 (1997); and Erb et al., “A Role for the Bed Nucleus of the Stria Terminalis, but Not the Amygdala, in the Effects of Corticotroopin-Releasing Factor on Stress-Induced Reinstatement of Cocaine Seeking”, J. Neurosci., Vol. 19, pp. C1-C6 (1999)). The CeA sends a direct excitatory projection to VTA neurons (see Gonzales et al., “Amydalonigral Pathway: An Anterograde Study in the Rat with Phaseolus Vulgaris Leucoagglutinin”, J. Comp. Neurol., Vol. 297, pp. 182-200 (1990); and Wallace et al., “Organization of Amygdaloid Projections to Brainstem Dopaminergic, Noradrenergic, and Adrenergic Cell Groups in the Rat”, Brain Res. Bull., Vol. 28, pp. 447-454 (1992)), which could mediate dopamine release in response to cues and stress. Other brain regions involved in relapse may include the PfC, where excitatory projections to dopamine neurons in the VTA activate dopamine release in the NAc (see Moghaddam, “Stress Preferentially Increases Extraneuronal Levels of Excitatory Amino Acids in the Prefrontal Cortex: Comparison to Hippocampus and Basal Ganglia”, J. Neurochem., Vol. 60, pp. 1650-1657 (1993); Taber, Das and Fibiger, “Cortical Regulation of Subcortical Dopamine Release: Mediation Via the Ventral Tegmental Area”, J. Neurochem., Vol. 65, pp. 1407-1410 (1995); and Karreman et al., “The Prefrontal Cortex Regulates the Basal Release of Dopamine in the Limbic Striatum: An Effect Mediated by Ventral Tegmental Area”, J. Neurochem., Vol. 66, pp. 589-598 (1996)). Similarly, recent studies have found that electrical stimulation of hippocampal-subiculuar outputs elevates dopamine levels in the NAc via excitatory inputs to the VTA (see Legault et al., “Chemical Stimulation of the Ventral Hippocampus Elevates Nucleus Accumbens Dopamine by Activating Dopaminergic Neurons of the Ventral Tegmental Area”, J. Neurosci., Vol. 20, pp. 1635-1642 (2000)), and also reinstates cocaine-seeking behavior (see Vorel et al., “Electrical Stimulation of Ventral Subiculum Induced Relapse to Cocaine Self-Administration”, Soc. Neurosci. Abstr., p. 2170 (1998). Another area of excitatory convergence is the NAc, where excitatory inputs from these the PfC, B1A and subiculum innervate medium spiny neurons receiving dopamine inputs from the VTA. Excitatory neurotransmission in the NAc also has been implicated in reinstatement of cocaine-seeking behavior (see Cornish et al., supra). Together, these brain regions all form a complex circuit with primary sites of convergence in both the VTA and NAc of the mesolimbic dopamine system, as depicted in  FIG. 1 .  
         [0057]     Any selection of at least one of the genes listed in Tables 1-15 can be utilized as a therapeutic target for inhibiting or preventing behavior associated with cocaine addiction. Preferably at least one of the genes is identified in Tables 1, 5, 8, 11 and 14, and more preferably at least one gene is identified in Table 1. In particularly useful embodiments, a plurality of these genes, i.e. two or more, can be selected and their expression monitored simultaneously to provide expression profiles for use in various aspects. For example, expression profiles of these genes can provide valuable molecular tools for rapidly identifying agents that alter these expression profiles. Particularly preferred genes from Tables 1-15 that are useful as therapeutic targets include those listed in Table 16.  
         [0058]     In one aspect, methods of treating addiction-related behavior in a subject, e.g., a human or animal, suffering from cocaine addiction are provided which involve preventing or inhibiting cocaine-addiction related behavior utilizing various therapeutics that modulate the transcription/translation of these differentially expressed genes or that modulate the activity of proteins encoded by these genes. As used herein, cocaine refers to cocaine itself and derivatives of cocaine, e.g., crack. As used herein the term “addiction-related behavior” refers to behavior resulting from cocaine use and is characterized by apparent total dependency on cocaine. Symptomatic of such behavior is (i) overwhelming involvement with the use of cocaine; (ii) the securing of its supply; and (iii) a high probability of relapse following withdrawal. For example, in cocaine users addiction-related behavior typically includes behavior associated with three stages of drug effects. In the first stage, acute intoxication, “binge”, is euphoric, marked by decreased anxiety, enhanced self-confidence and sexual appetite. In the second stage, the “crash” replaces the euphoric feeling with anxiety, fatigue, irritability and depression. The third stage, “anhedonia” is a time of limited ability to experience pleasure from normal activities and of craving for the euphoric effects of cocaine. In particularly useful embodiments, the cocaine-addiction related behavior is cocaine seeking. As used herein, cocaine seeking which is a behavior measured in cocaine-addicted animals such as rats is assumed to be analogous to the behavior, cocaine craving, that is observed in humans.  
         [0059]     Examples of suitable therapeutic agents for inhibiting or preventing cocaine addiction-related behavior include, but are not limited to, antisense sequences, ribozymes, double-stranded RNAs, small inhibitory RNA (siRNA), agonists and antagonists as described in detail below.  
         [0060]     As used herein, the term “antisense” refers to nucleotide sequences that are complementary to a portion of an RNA expression product of at least one of the disclosed genes. “Complementary” nucleotide sequences refer to nucleotide sequences that are capable of base-pairing according to the standard Watson-Crick complementarity rules. That is, purines will base-pair with pyrimidine to form combinations of guanine:cytosine and adenine:thymine in the case of DNA, or adenine:uracil in the case of RNA. Other less common bases, e.g., inosine, 5-methylcytosine, 6-methyladenine, hypoxanthine and others may be included in the hybridizing sequences and will not interfere with pairing.  
         [0061]     When introduced into a host cell, antisense nucleotide sequences specifically hybridize with the cellular mRNA and/or genomic DNA corresponding to the gene(s) so as to inhibit expression of the encoded protein, e.g., by inhibiting transcription and/or translation within the cell.  
         [0062]     The isolated nucleic acid molecule comprising the antisense nucleotide sequence can be delivered, e.g., as an expression vector, which when transcribed in the cell, produces RNA which is complementary to at least a unique portion of the encoded mRNA of the gene(s). Alternatively, the isolated nucleic acid molecule comprising the antisense nucleotide sequence is an oligonucleotide probe which is prepared ex vivo and, which, when introduced into the cell, results in inhibiting expression of the encoded protein by hybridizing with the mRNA and/or genomic sequences of the gene(s).  
         [0063]     The oligonucleotide can include artificial internucleotide linkages which render the antisense molecule resistant to exonucleases and endonucleases, and thus are stable in the cell. Examples of modified nucleic acid molecules for use as antisense nucleotide sequences are phosphoramidate, phosporothioate and methylphosphonate analogs of DNA as described, e.g., in U.S. Pat. Nos. 5,176,996; 5,264,564; and 5,256,775. General approaches to preparing oligomers useful in antisense therapy are described, e.g., in Van der Krol., BioTechniques 6:958-976, 1988; and Stein et al., Cancer Res. 48:2659-2668, 1988.  
         [0064]     Typical antisense approaches, involve the preparation of oligonucleotides, either DNA or RNA, that are complementary to the encoded mRNA of the gene. The antisense oligonucleotides will hybridize to the encoded mRNA of the gene and prevent translation. The capacity of the antisense nucleotide sequence to hybridize with the desired gene will depend on the degree of complementarity and the length of the antisense nucleotide sequence. Typically, as the length of the hybridizing nucleic acid increases, the more base mismatches with an RNA it may contain and still form a stable duplex or triplex. One skilled in the art can determine a tolerable degree of mismatch by use of conventional procedures to determine the melting point of the hybridized complexes.  
         [0065]     Antisense oligonucleotides are preferably designed to be complementary to the 5′ end of the mRNA, e.g., the 5′untranslated sequence up to and including the regions complementary to the mRNA initiation site, i.e., AUG. However, oligonucleotide sequences that are complementary to the 3′ untranslated sequence of mRNA have also been shown to be effective at inhibiting translation of mRNAs as described e.g., in Wagner, Nature 372:333, 1994. While antisense oligonucleotides can be designed to be complementary to the mRNA coding regions, such oligonucleotides are less efficient inhibitors of translation.  
         [0066]     Regardless of the mRNA region to which they hybridize, antisense oligonucleotides are generally from about 15 to about 25 nucleotides in length.  
         [0067]     The antisense nucleotide can also comprise at least one modified base moiety, e.g., 3-methylcytosine, 5,-methylcytosine, 7-methylguanine, 5-fluorouracil, 5-bromouracil, and may also comprise at least one modified sugar moiety, e.g., rabinose, hexose, 2-fluorarabinose, and xylulose.  
         [0068]     In another embodiment, the antisense nucleotide sequence is an alpha-anomeric nucleotide sequence. An alpha-anomeric nucleotide sequence forms specific double stranded hybrids with complementary RNA, in which, contrary to the usual beta-units, the strands run parallel to each other as described e.g., in Gautier et al., Nucl. Acids. Res. 15:6625-6641, 1987.  
         [0069]     Antisense nucleotides can be delivered to cells which express the described genes in vivo by various techniques, e.g., injection directly into the prostate tissue site, entrapping the antisense nucleotide in a liposome, by administering modified antisense nucleotides which are targeted to the prostate cells by linking the antisense nucleotides to peptides or antibodies that specifically bind receptors or antigens expressed on the cell surface.  
         [0070]     However, with the above-mentioned delivery methods, it may be difficult to attain intracellular concentrations sufficient to inhibit translation of endogenous mRNA. Accordingly, in a preferred embodiment, the nucleic acid comprising an antisense nucleotide sequence is placed under the transcriptional control of a promoter, i.e., a DNA sequence which is required to initiate transcription of the specific genes, to form an expression construct. The use of such a construct to transfect cells results in the transcription of sufficient amounts of single stranded RNAs to hybridize with the endogenous mRNAs of the described genes, thereby inhibiting translation of the encoded mRNA of the gene. For example, a vector can be introduced in vivo such that it is taken up by a cell and directs the transcription of the antisense nucleotide sequence. Such vectors can be constructed by standard recombinant technology methods. Typical expression vectors include bacterial plasmids or phage, such as those of the pUC or Bluescript.™ plasmid series, or viral vectors such as adenovirus, adeno-associated virus, herpes virus, vaccinia virus and retrovirus adapted for use in eukaryotic cells. Expression of the antisense nucleotide sequence can be achieved by any promoter known in the art to act in mammalian cells. Examples of such promoters include, but are not limited to, the promoter contained in the 3′ long terminal repeat of Rous sarcoma virsu as described, e.g., in Yamamoto et al., Cell 22: 787-797, 1980; the herpes thymidine kinase promoter as described e.g., in Wagner et al., Proc. Natl. Acad. Sci. U.S.A. 78:1441-1445, 1981; the SV40 early promoter region as described e.g., in Bernoist and Chambon, Nature 290:304-310, 1981; and the regulatory sequences of the metallothionein gene as described, e.g., in Brinster et al., Nature 296:39-42, 1982.  
         [0071]     Ribozymes are RNA molecules that specifically cleave other single-stranded RNA in a manner similar to DNA restriction endonucleases. By modifying the nucleotide sequences encoding the RNAs, ribozymes can be synthesized to recognize specific nucleotide sequences in a molecule and cleave it as described, e.g., in Cech, J. Amer. Med. Assn. 260:3030, 1988. Accordingly, only mRNAs with specific sequences are cleaved and inactivated.  
         [0072]     Two basic types of ribozymes include the “hammerhead”-type as described for example in Rossie et al. Pharmac. Ther. 50:245-254, 1991; and the hairpin ribozyme as described, e.g., in Hampel et al, Nucl. Acids Res. 18:299-304, 1999 and U.S. Pat. No. 5,254,678. Intracellular expression of hammerhead and hairpin ribozymes targeted to mRNA corresponding to at least one of the disclosed genes can be utilized to inhibit protein encoded by the gene.  
         [0073]     Ribozymes can either be delivered directly to cells, in the form of RNA oligonucleotides incorporating ribozyme sequences, or introduced into the cell as an expression vector encoding the desired ribozymal RNA. Ribozyme sequences can be modified in essentially the same manner as described for antisense nucleotides, e.g., the ribozyme sequence can comprise a modified base moiety.  
         [0074]     Double-stranded RNA, i.e., sense-antisense RNA, corresponding to at least one of the disclosed genes, can also be utilized to interfere with expression of at least one of the disclosed genes. Interference with the function and expression of endogenous genes by double-stranded RNA has been shown in various organisms such as  C. elegans  as described, e.g., in Fire et al., Nature 391:806-811, 1998; drosophilia as described, e.g., in Kennerdell et al., Cell 95(7):1017-26, 1998; and mouse embryos as described, e.g., in Wianni et al., Nat. Cell Biol. 2(2):70-5, 2000. Such double-stranded RNA can be synthesized by in vitro transcription of single-stranded RNA read from both directions of a template and in vitro annealing of sense and antisense RNA strands. Double-stranded RNA can also be synthesized from a cDNA vector construct in which the gene of interest is cloned in opposing orientations separated by an inverted repeat. Following cell transfection, the RNA is transcribed and the complementary strands reanneal. Double-stranded RNA corresponding to at least one of the disclosed genes could be introduced into a prostate cell by cell transfection of a construct such as that described above.  
         [0075]     The term “antagonist” refers to a molecule which when bound to the protein encoded by the gene inhibits its activity. Antagonists can include, but are not limited to, peptides, proteins, carbohydrates and small molecules. In a particularly useful embodiment, the antagonist is an antibody specific for the protein expressed by the at least one gene.  
         [0076]     The term “agonist” as used herein refers to any natural or synthetic molecule which, when bound to the expressed protein, increases or prolong the duration of the effect of the protein. Agonists can include proteins, nucleic acids, carbohydrates or any other molecules that bind to and modulate the effect of the protein.  
         [0077]     In one embodiment, a method of inhibiting addiction-related behavior in a subject suffering from cocaine addiction is provided which comprises administering to the subject a therapeutically effective amount of a therapeutic agent which has the ability to modulate the transcription/translation of at least one gene or the activity of a protein encoded by the genes, wherein the at least one gene is identified in Tables 1, 2 and 4-15. In the case where the therapeutic agent is an antisense sequence, an isolated nucleic acid molecule encoding a ribozyme, or a double stranded RNA, such an agent modulates the transcription/translation of the gene. In the case wherein the therapeutic agent is an antagonist or agonist, such an agent modulates the activity of a protein encoded by the gene.  
         [0078]     As used herein, the term “isolated” nucleic acid molecule means that the nucleic acid molecule is removed from its original environment (e.g., the natural environment if it is naturally occurring). For example, a naturallly occurring nucleic acid molecule is not isolated, but the same nucleic acid molecule, separated from some or all of the coexisting materials in the natural system, is isolated, even if subsequently reintroduced into the natural system. Such nucleic acid molecules could be part of a vector or part of a composition and still be isolated, in that such vector or composition is not part of its natural environment.  
         [0079]     As used herein, the term “modulate” with respect to transcription/translation refers to the up-or down-regulation of transcription/translation of the gene, i.e., that is “modulate” includes either an increase or a decrease in expression of the at least one gene. The direction of modulation affected by the therapeutic agent depends on which gene is being modulated. For example, the calcitonin receptor gene is upregulated in the Nac Shell of cocaine-addicted rats during cocaine withdrawal. Accordingly, an antisense sequence, a ribozyme, or a double stranded RNA modulates expression of the calcitonin gene by blocking the “up-regulation” of expression of this gene or reversing or “down-regulating” the expression of this gene.  
         [0080]     As used herein, the term “modulate” with respect to activity of a protein encoded by the gene, refers to an alteration, i.e., increase or decrease, in the activity of a protein encoded by the gene. For example, the gene encoding malic enzyme is down-regulated in Nac Shell of cocaine-addicted rats during cocaine withdrawal. Accordingly, an agonist that would increase the activity of the malic enzyme can aid in inhibiting addiction-related behavior.  
         [0081]     In a preferred embodiment of the method for inhibiting or preventing cocaine-addiction related behavior, the at least one gene identified in Table 1 encodes a polypeptide selected from the group consisting of GABA-B receptor subunit gb2, myelin-associated basic protein, calcitonin receptor,  Bos taurus -like neuronal axonal protein, FRA-2, a polypeptide similar to human oxygen-regulated protein, a polypeptide similar to mouse mrg 1 protein, pentraxin, olfactomedin-related protein, arc-growth factor (enriched in dendrites), protein tyrosine phosphatase, melanocortin 4 receptor, ALK-7 kinase, neuritin and CB1 cannabinoid receptor. More preferably, the at least one gene identified in Table 1 encodes GABA-B receptor subunit gb2, FRA-2 and CB1 cannabinoid receptor. In some embodiments of the method for inhibiting or preventing cocaine addiction-related behavior, the at least one gene identified in Table 1 does not encode melanocortin 4 receptor.  
         [0082]     In another preferred embodiment of the method for inhibiting or preventing cocaine addiction-related behavior, the at least one gene is identified in Table 2.  
         [0083]     In another preferred embodiment of the above method, the at least one gene is identified in Table 4, and more preferably encodes a polypeptide selected from the group consisting of GABAB receptor 1d, tyrosine kinase receptor RET and Neurodap-1.  
         [0084]     In another preferred embodiment of this method, the at least one gene is identified in Table 5, and more preferably encodes a polypeptide selected from the group consisting of inhibin alpha-subunit and vesicular transport factor.  
         [0085]     In another preferred embodiment of this method, the at least one gene is identified in Table 6, and more preferably encodes a polypeptide selected from the group consisting of GABAB receptor 1c and phosphatidylinositol 4-kinase.  
         [0086]     In another preferred embodiment of this method, the at least one gene is identified in Table 7 and more preferably encodes a polypeptide selected from the group consisting of somatostain-14 and kainate receptor submit (ka2).  
         [0087]     In another preferred embodiment of this method, the at least one gene is identified in Table 8, and more preferably encodes a polypeptide selected from the group consisting of melanocortin-3 receptor, somatostatin, metabotropic glutamate receptor 3, NCAM polypeptide and synaptic SAPAP-interacting protein.  
         [0088]     In another preferred embodiment of this method, the at least one gene is identified in Table 9, and more preferably encodes calpastatin.  
         [0089]     In another preferred embodiment of this method, the at least one gene is identified in Table 10, and more preferably encodes a polypeptide selected from the group consisting of RAC protein kinase alpha, alpha-2B-adrenergic receptor and SNAP-25A.  
         [0090]     In another preferred embodiment of this method, the at least one gene is identified in Table 11, and more preferably encodes a polypeptide selected from the group consisting of oxytosin/neurophysin, NMDAR2C and GABA-A receptor epsilon.  
         [0091]     In another preferred embodiment of thiis method, the at least one gene is identified in Table 12, and preferably encodes a polypeptide selected from the group consisting of phosphodiesterase I, tyrosine phosphatase and dopamine transporter.  
         [0092]     In yet another preferred embodiment of this method, the at least one gene is identified in Table 13, and preferably encodes synaptotagmin IV homolog.  
         [0093]     In another useful embodiment of this method, the at least one gene is identified in Table 14, and preferably encodes a polypeptide selected from the group consisting of calmodulin, protein kinase rMNK2, phospholipase C-beta1b.  
         [0094]     In another useful embodiment of this method, the at least one gene is identified in Table 15, and preferably encodes a polypeptide selected from the group consisting of phosphatidylinositol 4-kinase and protein-tyrosine-phosphatase.  
         [0095]     A “therapeutically effective amount” of a therapeutic agent refers to a sufficient amount of the therapeutic agent to prevent or inhibit cocaine addiction-related behavior in a subject suffering from cocaine addiction. The determination of a therapeutically effective amount is well within the capability of those skilled in the art. For any therapeutic, the therapeutically effective dose can be estimated in animal models, usually mice, rats, rabbits, dogs or pigs. The animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.  
         [0096]     Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in experimental animals, e.g., ED 50  (the dose therapeutically effective in 50% of the population) and LD 50  (the dose lethal to 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, and it can be expressed as the ratio, LD 50 /ED 50 . Antisense nucleotides, ribozymes, double-stranded RNAs, antagonists and agonists, and other therapeutic agents that exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies is used in formulating a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that include the ED 50  with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, sensitivity of the subject and the route of administration.  
         [0097]     The exact dosage will be determined by the practitioner, in light of factors related to the subject that requires treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, general health of the subject, age, weight and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities and tolerance/response to therapy.  
         [0098]     Normal dosage amounts may vary from 0.1-100,000 mg, up to a total dose of about 1 g, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art will employ different formulations for nucleotides than for antagonists.  
         [0099]     For therapeutic applications, the therapeutic agents are preferably administered as pharmaceutical compositions containing the therapeutic agent in combination with one or more pharmaceutically acceptable carriers. The compositions may be administered alone or in combination with at least one other agent, such as stabilizing compound, which may be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose and water. The compositions may be administered to a subject, or in combination with other agents or drugs.  
         [0100]     In another aspect, the present invention provides screening methods for identifying agents to be tested for the ability to inhibit or prevent cocaine addiction-related behavior. The screening methods are typically designed to find candidate agents that can interact, i.e., bind, to proteins encoded by these differentially expressed genes, and then these agents can be used in assays that ascertain the ability of the candidate agent to modify the activity of the protein. Such binding and activity assays can be performed in cell-free systems, e.g., in a reconstituted protein mixture or a cell membrane preparation, and in cells, particularly recombinant cells expressing the protein encoded by the gene. In particularly useful embodiments of these screening methods, candidate agents are screened in animal models for their ability to reverse, i.e., either increase or decrease, the expression of at least one of the disclosed genes that are upregulated or down regulated by cocaine withdrawal.  
         [0101]     As used herein, the term “candidate agent” refers to any molecule that is capable of interacting, i.e., binding to, and/or increasing or decreasing the activity of, a protein encoded by one of the disclosed genes. The candidate agent can modify the structure of the encoded protein to thereby alter the activity of the protein. The candidate agent also refers to any molecule that is capable of increasing/decreasing the level of mRNA corresponding to or protein encoded by at least one of the disclosed genes. The candidate agent can be natural or synthetic molecules such as proteins or fragments thereof, antibodies, nucleic acid molecules, e.g., antisense nucleotides, ribozymes, double-stranded RNAs, organic and inorganic compounds and the like.  
         [0102]     In one embodiment, cell-free assays for identifying such candidate agents comprise combining in a reaction mixture, i.e, a cell-free system or cell-based system, a candidate agent with a protein encoded by one of the disclosed genes in Tables 1-15 and determining the interaction, i.e., binding, of the candidate agent to the protein or modulation of the activity of the protein. In other embodiments, a fragment of the protein encoded by the disclosed gene can be combined with the candidate agent. Preferred proteins include those encoded by genes identified in Tables 1, 5, 8, 11 and 14. More preferred proteins are those encoded by the preferred listed genes for each of Tables 1, 2, and 4-15, and preferably Table 1 as described above in the methods for inhibiting addiction-related behavior. In some embodiments of this cell-free assay, the gene identified in Table 1 does not encode CB1 cannibinoid receptor.  
         [0103]     In a particularly useful embodiment, the protein encoded by the disclosed gene or the candidate agent is immobilized to an insoluble support to facilitate separation of complexes of the protein/candidate agent from uncomplexed forms of the protein and automation of the assay. The insoluble support may be solid or porous and possess any shape. Examples of suitable solid supports include, but are not limited to, microtitre plates and arrays, micro-centrifuge tubes, test tubes, membranes and beads. Particularly useful methods of binding include, but are not limited to, the use of antibodies, direct binding to ionic supports, and chemical crosslinking. Subsequent to binding of the protein or agent to the support, unbound material is removed by washing.  
         [0104]     In a preferred embodiment, the protein encoded by the gene is bound to the insoluble support, and the candidate agent is then added. Alternatively, the candidate agent is bound to the solid support and the protein encoded by the gene is added.  
         [0105]     Determination of the binding of the candidate agent to the encoded protein can be carried out by standard methods. For example, the candidate agent can be labeled, and binding determined by, e.g, attaching the protein or fragment thereof to the insoluble support, adding the labeled candidate agent, washing off unbound candidate agent, and determining whether any label is bound to the support.  
         [0106]     The term “labeled” means that the candidate agent or protein is either directly or indirectly labeled with a label to provide a detectable signal, e.g., enzymes, antibodies, radioisotopes, fluorescers, chemiluminescers, or specific binding molecule pairs such as biotin and streptavidin. For example, the protein can be biotinylated using biotin NHS(N-hydroxysuccinimide), using well-known techniques and immobilized in the well of streptavidin-coated plates.  
         [0107]     Interaction (binding) between molecules can also be assessed by using real-time BIA (Biomolecular Interaction Analysis, Pharmacia Biosensor AB), which detects surface plasmon resonance, an optical phenomenon. Detection depends on changes in the mass concentration of mass macromolecules at the biospecific interface and does not require labeling of the molecules. In one useful embodiment, a library of candidate agents, such as organic compounds, can be immobilized on a sensor surface, e.g., a wall of a micro-flow cell. A solution containing the protein or functional fragment thereof is then continuously circulated over the sensor surface. An alteration in the resonance angle, as indicated on a signal recording, indicates the occurrence of an interaction. This technique is described in more detail in BIAtechnology Handbook by Pharmacia.  
         [0108]     In another embodiment, the binding of the candidate agent to the protein encoded by the gene can be determined using competitive binding assays wherein a competitor, i.e., a substance known to bind to the encoded protein such as an antibody, ligand, peptide, etc., is combined with the encoded protein, either prior to or subsequent to combining the protein with the candidate agent. For example, the competitor can be added to the protein followed by the candidate agent. Displacement of the competitor indicates that the candidate agent is binding to the encoded protein. In this embodiment, the candidate agent or competitor can be labeled. Accordingly, if a labeled competitor is used, the presence of the label in the wash removed from the insoluble support, indicates displacement by the candidate agent. Alternatively, if the candidate agent is labeled, the presence of the label on the insoluble support indicates displacement of the competitor.  
         [0109]     Cell-free assays can also be used to identify agents which interact with a protein encoded by one of the disclosed genes and modulate the activity of this protein. In one embodiment, the protein encoded by one of the disclosed genes is incubated with a candidate agent, such as an organic compound and the catalytic activity of the protein is determined.  
         [0110]     In another aspect, a cell-based assay is provided for screening candidate agents that bind to a protein encoded by one of the disclosed genes. The method comprises providing a recombinant cell expressing a protein encoded by one of the genes identified in Tables 1-15, contacting the cell with a candidate agent; and determining the binding of the candidate agent to the protein. As used herein, the term “recombinant cell” refers to a cell that has been transfected by one of the disclosed genes, wherein the cell expresses the gene. The recombinant cell is preferably a mammalian cell, an insect cell, a  xenopus  cell or an oocyte. Cells used as controls include cells that are substantially identical to the recombinant cells, but do not express the proteins encoded by the disclosed genes. The binding of the candidate agent to the protein expressed by the cell can be determined by e.g., detecting a signal in the cell, e.g., alterations in second messengers which are sensitive to binding of the candidate agent. Such a recombinant cell further comprises a reporter gene operatively linked to a transcriptional control sequence which is responsive to an intracellular signal, i.e., a second messenger, transduced by interaction of the candidate agent with the protein expressed by the recombinant cell. For example, cyclic AMP accumulation induced by CB1 activation can be measured using a cyclic AMP response element (CRE) reporter assay. Candidate agents that enhance or suppress expression of the reporter interact with either CB1 or its signal transduction system.  
         [0111]     The term “transcriptional control sequence” refers to DNA sequences, such as initiator sequences, enhancer sequences and promoter sequences, which induce, repress or ortherwise control the transcription of protein encoding nucleic acid sequence to which they are operatively linked. Upon induction of the transcriptional control sequence by the second messenger, the reporter gene is expressed thereby providing a quantifiable and detectable signal, e.g., color, fluorescence, luminescence, cell growth, drug resistance, etc., that determines binding of the candidate agent to the protein. Examples of such reporter genes include, but are not limited to, luciferase, alkaline phosphatase, chloramphenicol acetyl transferase and betagalactosidase. In some embodiments, the protein encoded by one of the genes identified in Table 1 is not CB1 cannabinoid receptor. In some embodiments, modulation of binding of the protein encoded by one of the disclosed genes to the candidate agent can be determined in the presence of a target protein or target peptide which is known to bind to the a protein encoded by one of the disclosed genes.  
         [0112]     In yet another embodiment, the effect of a candidate agent on the transcription of one of the genes disclosed in Tables 1-15 is determined by transfection experiments using a reporter gene operatively linked to at least a portion of a transcriptional control sequence of a gene identified in Tables 1-15.  
         [0113]     Assays based on animal models or cells obtained from such animals can also be used to identify agents which modulate the expression of a gene identified in Tables 1-15, that has undergone up- or down-regulation upon cocaine-withdrawal. Accordingly, in one embodiment, a method for identifying an agent to be tested for an ability to prevent or inhibit addiction related-behavior is provided which comprises: 
        a) exposing a cocaine-addicted subject or brain cells of a cocaine-addicted subject to a candidate agent, wherein the cocaine-addicted subject is undergoing withdrawal;     b) determining a level of expression of at least one gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject, wherein the at least one gene is identified in Tables 1-15; and     comparing the level of expression of the gene in both the cocaine-addicted subject or brain cells of the cocaine-addicted subject in the presence of the candidate agent with the level of expression of the gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject in the absence of the candidate agent, wherein a reversal in the level of expression of the gene in the cocaine-addicted subject or brain cells of the cocaine-addicted subject in the presence of the candidate agent relative to the level of expression of the gene in the absence of the candidate agent indicates that the candidate agent is an agent to be tested for the ability to prevent or inhibit addiction related behavior.        
 
         [0117]     In some embodiments of the latter method, if at least one gene is detected the gene does not encode melanocortin 4 receptor.  
         [0118]     In another embodiment, a method for identifying an agent to be tested for an ability to prevent or inhibit cocaine addiction-related behavior is provided which comprises: 
        a) contacting a brain tissue sample from each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     b) detecting a level of expression of at least one gene in both tissue samples, wherein the gene encodes a polypeptide selected from the group consisting of hypertension-regulated vascular factor, myelin-associated basic protein, PB cadherin, calcitonin receptor, melanocortin 4 receptor, ALK-7 kinase and retroposon;     c) subtracting the level of expression of the gene in the sample obtained from the cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the subject having cocaine addiction-related behavior to provide a first value;     d) administering a candidate agent to each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     e) detecting a level of expression of the at least one gene in both tissue samples obtained from the subjects treated with the candidate agent;     f) subtracting the level of expression of the gene in the sample obtained from the treated cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the treated subject having the cocaine addiction-related behavior to provide a second value; and     g) comparing the second value with the first value wherein a decreased second value relative to the first value is indicative of an agent useful in preventing or inhibiting the cocaine addiction-related behavior.        
 
         [0126]     In yet another embodiment, a method for identifying an agent to be tested for an abiity to prevent or inhibit cocaine addiction-related behavior is provided which comprises: 
        a) obtaining a brain tissue sample from each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     b) detecting a level of expression of at least one gene in both tissue samples, wherein the gene encodes a polypeptide selected from the group consisting of GABA-B receptor subunit gb2, cell adhesion-like molecule,  bos taurus -like neuronal axonal protein, similar to mouse chemokine-like factor, FRA-2, similar to human oxygen-regulated protein, similar to mouse mrg1 protein, pentraxin, malic enzyme, olfactomedin-related protein, arc-growth factor enriched in dendrites, protein tyrosine phosphatase, krox, neuritin, microtubule-associated protein 2d and CB1 cannabinoid receptor;     c) subtracting the level of expression of the gene in the sample obtained from the cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the subject having cocaine addiction-related behavior to provide a first value;     d) administering a candidate agent to each of a subject having a cocaine addiction-related behavior and a cocaine addiction-free subject;     e) detecting a level of expression of at least one gene in both tissue samples obtained from the subjects treated with the candidate agent;     f) subtracting the level of expression of the gene in the sample obtained from the treated cocaine addiction-free subject from the level of expression of the gene in the sample obtained from the treated subject having the cocaine addiction-related behavior to provide a second value; and     g) comparing the second value with the first value wherein an increased second value relative to the first value is indicative of an agent useful in preventing or inhibiting the cocaine addiction-related behavior.        
 
         [0134]     The level of expression of at least one of the disclosed genes in the samples obtained from the subject and disease-free subject and brain cells obtained from the subjects can be detected by measuring either the level of mRNA corresponding to the gene or the protein encoded by the gene. RNA can be isolated from the samples by methods well-known to those skilled in the art as described e.g., in Ausubel et al., Current Protocols in Molecular Biology, Vol. 1, pp. 4.1.1-4.2.9 and 4.5.1-4.5.3, John Wiley &amp; Sons, Inc. (1996).  
         [0135]     Methods for detecting the level of expression of mRNA are well-known in the art and include, but are not limited to, Northern blotting, reverse transcription PCR, real time quantitative PCR and other hybridization methods.  
         [0136]     A particularly useful method for detecting the level of mRNA transcripts obtained from a plurality of the disclosed genes involves hybridization of labeled mRNA to an ordered array of oligonucleotides. Such a method allows the level of transcription of a plurality of these genes, i.e., two or more, to be determined simultaneously to generate gene expression profiles or patterns. The gene expression profile derived from the sample obtained from the subject having the cocaine addiction-related behavior treated with agent can be compared with the gene expression profile derived from the sample obtained from the untreated subject having the cocaine addiction-related behavior to determine whether the genes are up- or down-regulated in the sample from the treated subject relative to the genes in the sample obtained from the untreated subject, and thereby determine whether the agent prevents or inhibits cocaine addition-related behavior.  
         [0137]     The oligonucleotides utilized in this hybridization method are bound to a solid support. Examples of solid supports include, but are not limited to, membranes, filters, slides, paper, nylon, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, polymers, polyvinyl chloride dishes, etc. Any solid surface to which the oligonucletides can be bound, either directly or indirectly, either covalently or non-covalently, can be used. A particularly preferred solid substrate is a high-density array or DNA chip (see “Materials and Methods”; and Example 1). These high density arrays contain a particular oligonucleotide probe in a pre-selected location on the array. Each pre-selected location can contain more than one molecule of the particular probe. Because the oligonucleotides are at specified locations on the substrate, the hybridization patterns and intensities (which together result in a unique expression profile or pattern) can be interpreted in terms of expression levels of particular genes.  
         [0138]     The oligonucleotide probes can be labeled with one or more labeling moieties to permit detection of the hybridized probe/target polynucleotide complexes. Label moieties can include compositions that can be detected by spectoscopic, biochemical, photochemical, bioelectronic, immunochemical, electrical optical or chemical means. Examples of labeling moieties include, but are not limited to, radioisotopes, e.g.,  32 P,  33 P,  35 S, chemiluminescent compounds, labeled binding proteins, heavy metal atoms, spectoscopic markers such as fluorescent markers and dyes, linked enzymes, mass spectrometry tags and magnetic labels.  
         [0139]     Oligonucleotide probe arrays for expression monitoring can be prepared and used according to techniques which are well-known to those skilled in the art as described, e.g., in Lockhart et al., Nat. Biotech., Vol. 14, pp. 1675-1680 (1996); McGall et al., Proc. Natl. Acad. Sci. USA, Vol. 93, pp. 13555-13460 (1996); and U.S. Pat. No. 6,040,138.  
         [0140]     Expression of the protein encoded by the gene(s) can be detected by a probe which is detectably labeled, or which can be subsequently labeled. Generally, the probe is an antibody or other ligand which recognizes the expressed protein.  
         [0141]     As used herein, the term “antibody” includes, but is not limited to, polyclonal antibodies, monoclonal antibodies, humanized or chimeric antibodies, and biologically functional antibody fragments which are those fragments sufficient for binding of the antibody fragment to the protein.  
         [0142]     For the production of antibodies to a protein encoded by one of the disclosed genes, various host animals may be immunized by injection with the polypeptide, or a portion thereof. Such host animals may include, but are not limited to, rabbits, mice and rats, to name but a few. Various adjuvants may be used to increase the immunological response, depending on the host species, including but not limited to Freund&#39;s (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances, such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, dinitrophenol and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and  Corynebacterium parvum.    
         [0143]     Polyclonal antibodies are heterogeneous populations of antibody molecules derived from the sera of animals immunized with an antigen, such as target gene product, or an antigenic functional derivative thereof. For the production of polyclonal antibodies, host animals, such as those described above, may be immunized by injection with the encoded protein, or a portion thereof, supplemented with adjuvants as also described above.  
         [0144]     Monoclonal antibodies (mAbs), which are homogeneous populations of antibodies to a particular antigen, may be obtained by any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique of Kohler et al., Nature, Vol. 256, pp. 495-497 (1975) and U.S. Pat. No. 4,376,110, the human B-cell hybridoma technique (see Kosbor et al., Immunology Today, Vol. 4, p. 72 (1983); Cole et al., Proc. Natl. Acad. Sci. USA, Vol. 80, pp. 2026-2030 (1983); and the EBV-hybridoma technique (see Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96 (1985)). Such antibodies may be of any immunoglobulin class including IgG, IgM, IgE, IgA, IgD and any subclass thereof. The hybridoma producing the mAb of this invention may be cultivated in vitro or in vivo. Production of high titers of mAbs in vivo makes this the presently preferred method of production.  
         [0145]     In addition, techniques developed for the production of “chimeric antibodies” (Morrison et al., Proc. Natl. Acad. Sci. USA, Vol. 81, pp. 6851-6855 (1984); Neuberger et al., Nature, Vol. 312, pp. 604-608 (1984); Takeda et al., Nature, Vol. 314, pp. 452-454 (1985)) by splicing the genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used. A chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable or hypervariable region derived from a murine mAb and a human immunoglobulin constant region.  
         [0146]     Alternatively, techniques described for the production of single chain antibodies (see U.S. Pat. No. 4,946,778; Bird, Science, Vol. 242, pp. 423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA, Vol. 85, pp. 5879-5883 (1988); and Ward et al., Nature, Vol. 334, pp. 544-546 (1989)) can be adapted to produce differentially expressed gene single-chain antibodies. Single-chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single-chain polypeptide.  
         [0147]     Most preferably, techniques useful for the production of “humanized antibodies” can be adapted to produce antibodies to the proteins, fragments or derivatives thereof. Such techniques are disclosed in U.S. Pat. Nos. 5,932,448; 5,693,762; 5,693,761; 5,585,089; 5,530,101; 5,569,825; 5,625,126; 5,633,425; 5,789,650; 5,661,016; and 5,770,429.  
         [0148]     Antibody fragments which recognize specific epitopes may be generated by known techniques. For example, such fragments include, but are not limited to, the F(ab′) 2  fragments which can be produced by pepsin digestion of the antibody molecule and the Fab fragments which can be generated by reducing the disulfide bridges of the F(ab′) 2  fragments. Alternatively, Fab expression libraries may be constructed (see Huse et al., Science, Vol. 246, pp. 1275-1281 (1989)) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity.  
         [0149]     The extent to which the known proteins are expressed in the sample is then determined by immunoassay methods which utilize the antibodies described above. Such immunoassay methods include, but are not limited to, dot blotting, Western blotting, competitive and non-competitive protein binding assays, enzyme-linked immunosorbant assays (ELISA), immunohistochemistry, fluorescence-activated cell sorting (FACS) and others commonly used and widely described in scientific and patent literature, and many employed commercially.  
         [0150]     Particularly preferred, for ease of detection, is the sandwich ELISA, of which a number of variations exist, all of which are intended to be encompassed by the present invention. For example, in a typical forward assay, unlabeled antibody is immobilized on a solid substrate and the sample to be tested brought into contact with the bound molecule. After a suitable period of incubation, for a period of time sufficient to allow formation of an antibody-antigen binary complex. At this point, a second antibody, labeled with a reporter molecule capable of inducing a detectable signal, is then added and incubated, allowing time sufficient for the formation of a ternary complex of antibody-antigen-labeled antibody. Any unreacted material is washed away, and the presence of the antigen is determined by observation of a signal, or may be quantitated by comparing with a control sample containing known amounts of antigen. Variations on the forward assay include the simultaneous assay, in which both sample and antibody are added simultaneously to the bound antibody, or a reverse assay in which the labeled antibody and sample to be tested are first combined, incubated and added to the unlabeled surface bound antibody. These techniques are well-known to those skilled in the art, and the possibility of minor variations will be readily apparent. As used herein, “sandwich assay” is intended to encompass all variations on the basic two-site technique. For the immunoassays of the present invention, the only limiting factor is that the labeled antibody be an antibody which is specific for the protein expressed by the gene of interest.  
         [0151]     The most commonly used reporter molecules in this type of assay are either enzymes, fluorophore- or radionuclide-containing molecules. In the case of an enzyme immunoassay an enzyme is conjugated to the second antibody, usually by means of glutaraldehyde or periodate. As will be readily recognized, however, a wide variety of different ligation techniques exist, which are well-known to the skilled artisan. Commonly used enzymes include horseradish peroxidase, glucose oxidase, beta-galactosidase and alkaline phosphatase, among others. The substrates to be used with the specific enzymes are generally chosen for the production, upon hydrolysis by the corresponding enzyme, of a detectable color change. For example, p-nitrophenyl phosphate is suitable for use with alkaline phosphatase conjugates; for peroxidase conjugates, 1,2-phenylenediamine or toluidine are commonly used. It is also possible to employ fluorogenic substrates, which yield a fluorescent product rather than the chromogenic substrates noted above. A solution containing the appropriate substrate is then added to the tertiary complex. The substrate reacts with the enzyme linked to the second antibody, giving a qualitative visual signal, which may be further quantitated, usually spectrophotometrically, to give an evaluation of the amount of protein which is present in the serum sample. Alternately, fluorescent compounds, such as fluorescein and rhodamine, may be chemically coupled to antibodies without altering their binding capacity. When activated by illumination with light of a particular wavelength, the fluorochrome-labeled antibody absorbs the light energy, inducing a state of excitability in the molecule, followed by emission of the light at a characteristic longer wavelength. The emission appears as a characteristic color visually detectable with a light microscope. Immunofluorescence and EIA techniques are both very well-established in the art and are particularly preferred for the present method. However, other reporter molecules, such as radioisotopes, chemiluminescent or bioluminescent molecules may also be employed. It will be readily apparent to the skilled artisan how to vary the procedure to suit the required use.  
         [0152]     The following examples are included to demonstrate preferred embodiments of the invention.  
       EXAMPLES  
       [0000]     Research Design and Methods  
         [0153]     Strategy to Identify Changes in Gene Expression in the NAc Shell and other Brain Regions During Prolonged Abstinence  
         [0154]     Six groups of rats (n=10/group) underwent 3 weeks (15 days) of daily (6-10 hours) cocaine self-administration, followed by short or long periods of forced abstinence prior to sacrifice. Changes in gene expression that coincide with time-dependent increases in cocaine-seeking behavior were identified by comparing changes in 1-week abstinence and 1-week extinction groups, as illustrated in  FIG. 5  below (Groups I and II, respectively). First, a direct comparison between 1-week abstinence and 1-week extinction groups was conducted to identify differences. This allowed detection of genes that correspond to the groups with the greatest differences in drug-seeking behavior. Second, each experimental group (1 week abstinence and extinction) was directly compared to their respective untreated control groups to test whether differences between the groups represent reversals in gene expression between the withdrawal and extinction conditions. Direct comparisons with control groups also allowed detection of genes changed in withdrawal or extinction that might also contribute to drug-seeking behavior though their levels might not necessary be reversed between extinction and withdrawal.  
         [0155]     Surgery, Behavioral Testing and Dissection of Specific Brain Regions  
         [0156]     Both experimental and control groups consisted of individually housed, male Sprague Dawley rats. Experimental animals were surgically implanted with chronic, indwelling intravenous catheter as follows (see Sutton et al., supra). All surgery were performed under aseptic conditions, in a clean area used solely for surgical procedures. Each surgery was done on a separate, clean sheet of Whatman Benchkote paper. Surgical instruments were autoclaved and cleaned (cleaned and soaked in 70% ethanol between successive surgeries). Rats (at least 300 g) and mice (25-30 g) were anesthetized with an i.p. injection of pentobarbital (1.0 mg/kg; rats) and ketamine/xylazine (10 mL/kg; mice), and penicillin procaine intramuscular (i.m.) (60,000 IU/0.2 mL rats, 6,000 units/0.02 mL mice) was given as a prophylactic. The back area of the animals were shaved and cleaned with 70% ethanol, and 2 incisions were made, one on the back (2 cm), and one on the neck (1 cm). The jugular vein was isolated and a sterile Silastic catheter was inserted to the level sinus just outside the right atrium, and mounted in place with surgical mesh. The remaining catheter was pulled from the neck area subcutaneously back incision. Then the catheter exited via 22-gauge stainless steel tubing cemented into place with dental cement and skull screws on a plastic back mount. The incisions were sutured closed with silk surgical thread and the wounds treated with topical antibiotic, and the animal were given an i.m. injection of penicillin G procaine i.m. (60,000 IU/0.2 mL).  
         [0157]     Rats implanted with intravenous (i.v.) catheter recovered from surgery on a warming pad. The rats were not used for experimentation for at least 4 days. During this time, each animal was monitored for distress or infection, and the catheter was flushed daily with 0.2 mL of heparinized saline (20 IU/mL/kg). Because prior exposure to analgesics can alter subsequent behavioral responses to drugs of abuse, rats did not receive post-operative analgesics. Controls remained in their home cages with frequent handling throughout the experiment. Experimental rats were allowed to self-administer cocaine by lever pressing (Fixed-ratio 1: Time-out 10 seconds, 0.5 mg/kg/injection) during their dark cycle 5 days/week for 3 weeks. Each cocaine infusion was delivered over 1.25 seconds concurrent with a cue light, and followed by a 10-second time-out period. The house-light was extinguished during the injection time-out period; together these stimuli constituted the cocaine cue used in reinstatement below. The experimental animals self-administered cocaine in contextually distinct operant chambers located in testing rooms outside the animal colony. During the first week, rats self-administered cocaine for 10 hours/day to hasten acquisition and accustom them to high levels of cocaine exposure. During the second and third weeks, animals self-administered cocaine 6 hours/day. These conditions typically produced self-regulated levels of cocaine intake of 50-60 mg/kg/6-hour test session at the end of self-administration testing, and more precisely mimic daily patterns of cocaine binges in humans.  
         [0158]     Following 3 weeks of cocaine self-administration, animals were divided into experimental groups with equivalent mean levels of cocaine intake, and important factor that determines the propensity for cocaine-seeking during abstinence. Experimental Group II underwent extinction training for 5 days during the first week of abstinence for 6 hours/day, beginning 3 days after their final self-administration test session. Experimental Group IV underwent extinction training for 5 days during their sixth week of abstinence. Responding at both drug-paired and inactive lever were recorded during this time. On the last hour of the final extinction test session, cue-induced reinstatement of cocaine-seeking behavior was tested. During this hour, cues specifically associated with prior cocaine infusions during self-administration (house light off/cue light on) were presented every 2 minutes, and responding at the drug-paired and inactive levers were recorded. Experimental Group I remained in their home cages until sacrifice. Three more experimental groups underwent saline self-administration for 3 weeks and were sacrificed along with Groups I, II and IV. Each group consisted of 6-14 animals to reduce the effects of variability from individuals or dissection procedures on array profiling.  
         [0159]     Animals undergoing extinction training were sacrificed 3 days after their last extinction training session; animals remaining in their home cages were sacrificed at similar times during abstinence. Animal were removed from their home cages and immediately sacrificed by decapitation. Brains were rapidly dissected and chilled slices in ice-cold artificial cerebral spinal fluid for 2 minutes. Tissue punches (12- to 16-gauge) were collected from serial coronal brain slices (1.2-1.5 mm thick) based on the locations depicted in  FIG. 6 . A 14-gauge punch was used to collect NAc core samples, and a 12-gauge punch was used to collect a “half moon” slice of the remaining NAc shell tissue, both yielding about 8-10 mg tissue/punch. Punches were rapidly frozen on dry ice, and stored at −80° C. until shipped to GNF for the GeneChip studies.  
         [0160]     Isolation of Total RNA and Synthesis of cRNA Samples  
         [0161]     Total RNA was isolated from pooled tissue samples using Trizol reagent (1 mL Trizol per 50 mg tissue) (Gibco BRL) and a homogenizer (Polytron, Kinematica) run at maximum speed for 90 seconds. The standard Trizol procedure was used, and RNA after ethanol precipitation was further purified with Rneasy columns (Qiagen). Quality of total RNA was assessed by agarose gel electrophoresis and quantity by spectrophotometer in water and Tris, pH 7.5. Yields were lower than expected and ranged from 4-20 μg. After gel electrophoresis and quantitation, the amount of the limiting sample was 3 μg. Due to the low yield, 250 nanogram aliquots were removed as a preventative measure in case cRNA yields were inadequate and a double amplification of the total RNA was needed. Complementary DNA (cDNA) was synthesized from 3 mg total RNA (corresponding to the amount of the sample with lowest yield) using a T7 promotor/oligo dT primer which allows for subsequent linear amplification of the resulting cDNA (see Van Gelder et al., “Amplified RNA Synthesized From Limited Quantities of Heterogeneous cDNA”, Proc. Natl. Acad. Sci. USA, Vol. 87, pp. 1663-1667 (1990)). This procedure results in cDNA and cRNA populations that accurately and reproducibly represent the total RNA of origin (see Lipshutz et al., “High Density Synthetic Oligonucleotide Arrays”, Nature Gen., Vol. 21, pp. 20-24 (1999); Lockhart et al., “Expression Monitoring by Hybridization to High-Density Oligonucleotide Arrays”, Nature Biotech., Vol. 14, pp. 1675-1680 (1996); and Wodicka et al., “Genome-Wide Expression Monitoring in  Saccharomyces cerevisiae ”, Nature Biotech., Vol. 15, pp. 1359-1367 (1997)). Briefly, 3 μg total RNA was used to make first strand cDNA using the Superscript Choice system (Gibco BRL) and a T7 promotor/oligodT primer (Gibco). Second strand cDNA was made with the Superscript Choice system. All of the resulting cDNA, after phenol:chloroform purification and ammonium acetate precipitation, was used as a template to make biotinylated amplified antisense cRNA using T7 RNA polymerase (Enzo kit, Affymetrix). Twenty micrograms cRNA was fragmented to a target range of 20-100 bases in length using fragmentation buffer (200 mM Tris-acetate, pH 8.1, 500 mM KOAc, 150 mM MgOAc) and heating for 35 minutes at 94° C. This procedure both reduces secondary structure of cRNA and prevents it from hybridizing to adjacent DNA probes on the array (Lockhart et al., supra and Southern et al., “Molecular Interactions on Microarrays”, Nature Gen., Vol. 21, pp. 5-9 (1999)). Quality of cRNA and size distribution of fragmented cRNA was examined by both agarose and polyacrylamide gel electrophoresis. It was determined that fragmentation did not yield the expected size range, and further fragmentation resulted in loss of sample. For this reason, the double amplification protocol was used.  
         [0162]     Amplification and Labeling of Small Amounts of mRNA  
         [0163]     Occasionally, yields of total RNA from small amounts of dissected brain regions is poor in quantity and yet of high quality. Thus, we used double linear amplification procedure as described (see Luo et al., “Gene Expression Profiles of Laser-Captured Adjacent Neuronal Subtypes”, [published erratum appears in  Nat. Med., Vol.  5, No. 3, p. 355 (1999)] Nat. Med., Vol. 5, pp. 117-122 (1999)) and modified for use in our laboratory. First and second stranded cDNA was synthesized as described above using 50 ng starting total RNA, but first, unlabeled cRNA was made using the Megascript kit (Ambion). cRNA was purified with a microcon-50 column (Millipore) and cDNA was again made with random primers and Superscript II (GibcoBRL) at 37° C. for 1 hour, incubated at 37° C. in the presence of RNAse H (GibcoBRL) for 20 minutes. After heat denaturing the enzymes, a T7-oligo dT primer was added to the mixture and second strand cDNA was made with DNA polymerase I and then T4 DNA polymerase (GibcoBRL). cDNA was purified with microcon-50 columns (Millipore) and a second round of cRNA amplification was performed using the Enzo kit (Affymetrix). Unlike amplification by PCR, this method results in a linear amplification of the total RNA (above references). Between 39 and 84 μg of labeled cRNA was made from 50 ng starting total RNA. Twenty μg cRNA was fragmented as described above, fragmention was successful as determined by gel electrophoresis, and 15 μg fragmented cRNA was added to Affymetrix Gene Chip® Rat Genome U34 arrays with 1×MES hybridization buffer using standard protocols outlined in the Gene Chip® Expression Analysis Technical Manual (Affymetrix). Hybridization was for 16 hours at 45° C. The same hybridization samples were then removed from the chips and re-hybridized to identical arrays to make duplicates of each sample.  
         [0164]     Washing, Staining and Scanning Arrays  
         [0165]     Following hybridization of sample to arrays, sample was removed and arrays were washed to remove excess sample. Biotinylated cRNA that is specifically hybridized to the array was stained first with streptavidin phycoerythrin (SAPE, Molecular Probes), then with biotinylated anti-streptavidin antibody, and again with SAPE using standard protocols outlined in the Gene Chip® Expression Analysis Technical Manual (Affymetrix). Following washing, arrays were scanned with a laser scanner (Agilent). After scanning, Gene Chip® software aligns a grid to the image so that individual probe sets can be identified. The quantitative assessment of “present” or “absent” probe sets is based on the number of instances in which the PM signal is significantly larger than the MM signal across the redundant set of probes for each gene. This array design and analysis scheme is essentially a “voting” scheme. Determination of quantitative RNA abundance is made from the average of the pairwise differences (PM minus MM) across the set of probes for each RNA (average difference value). In order to compare average difference values for each RNA between different arrays, intensity values are scaled (normalized) using intensity values taken over the entire array. The Gene Chip® software makes qualitative calls of “Increase” or “Decrease” and quantitative assessments of the absolute size (“fold change”) of any differences. In order to increase confidence in the results, all experiments were performed using duplicate hybridizations. Only differences between duplicates are considered (see below).  
         [0166]     Data Filtering to Find Differentially Expressed Genes (Primary Screen)  
         [0167]     We have developed a Web-based software tool at our institute for gene expression array data filtering. This tool allows us to filter data with user-defined criteria. For example, if one is comparing gene expression changes between arrays A and B, fold changes are first made between A and B. Fold changes are also measured between duplicate arrays A′ and B′. Gene expression changes that are common between the duplicate comparisons are then selected. The criteria for valid differences are as follows: 
        Genes scored as “Increased”/“Moderately Increased” or “Decreased”/“Moderately Decreased” (by the standard Affymetrix algorithm) in both comparisons.     Genes with a minimum 2-fold change in both comparisons, and a minimum absolute change of 50 units in both comparisons.     Genes scored as “present” in the experimental file or “present” or “moderate” in the baseline file of at least one of the two comparisons.        
 
         [0171]     This software tool can rapidly and accurately manage thousands of potentially regulated genes with a variety of filter settings. The stringency of the filter can be varied depending on the number of potentially regulated genes found. This same data filtering tool can also be used to examine the consistency of the duplicate arrays by finding the number of genes that are significantly “different” between duplicates.  
         [0172]     A different data filtering approach was used to find differentially expressed genes in the NAc core, CeA, mPFC and VTA. The reasons for the change in the approach are that the new methods are easily adaptable to our gene expression database and they do not rely on “Increase, Decrease, Absence or Presence” calls generated by the Affymetrix algorithm. The Web-based tool used for finding gene expression changes in the NAc shell is less practical to use.  
         [0173]     Two different filters were used to generate data for the NAc core, CeA, mPFC and VTA. The sum of the findings from both filters were used to generate the final gene lists, with redundant entries collapsed to generate one entry per probe set. The first filter used was a one-way ANOVA. Values less than a value of 20 were first forced to a value of 20, then ANOVA was performed.  
         [0174]     Probe sets were retained in the gene lists only after they met the following criteria: 
        1. P-value less than 0.01.     2. Fold change difference between statistical groups at least 1.7.     3. Maximum intensity (average difference value) across the group of at least a value of 200.        
 
         [0178]     The second filter used to generate data for the NAc core, CeA, mPFC and VTA avoided the potential problems of using ANOVA for small sample sizes. First, all values less than a value of 200 were forced to a value of 200. Then, mean values of the groups, standard deviations within the groups, and fold change differences between the groups were calculated and probe sets were retained only if they met the following criteria: 
        1. Fold change difference between groups at least 1.7.     2. The standard deviation of the group divided by the mean of the same group must have been a value of 0.25 or less for both groups.        
 
         [0181]     Tissue Dissection/Western Blot Procedures  
         [0182]     Rats were removed from their homecages and immediately decapitated in a separate room; the brains were rapidly dissected and chilled in ice-cold physiological buffer (5 mM KCl, 126 mM NaCl, 1.25 mM NaH 2 PO 4 , 10 mM D-glucose, 25 mM NaHCO 3 , 2 mM CaCl 2 , 2 mM MgSO 4 , pH 7.4). NAc core samples were obtained with a 14-gauge punch from chilled coronal brain slices (0.7-2.2 mM anterior to bregma; Paxinos et al. (1998)), and immediately frozen and stored at −80° C. Half moon-shaped NAc shell samples were obtained with a 12-gauge punch of the remaining ventral-medial shell tissue.  
         [0183]     Tissue samples were homogenized by sonication in 350 μL (NAc) of 1% SDS. Protein concentrations were determined (Lowry et al. (1951)), and 10 μg protein/sample was subjected to SDS-polyacrylamide gel electrophoresis (7.5-10% acrylamide/0.12% bisacrylamide), followed by electrophoretic transfer to nitrocellulose (Bio-Rad, Hercules, Calif.). Proteins were immunolabeled overnight at 4× in blocking buffer consisting of 5% non-fat dried milk powder in PBST (10 mM sodium phosphate, pH 7.4, 0.9% NaCl, 0.1% Tween-20). Following incubation with the primary antibody, blots were washed with blocking buffer, and incubated for 2 hours at 20° C. with horseradish peroxidase-conjugated goat anti-rabbit IgG (1:2000; Chemicon, Temecula, Calif.) in PBST. The blots were washed again in PBST, and immunoreactivity visualized using enhanced chemiluminescense for peroxidase labeling (New England Nuclear, Boston, Mass.). Protein immunoreactivity was quantified by densitometric analysis using NIH Image 1.57 (National Institute of Health, Bethesda, Md.). TH immunoreactivity was linear over a 4-fold range of tissue concentrations under these conditions.  
         [0184]     Data Analysis  
         [0185]     Each gel contained 7-11 control samples alternating with samples from experimental animals. To normalize data from different gels, protein immunoreactivity for each control and experimental sample was expressed as a percentage of the mean control value for that particular gel. For statistical analysis, age- and batch-matched control values were pooled into a single group, and compared with 2 cocaine-trained groups with 1-way ANOVA. Post-hoc comparisons were made among control and cocaine-trained groups with Newman Keuls tests.  
         [0186]     Analysis of data from nucleus accumbens core, central nucleus of the amygdala, medial prefrontal cortex, and ventral tegmental area indicated that the 1 week withdrawal control and 1 week extinction control groups were not equivalent. Therefore pooling all of the control values into a single control group was not valid for these comparisons. Instead, extinction and withdrawal groups were compared directly or to their respective controls.  
       Example 1  
     Identification of Extinction/Withdrawal Differences in Gene Expression in the Nac Shell and Other Brain Regions During Prolonged Abstinence Using Gene Expression Profiling  
       [0187]     The advent of oligonucleotide arrays increases the feasibility of forward genetic approaches to identify gene regulation in studies of complex behaviors. This technology replaces more cumbersome methods of subtraction hybridization and differential display with the advantage of profiling thousands of genes simultaneously.  FIG. 4  illustrates 2 candidate genes identified in our preliminary studies from contralateral NAc shell tissue samples taken from animals used in the extinction studies described above. These genes were selected by comparing 1-week extinction training and 1-week withdrawal groups according to stringent criteria described in the Research Design and Methods section. The top panel illustrates a 3.7-fold difference in expression of a retroviral derived gene retroposon (see Table 1). This gene is over-expressed in withdrawal from cocaine self-administration (88%), but down-regulated (49%) in animals that experienced extinction training when compared to untreated age- and batch-matched controls. In contrast, expression of the CB1 cannabinoid receptor gene is reduced (53%) in withdrawal, but normalized to near control levels following extinction training. Tables 1-15 contain all of the genes selected by both primary and secondary screening procedures for this comparison (see “Methods”). This procedure employs control/control comparisons to eliminate false positives, in addition to the gene filtering software-based selection procedure. As shown in Table 1, there are several genes for structural proteins (i.e., PB cadherin, microtubule-associated protein) suggesting neuroplasticity in neuronal contacts (dendritic spines and arborization). There are also 4 gene candidates (highlighted in bold) that already are implicated in drug reward and addiction. For example, GABA B receptor agonists have been proposed as a possible pharmacotherapy for cocaine addiction, and CB1 cannabinoid receptors mediate central effects of cannabis, and can modulate dopaminergic responses in striatum. Similarly, FRA2, is a Fos-Related Antigen like ΔFosB, which has been implicated in sensitivity to cocaine (see Kelz et al., “Expression of the Transcription Factor ΔFosB in the Brain Controls Sensitivity to Cocaine”, Nature, Vol. 401, pp. 272-276 (1999). The melanocortin receptor MC4 has recently been shown to be up-regulated during withdrawal from repeated cocaine treatments, and intra-NAc infusions of an MC4 antagonist reverse the rewarding effects of cocaine to produce a cocaine aversion instead in a place preference paradigm (see Taylor et al., “Role of Melanocortin in Drug Reward”, submitted).  
                                     TABLE 1                           Effects of Extinction Training on Gene Expression in the NAc Shell       Following 1 Week Withdrawal from Cocaine Self-Administration                        Extinction   Genbank           1 Week   1 Week   vs.   Accession       Gene Name   Withdrawal*   Extinction*   Withdrawal   No.                 GABA-B receptor subunit gb2     ↓ 30%   ↑ 48%   2.12-fold Δ   AJ011318.1       Hypertension-regulated vascular factor   ↑ from 0   0   Normalized   AF055714       Myelin-associated basic protein   ↑ 140%   ↓ 7%   2.59-fold Δ   X87900.1       PB cadherin   ↑ 17%   ↓ 56%   2.17-fold Δ   D83349.1       Calcitonin receptor   ↑ 33%   ↓ 80%   6.58-fold Δ   L13041.1       Cell adhesion-like molecule   ↓ 88%   ↑ 6%   8.92-fold Δ   M88709.1       Bos taurus-like neuronal axonal protein   ↓ 36%   ↑ 34%   2.08-fold Δ   U92535.1       Similar to mouse chemokine-like factor   ↓ 47%   ↑ 65%   2.08-fold Δ   AF144754.1         FRA-2     ↓ 66%   ↑ 41%   3.21-fold Δ   X98051.1       Similar to human oxygen regulated   ↓ 37%   ↑ 46%   2.32-fold Δ   AI009098       protein       Similar to mouse mrg1 protein   ↓ 48%   ↑ 87%   3.62-fold Δ   AI014091       Pentraxin   ↓ 70%   ↑ 63%   5.41-fold Δ   U18772       Malic enzyme   ↓ 33%   ↑ 61%   2.39-fold Δ   M26594.1       Olfactomedin related protein   ↓ 38%   ↑ 48%   2.40-fold Δ   U03414       Arc - growth factor enriched in   ↓ 45%   ↑ 21%   2.18-fold Δ   U19866.1       dendrites       Protein tyrosine phosphatase   ↓ 55%   ↑ 13%   2.49-fold Δ   U28938         Melanocortin 4 receptor     ↑ 272%   ↓ 35%   4.21-fold Δ   U67863.1       ALK-7 kinase   ↑ 44%   ↓ 44%   2.57-fold Δ   U69702.1       Krox   ↓ 47%   ↑ 15%   2.19-fold Δ   U75397       Neuritin   ↓ 87%   ↑ 28%   10.1-fold Δ   U88958.1       Microtubule-associated protein 2d   ↓ 17%   ↑ 67%   2.02-fold Δ   X74211.1         CB1 cannabinoid receptor     ↓ 53%   ↑ 19%   2.52-fold Δ   X55812.1       Retroposon   ↑ 88%   ↓ 49%   3.70-fold Δ   U83119.1                 *Expressed as % Δ from mean control value for both groups (n = 5-8 pooled samples/group). Genes selected according to procedure described in Research Design and Methods. Gene names in bold indicate gene products in the NAc implicated in drug reward or addiction.          # Only changes in known genes are shown. Genes are selected based on criteria (see Methods) where both duplicate comparisons between extinction and withdrawal groups        # exceed 2-fold and are directionally similar. Base on this primary selection procedure, a secondary selection procedure eliminates genes when average duplicate values from both control groups vary more than 20% from the overall mean of the control groups. For genes expressed in        # low levels (&lt;100 densitometric units), all control values must lie within 25 units of the overall mean. Average difference values for all groups and their respective control groups are shown in the Appendix tables.           
 
         [0188]     Thus, this latter neuroadaptation represents one difference replicated by alternative means (in situ). Several other genes regulated by withdrawal but not modified by extinction, and by extinction training alone are shown in Tables 2-16 below. These results demonstrate oligonucleotide detection of extinction/withdrawal differences.  
                                                   TABLE 2                           Average Difference Values for 1-Week Extinction Versus 1-Week Extinction Controls                    1-Week                   Extinction   1-Week       Probe Set   Gene Name   Control   Extinction                    AF050659UTR#1_at   Activity and neurotransmitter-induced early 7 mRNA   269   114       AF050659UTR#1_at       347   132       AJ000485_at   CLIP-115 protein   95   168       AJ000485_at       40   153       AJ006971_g_at   DAP-like kinase   184   545       AJ006971_g_at       209   641       D83348_at   Long-type PB cadherin   113   285       D83348_at       135   298       K02248cds_s_at   Somatostatin-14 gene   69   365       K02248cds_s_at       132   460       M13100cds#3_f_at   Long interspersed repetitive DNA sequence   730   348       M13100cds#3_f_at       938   474       M16410_at   Neurokinin B precursor   117   262       M16410_at       110   241       M32062_at   Fcgamma receptor   −19   96       M32062_at       20   75       M55015cds_s_at   Nucleolin gene   49   154       M55015cds_s_at       66   147       M89646_g_at   Ribosomal protein S24   665   1466       M89646_g_at       765   1370       rc_AA799406_at   Genes for 18S, 5.8S and 28S ribosomal rRNAs   244   683       rc_AA799406_at       −42   577       rc_M800039_s_at   Unknown   346   667       rc_M800039_s_at       264   667       rc_AA866419_at   Unknown   59   150       rc_AA866419_at       −26   109       rc_AA875268_at   Similar to  B. taurus  PSST subunit   683   1332       rc_AA875268_at   of NADH: ubiquinone oxidoreduc   655   1361       rc_AA891727_g_at   Unknown   250   542       rc_AA891727_g_at       285   576       rc_AA891796_at   1-cys peroxiredoxin;   412   889       rc_AA891796_at   thiol-specific antioxidant   557   1180           protein       rc_AA892041_at     Homo sapiens  over-expressed breast tumor   768   1481       rc_AA892041_at   protein mRNA   788   1482       rc_AA892123_at   Ribosomal protein L36   280   708       rc_AA892123_at       378   761       rc_AA892864_at   Unknown   54   264       rc_AA892864_at       −2   259       rc_AA924772_at   Growth inhibitory factor-metallothionein homolog   1533   2979       rc_AA924772_at       1577   3108       rc_AI010581_at   11 Kd diazepam binding inhibitor   249   569       rc_AI010581_at       246   570       rc_AI014135_g_at   CDK103   822   340       rc_AI014135_g_at       916   317       rc_AI171844_at   F1-aTPase epsilon subunit   563   1232       rc_AI171844_at       564   1345       rc_AI176460_s_at   32S pre-rRNA 5′ terminal part with 28S rRNA sequence   1640   3545       rc_AI176460_s_at       1728   3573       rc_AI227887_at   Similar to  Mus musculus  CDC42 mRNA   304   7       rc_AI227887_at       334   119       rc_AI639367_at   Unknown   574   63       rc_AI639367_at       605   81       rc_AI639521_at   Unknown   141   2       rc_AI639521_at       109   21       U75392_s_at   B-cell receptor associated protein 37   191   516       U75392_s_at       190   488       X02002_at   Thy-1 gene for cell surface glycoprotein   197   482       X02002_at       251   498       X05472cds#1_s_at   2.4 Kb repeat DNA right terminal region   428   169       X05472cds#1_s_at       311   123       X14671cds_s_at   Liver mRNA for ribosomal protein L26   871   1811       X14671cds_s_at       1050   1949       X53581cds#5_f_at   Long interspersed repetitive DNA sequence   460   125       X53581cds#5_f_at       425   109       X55153mRNA_s_at   RP2 gene for ribosomal protein P2   723   1603       X55153mRNA_s_at       596   1639       X56325mRNA_s_at   Alpha-1 globin gene   1886   3834       X56325mRNA_s_at       1848   4115       X61295cds_s_at   L1 retroposon mRNA   1299   635       X61295cds_s_at       1080   529       X62952_at   Vimentin   −60   117       X62952_at       26   117       X63594cds_g_at   RL/IF-1   −32   121       X63594cds_g_at       48   194       X68283_at   Ribosomal protein L29   703   1462       X68283_at       524   1271       Y13714_at   Osteonectin   174   531       Y13714_at       187   505                  
 
         [0189]     Genes that passed the filtering criteria outlined above for the nucleus accumbens shell are listed. Average difference values (from GeneChip version 3.2) are listed for each gene from each duplicate chip from both the 1 week extinction and 1 week extinction control groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                   TABLE 3                           Average Difference Values for 1-Week Extinction, 1-Week       Withdrawal and Their Corresponding Control Groups                            1-week   1-week               1-week   1-week   extinction   withdrawal       Probe set no.   Gene name   extinction   withdrawal   control   control                    AF055714UTR#1_at   Hypertension-regulated   −14   63   −22   −22       AF055714UTR#1_at   vascular factor   −14   55   −23   −3       AF058795_at   GABA-B receptor subunit gb2   621   309   432   468       AF058795_at       695   311   456   421       D28111_at   Myelin-associated basic   995   2551   879   1037       D28111_at   protein   835   2186   783   1243       D83349_at   PB cadherin   1709   3903   2960   3704       D83349_at       1798   3703   2580   3759       L13040_s_at   Calcitonin receptor   11   149   124   83       L13040_s_at       32   134   129   91       M13100cds#1_at   Long repetitive sequence   895   2774   237   1335       M13100cds#1_at       910   3278   414   1220       M13100cds#1_g_at   Long repetitive sequence   117   297   1564   270       M13100cds#1_g_at       84   293   1181   375       M13100cds#2_s_at   Long repetitive sequence   177   892   195   408       M13100cds#2_s_at       181   781   174   355       M13100cds#3_f_at   Long repetitive sequence   348   926   547   1001       M13100cds#3_f_at       474   1206   483   974       M13100cds#4_f_at   Long repetitive sequence   153   547   730   229       M13100cds#4_f_at       89   420   938   192       M13100cds#5_s_at   Long repetitive sequence   212   802   249   390       M13100cds#5_s_at       157   765   175   342       M13100cds#6_f_at   Long repetitive sequence   273   916   425   766       M13100cds#6_f_at       185   626   384   771       M13101cds_f_at   Unknown   57   371   746   307       M13101cds_f_at       153   588   612   439       M88709_at   Cell adhesion-like molecule   341   65   262   264       M88709_at       230   −1   212   339       rc_AA799423_at   Unknown   79   292   172   168       rc_AA799423_at       71   201   193   244       rc_AA799448_g_at   Unknown   470   90   392   385       rc_AA799448_g_at       462   218   489   347       rc_AA799594_at   Unknown   1974   3970   1692   1572       rc_AA799594_at       1497   3251   2238   2138       rc_AA859536_at   Similar to  Bos taurus  neuronal   3524   1672   2666   2273       rc_AA859536_at   axonal membrane protein   3517   1707   2885   2675       rc_AA874803_g_at   Similar to mouse chemokine-   1515   488   896   802       rc_AA874803_g_at   like factor   1485   479   1023   907       rc_AA875001_at   Unknown   255   −72   213   221       rc_AA875001_at       270   24   224   278       rc_AA875032_at   FRA-2   285   102   193   222       rc_AA875032_at       344   94   263   214       rc_AI009098_at   Highly similar to human   612   292   491   390       rc_AI009098_at   oxygen-regulated protein   537   204   344   346       rc_AI014091_at   Highly similar to mouse mrg1   231   84   36   195       rc_AI014091_at   protein (a cytokine-inducible   269   54   138   165           transcr.       rc_AI014135_g_at   CDK103   340   −20   822   332       rc_AI014135_g_at       317   27   916   368       rc_AI072943_at   Pentraxin   167   48   68   51       rc_AI072943_at       55   −7   51   103       rc_AI073204_at   14-33 protein epsilon   1793   561   1398   440       rc_AI073204_at       1535   587   1340   448       rc_AI171506_at   Malic enzyme   95   28   82   79       rc_AI171506_at       118   61   54   50       rc_AI176710_at   Nuclear orphan receptor   358   62   144   272       rc_AI176710_at       305   54   163   252       rc_AI231445_at   Lysosomal glycoprotein   −80   17   2   17       rc_AI231445_at       −12   39   31   4       rc_AI233362_at   Unknown   919   2280   1405   1088       rc_AI233362_at       1045   2321   1359   1073       rc_AI639088_s_at   Unknown   116   377   353   290       rc_AI639088_s_at       92   350   267   251       rc_AI639118_at   Unknown   143   70   98   119       rc_AI639118_at       130   43   94   128       rc_AI639226_at   Unknown   28   91   65   81       rc_AI639226_at       17   73   90   80       rc_AI639367_at   Unknown   63   530   574   553       rc_AI639367_at       81   453   605   405       rc_AI639484_at   Unknown   1520   509   1243   1265       rc_AI639484_at       1539   612   1194   1385       rc_AI639521_at   Alpha beta crystalline gene   2   99   141   103       rc_AI639521_at       21   84   109   141       rc_H31118_at   Unknown   1247   430   1152   810       rc_H31118_at       1227   492   1176   842       U03414_s_at   Olfactomedin-related protein   1183   534   797   785       U03414_s_at       1194   458   907   731       U03416_at   Olfactomedin-related protein   1184   471   803   737       U03416_at       1186   508   846   859       U19866_at   Arc - a growth factor enriched   815   403   683   594       U19866_at   in dendrites   627   257   549   557       U28938_at   Protein tyrosine phosphatase   461   184   320   416       U28938_at       440   178   426   435       U67863_at   Melanocortin 4 receptor   14   125   21   38       U67863_at       39   98   56   49       U69702_at   ALK-7 kinase   67   188   140   128       U69702_at       80   190   125   132       U75397UTR#1_s_at   Krox   1077   461   964   983       U75397UTR#1_s_at       1010   494   887   793       U83119_f_at   Repetitive DNA sequence   68   393   314   730       U83119_f_at       38   426   379   484       U88958_at   Neuritin   260   40   244   216       U88958_at       257   11   158   192       U95920_at   Precentriolar material   107   233   157   161       U95920_at       102   200   −32   129       X01118_at   Atrial natriuretic polypeptie   109   −15   −34   40       X01118_at       124   −17   10   12       X05472cds#1_s_at   Repeat DNA   169   624   428   422       X05472cds#1_s_at       123   633   311   317       X05472cds#2_at   Repeat DNA   660   1396   931   630       X05472cds#2_at       630   1412   807   627       X05472cds#3_f_at   Repeat DNA   133   968   213   188       X05472cds#3_f_at       100   878   210   195       X07686cds_s_at   Repeat DNA   58   291   121   135       X07686cds_s_at       28   275   112   112       X17682_s_at   Microtubule-associated   649   319   352   414       X17682_s_at   protein   596   298   335   388       X53455cds_s_at   Microtubule-associated   225   33   126   217       X53455cds_s_at   protein   299   76   53   161       X53581cds#5_f_at   Repeat DNA   125   366   460   411       X53581cds#5_f_at       109   471   425   768       X55812complete_seq_at   CB1 Cannabinoid receptor   294   99   208   251       X55812complete_seq_at       268   124   247   240       X61295cds_s_at   Retroposon   635   2177   1299   1181       X61295cds_s_at       529   2128   1080   1022                  
 
         [0190]     Genes that passed the filtering criteria outlined above for the nucleus accumbens shell are listed. Average difference values (from GeneChip version 3.2) are listed for each gene from each duplicate chip from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 4                           CeA 1-Week Extinction to Control                                1-week                                   1-week   1-week   with-               1-Week   with-   extinction   drawal   Mean   Mean       Fold       Probe set no.   Description   extinction   drawal   control   control   control   extinction   Ratio   change                    AB016161cds_i_at   AB016161cds  Rattus     352   460   50   234   406   142   0.349754   −2.9             norvegicus  mRNA for           GABAB receptor 1d,           complete cds       AF010466_s_at   AF010466  Rattus     13   −24   298   411   −5.5   354.5   −64.4545   at least             norvegicus  interferon                               2-fold           gamma (IFN-gamma)           mRNA, complete cds       AF031430_at   AF031430  Rattus     227   229   103   119   228   111   0.486842   −2.1             norvegicus  syntaxin 7           mRNA, complete cds       AF042830_at   AF042830  Rattus     433   361   253   206   397   229.5   0.578086   −1.7             norvegicus  proto-oncogene           tyrosine kinase receptor           Ret (c-ret) mRNA, partial           cds       AF102552_s_at   AF102552  Rattus     416   499   216   238   457.5   227   0.496175   −2.0             norvegicus  270 kDa           ankyrin G isoform mRNA,           partial cds       D13962_g_at   D13962 RATGLUT3 Rat   358   341   177   149   349.5   163   0.466381   −2.1           mRNA for neuron glucose           transporter       D17711cds_s_at   D17711cds RATCSBP Rat   301   296   159   149   298.5   154   0.515913   −1.9           mRNA for dC-stretch           binding protein (CSBP),           complete cds       D21800_g_at   D21800 RATPSRC10 Rat   110   114   269   252   112   260.5   2.325893   2.3           mRNA for proteasome           subunit RC10-II, complete           cds       D26154UTR#1_at   D26154UTR#1 RATRB109   532   427   286   220   479.5   253   0.527633   −1.9           Rat mRNA for RB109           (brain specific protein),           complete cds       D26500_at   D26500 RATDLP9A Rat   277   271   137   161   274   149   0.543796   −1.8           mRNA for dynein-like           protein 9A, partial cds       D82071_at   D82071  Rattus norvegicus     207   196   94   81   201.5   87.5   0.434243   −2.3           mRNA for hematopoietic           prostaglandin D synthase,           complete cds/cds = 192,791/           gb = D82071/gi = 2558504/           ug = Rn.10837/len = 1004       E13644cds_s_at   E13644cds cDNA   313   292   151   165   302.5   158   0.522314   −1.9           encoding Neurodap-1           which is located at the           post-synaptic membrane           thickening regions of           neurons and contains           RING-H2 finger motif       J00771_at   J00771 RATPRNASE Rat   173   139   430   353   156   391.5   2.509615   2.5           pancreatic ribonuclease           mRNA       L07398_at   L07398 RATIGVCL  Rattus     670   687   290   280   678.5   285   0.420044   −2.4             norvegicus  (hybridoma           56R-3) immunoglobulin           rearranged gamma-chain           mRNA variable (V) region,           partial cds       M12112mRNA#3_s_at   M12112mRNA#3   347   244   502   657   295.5   579.5   1.961083   2.0           RATANGA2 Rat           angiotensinogen mRNA,           3′ flank       M34331_at   M34331 Rat 60S ribosomal   733   704   1155   1508   718.5   1331.5   1.853166   1.9           subunit protein L35 mRNA,           complete cds/cds = 47,418/           gb = M34331/gi = 206729/           ug = Rn.3458/len = 451       rc_AI639304_at   Rat mixed-tissue library   542   524   301   325   533   313   0.587242   −1.7             Rattus norvegicus  cDNA           clone rx00157 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AA799489_g_at   rc_AA799489 EST188986   108   −84   373   483   12   428   35.66667   35.7             Rattus norvegicus  cDNA,           3′ end/clone = RHEAB66/           clone_end = 3′/           gb = AA799489/gi = 2862444/           ug = Rn.6193/len = 646       rc_AA799498_at   rc_AA799498 EST188995   375   495   44   47   435   45.5   0.104598   −9.6             Rattus norvegicus  cDNA,           3′ end/clone = RHEAB76/           clone_end = 3′/           gb = AA799498/gi = 2862453/           ug = Rn.3835/len = 683       rc_AA800549_at   rc_AA800549 EST190046   275   316   461   655   295.5   558   1.888325   1.9             Rattus norvegicus  cDNA,           3′ end/clone = RLUAB29/           clone_end = 3′/           gb = AA800549/gi = 2863504/           ug = Rn.22957/len = 491       rc_AA800882_g_at   rc_AA800882 EST190379   204   166   436   403   185   419.5   2.267568   2.3             Rattus norvegicus  cDNA,           3′ end/clone = RLUAM60/           clone_end = 3′/           gb = AA800882/gi = 2863837/           ug = Rn.24136/len = 379       rc_AA818114_at   rc_AA818114 UI-R-A0-am-   210   227   107   101   218.5   104   0.475973   −2.1           g-03-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-A0-am-g-03-           0-UI/clone_end = 3′/           gb = AA818114/gi = 2887994/           ug = Rn.7181/len = 556       rc_AA851403_at   rc_AA851403 EST194171   474   453   296   209   463.5   252.5   0.544768   −1.8             Rattus norvegicus  cDNA,           3′ end/clone = RPLAG17/           clone_end = 3′/           gb = AA851403/gi = 2938943/           ug = Rn.3383/len = 393       rc_AA859643_at   rc_AA859643 UI-R-E0-bs-   597   468   243   137   532.5   190   0.356808   −2.8           a-08-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bs-a-08-0-           UI/clone_end = 3′/           gb = AA859643/gi = 2949163/           ug = Rn.32/len = 482       rc_AA875659_s_at   rc_AA875659 UI-R-E0-ct-   157   285   390   485   221   437.5   1.979638   2.0           h-07-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ct-h-07-0-           UI/clone_end = 3′/           gb = AA875659/gi = 2980607/           ug = Rn.10966/len = 424       rc_AA891222_at   rc_AA891222 EST195025   380   322   150   98   351   124   0.353276   −2.8             Rattus norvegicus  cDNA,           3′ end/clone = RHEAQ71/           clone_end = 3′/           gb = AA891222/gi = 3018101/           ug = Rn.1014/len = 568       rc_AA891940_at   rc_AA891940 EST195743   52   212   385   426   132   405.5   3.07197   3.1             Rattus norvegicus  cDNA,           3′ end/clone = RKIAI82/           clone_end = 3′/           gb = AA891940/gi = 3018819/           ug = Rn.3508/len = 523       rc_AA894292_at   rc_AA894292 EST198095   441   319   215   222   380   218.5   0.575   −1.7             Rattus norvegicus  cDNA,           3′ end/clone = RSPAW06/           clone_end = 3′/           gb = AA894292/gi = 3021171/           ug = Rn.19450/len = 599       rc_AA924772_at   rc_AA924772 UI-R-A1-eb-   98   266   499   691   182   595   3.269231   3.3           f-02-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-A1-eb-f-02-0-           UI/clone_end = 3′/           gb = AA924772/gi = 3071908/           ug = Rn.11325/len = 372       rc_AI070108_at   rc_AI070108 UI-R-Y0-Iu-a-   377   336   164   121   356.5   142.5   0.399719   −2.5           09-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-Y0-Iu-a-09-0-           UI/clone_end = 3′/           gb = AI070108/ug = Rn.16863/           len = 529       rc_AI137421_at   rc_AI137421 UI-R-C2p-ok-   163   193   442   479   178   460.5   2.587079   2.6           c-12-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C2p-ok-c-12-0-           UI/clone_end = 3′/           gb = AI137421/ug = Rn.1485/           len = 556       U04934_s_at   U04934 RNU04934  Rattus     435   421   147   284   428   215.5   0.503505   −2.0             norvegicus  Sprague-           Dawley (CD-1) clone Kc1           Na-Ca exchanger mRNA,           partial cds       U75899mRNA_g_at   U75899mRNA RNU75899   791   664   378   462   727.5   420   0.57732   −1.7             Rattus norvegicus  HSPB2           gene, complete cds       X58830_at   X58830 Rat vgr mRNA/   503   484   275   277   493.5   276   0.559271   −1.8           cds = 0.623/gb = X58830/           gi = 57475/ug = Rn.10436/           len = 1241       Z50052_at   Z50052  R. norvegicus     214   232   71   69   223   70   0.313901   −3.2           mRNA for C4BP beta chain           protein/cds = 265.1041/           gb = Z50052/gi = 899381/           ug = Rn.11151/len = 1091                  
 
         [0191]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and its corresponding control group in the CeA. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 5                           CeA 1-Week Extinction to Withdrawal                                1-week                                   1-week   1-week   with-               1-week   with-   extinction   drawal   Mean   Mean       Fold       Probe set no.   Description   extinction   drawal   control   control   control   extinction   Ratio   change                    AB016161cds_i_at   AB016161cds  Rattus     352   460   50   234   406   142   0.349754   −2.9             norvegicus  mRNA for           GABAB receptor 1d,           complete cds       AB000517_s_at   AB000517  Rattus  sp.   300   387   162   146   343.5   154   0.448326   −2.2           mRNA for CDP-           diacylglycerol synthase,           complete cds       AF015304_at   AF015304  Rattus     434   419   201   200   426.5   200.5   0.470106   −2.1             norvegicus  equilbrative           nitrobenzylthioinosine-           sensitive nucleoside           transporter mRNA,           complete cds/cds = 4.1377/           gb = AF015304/gi = 2656136/           ug = Rn.5814/len = 1766       AF041373_s_at   AF041373  Rattus     468   405   190   8   436.5   99   0.226804   −4.4             norvegicus  clathrin           assembly protein short form           (CALM) mRNA, complete           cds/cds = 25.1818/           gb = AF041373/gi = 2792499/           ug = Rn.10888/len = 1921       AF064856_at   AF064856  Rattus  sp.   332   244   514   540   288   527   1.829861   1.8           7acomp protein mRNA,           complete cds       E00775cds_s_at   E00775cds cDNA encoding   223   261   −84   −133   242   −108.5   −0.44835   2.2           rat cardionatrin precursor       J00771_at   J00771 RATPRNASE Rat   −50   −13   430   353   −31.5   391.5   −12.4286   at least           pancreatic ribonuclease                               2 fold           mRNA       J05167_at   J05167 Rat band 3 Cl-/   512   412   164   159   462   161.5   0.349567   −2.9           HCO 3  exchanger (B3RP3)           mRNA, complete cds/           cds = 34.3717/gb = J05167/           gi = 203088/ug = Rn.9859/           len = 3877       K00996mRNA_s_at   K00996mRNA RATCYP45E   200   236   386   368   218   377   1.729358   1.7           Rat cytochrome p-450e           (phenobarbital-induced)           mRNA, 3′ end       L07380_g_at   L07380 RATGHRFRG   375   435   236   227   405   231.5   0.571605   −1.7             Rattus rattus  (clone pGR2)           growth hormone-releasing           factor receptor mRNA           sequence       L07398_at   L07398 RATIGVCL  Rattus     723   611   290   280   667   285   0.427286   −2.3             norvegicus  (hybridoma           56R-3) immunoglobulin           rearranged gamma-chain           mRNA variable (V) region,           partial cds       M10140_at   M10140 Rat skeletal muscle   43   81   345   410   62   377.5   6.08871   6.1           creatine kinase composite           mRNA, complete cds/           cds = 69.1214/gb = M10140/           gi = 203477/ug = Rn.10756/           len = 1410       M32754cds_s_at   M32754cds RATINHBAB1   297   256   578   655   276.5   616.5   2.229656   2.2           Rat inhibin alpha-subunit           gene, exon 1       M80826_at   M80826 Rat intestinal trefoil   790   787   70   −10   788.5   30   0.038047   −26.3           protein mRNA, complete           cds/cds = 17.262/           gb = M80826/gi = 207446/           ug = Rn.9960/len = 431       rc_AI639304_at   Rat mixed-tissue library   573   503   301   325   538   313   0.581784   −1.7             Rattus norvegicus  cDNA           clone rx00157 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AA799581_at   rc_AA799581 EST189078   429   462   209   179   445.5   194   0.435466   −2.3             Rattus norvegicus  cDNA, 3′           end/clone = RHEAC77/           clone_end = 3′/           gb = AA799581/gi = 2862536/           ug = Rn.6207/len = 569       rc_AA800211_at   rc_AA800211 EST189708   164   224   326   400   194   363   1.871134   1.9             Rattus norvegicus  cDNA, 3′           end/clone = RHEAM49/           clone_end = 3′/           gb = AA800211/gi = 2863166/           ug = Rn.6299/len = 740       rc_AA800549_at   rc_AA800549 EST190046   306   333   461   655   319.5   558   1.746479   1.7             Rattus norvegicus  cDNA, 3′           end/clone = RLUAB29/           clone_end = 3′/           gb = AA800549/gi = 2863504/           ug = Rn.22957/len = 491       rc_AA800749_at   rc_AA800749 EST190246   532   392   234   193   462   213.5   0.462121   −2.2             Rattus norvegicus  cDNA, 3′           end/clone = RLUAL02/           clone_end = 3′/           gb = AA800749/gi = 2863704/           ug = Rn.1897/len = 637       rc_AA859680_g_at   rc_AA859680 UI-R-E0-bs-d-   2002   1841   944   761   1921.5   852.5   0.443664   −2.3           12-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bs-d-12-0-           UI/clone_end = 3′/           gb = AA859680/gi = 2949200/           ug = Rn.22632/len = 437       rc_AA874874_at   rc_AA874874 UI-R-E0-ci-d-   761   632   1099   1322   696.5   1210.5   1.737976   1.7           12-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ci-d-12-0-           UI/clone_end = 3′/           gb = AA874874/gi = 2979822/           ug = Rn.3157/len = 513       rc_AA874919_at   rc_AA874919 UI-R-E0-ck-g-   541   490   216   226   515.5   221   0.42871   −2.3           09-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ck-g-09-0-           UI/clone_end = 3′/           gb = AA874919/gi = 2979867/           ug = Rn.3174/len = 542       rc_AA875127_g_at   rc_AA875127 UI-R-E0-bu-   395   382   208   199   388.5   203.5   0.52381   −1.9           d-05-0-UI.s2  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bu-d-05-0-           UI/clone_end = 3′/           gb = AA875127/gi = 2980075/           ug = Rn.18698/len = 579       rc_AA891690_at   rc_AA891690 EST195493   167   189   391   335   178   363   2.039326   2.0             Rattus norvegicus  cDNA, 3′           end/clone = RKIAF58/           clone_end = 3′/           gb = AA891690/gi = 3018569/           ug = Rn.22701/len = 446       rc_AA891940_at   rc_AA891940 EST195743   109   29   385   426   69   405.5   5.876812   5.9             Rattus norvegicus  cDNA, 3′           end/clone = RKIAI82/           clone_end = 3′/           gb = AA891940/gi = 3018819/           ug = Rn.3508/len = 523       rc_AA892378_g_at   rc_AA892378 EST196181   959   890   1732   1866   924.5   1799   1.945917   1.9             Rattus norvegicus  cDNA, 3′           end/clone = RKIAP70/           clone_end = 3′/           gb = AA892378/gi = 3019257/           ug = Rn.1298/len = 589       rc_AA944423_at   rc_AA944423 EST199922   435   376   255   200   405.5   227.5   0.561036   −1.8             Rattus norvegicus  cDNA, 3′           end/clone = REMAJ02/           clone_end = 3′/           gb = AA944423/gi = 3104339/           ug = Rn.6165/len = 670       rc_AA946384_at   rc_AA946384 EST201883   464   624   352   278   544   315   0.579044   −1.7             Rattus norvegicus  cDNA, 3′           end/clone = RLUBH49/           clone_end = 3′/           gb = AA946384/gi = 3106300/           ug = Rn.11301/len = 576       rc_AI102868_g_at   rc_AI102868 EST212157   1431   1441   702   953   1436   827.5   0.576253   −1.7             Rattus norvegicus  cDNA, 3′           end/clone = REMBT90/           clone_end = 3′/gb = AI102868/           ug = Rn.221/len = 489       rc_AI228599_at   rc_AI228599 EST225294   295   395   68   42   345   55   0.15942   −6.3             Rattus norvegicus  cDNA, 3′           end/clone = RBRCW95/           clone_end = 3′/gb = AI228599/           ug = Rn.3877/len = 572       rc_AI236484_at   rc_AI236484 EST233046   124   115   247   263   119.5   255   2.133891   2.1             Rattus norvegicus  cDNA, 3′           end/clone = ROVDG74/           clone_end = 3′/gb = AI236484/           ug = Rn.3924/len = 474       rc_H31351_at   rc_H31351 EST105310   437   382   265   188   409.5   226.5   0.553114   −1.8             Rattus norvegicus  cDNA, 3′           end/clone = RPCAH85/           clone_end = 3′/           gb = H31351/gi = 976768/           ug = Rn.14564/len = 352       S70803_g_at   S70803 clone p10.15   584   699   147   199   641.5   173   0.26968   −3.7           product [rats, osteosarcoma           ROS17/2.8, mRNA, 737 nt]       U01146_s_at   U01146 RRU01146  Rattus     432   367   586   799   399.5   692.5   1.733417   1.7             rattus  Sprague Dawley           nuclear orphan receptor           HZF-3 (HZF-3) mRNA,           complete cds       U14192complete_seq_at   U14192completeSeq  Rattus     311   292   163   168   301.5   165.5   0.548922   −1.8             norvegicus  general           vesicular transport factor           p115 mRNA, complete cds/           cds = 11.2890/           gb = U14192/gi = 538152/           ug = Rn.4746/len = 2891       X03347cds_g_at   X03347cds REMSVFBR   232   304   461   496   268   478.5   1.785448   1.8           FBR-murine osteosarcoma           provirus genome       X12554cds_s_at   X12554cds RNCOX6AH   269   222   401   470   245.5   435.5   1.773931   1.8           Rat mRNA for heart           cytochrome c oxidase           subunit Via       X63446_at   X63446  R. norvegicus     520   388   248   249   454   248.5   0.547357   −1.8           mRNA for fetuin/           cds = 31,1089/gb = X63446/           gi = 56139/ug = Rn.3880/           len = 1456                  
 
         [0192]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and 1 week withdrawal in the CeA. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 6                           CeA 1-Week Withdrawal to Control                                1-week                                   1-week   1-week   with-               1-week   with-   extinction   drawal   Mean   Mean       Fold       Probe set no.   Description   extinction   drawal   control   control   control   extinction   Ratio   change                    AB003753cds#1_at   AB003753cds#1  Rattus     373   366   82   126   369.5   104   0.281461   −3.6             norvegicus  genes for           high sulfur protein B2E           and high sulfur protein           B2F, complete cds       AB015433_s_at   AB015433  Rattus     269   253   609   524   261   566.5   2.170498   2.2             norvegicus  mRNA for 4F2           heavy chain (4F2hc),           complete cds       AB016160_g_at   AB016160  Rattus     414   318   148   150   366   149   0.407104   −2.5             norvegicus  mRNA for           GABAB receptor 1c,           complete cds       AF063302mRNA#3_s_at   AF063302mRNA#3  Rattus     421   395   140   −4   408   68   0.166667   −6.0             norvegicus  carnitine           palmitoyltransferase           Ibeta 1, carnitine           palmitoyltransferase           Ibeta 2, and carnitine           palmitoyltransferase           Ibeta 3 gene, nuclear           gene encoding mito-           chondrial proteins,           alternatively spliced           products, partial cds       AF064856_at   AF064856  Rattus  sp.   561   529   332   244   545   288   0.52844   −1.9           7acomp protein mRNA,           complete cds       AF081144_s_at   AF081144  Rattus     288   202   495   578   245   536.5   2.189796   2.2             norvegicus  CL1AA mRNA,           complete cds       D10853_at   D10853 RATATR Rat   240   226   119   115   233   117   0.502146   −2.0           mRNA for amidophos-           phoribosyltransferase       D13309_s_at   D13309 RATRDBPB Rat   626   625   348   359   625.5   353.5   0.565148   −1.8           mRNA for DNA-binding           protein B       D64085_at   D64085 RATORFA1 Rat   443   344   114   245   393.5   179.5   0.456163   −2.2           mRNA for fibroblast           growth factor FGF-5,           complete cds       D83538_g_at   D83538 Rat mRNA for   178   202   386   470   190   428   2.252632   2.3           230 kDa phosphatidyli-           nositol 4-kinase,           complete cds/           cds = 391.6516/gb = D83538/           gi = 1339965/ug = Rn.11015/           len = 6857       J00771_at   J00771 RATPRNASE Rat   262   238   −50   −13   250   −31.5   −0.126   7.9           pancreatic ribo-           nuclease mRNA       L07398_at   L07398 RATIGVCL   305   311   723   611   308   667   2.165584   2.2             Rattus norvegicus             (hybridoma 56R-3)           immunoglobulin re-           arranged gamma-chain           mRNA variable (V)           region, partial cds       L19699_at   L19699 Rat GTP-binding   331   276   657   711   303.5   684   2.253707   2.3           protein (ral B) mRNA,           complete cds/           cds = 64.684/gb = L19699/           gi = 310211/ug = Rn.4586/           len = 2074       L40364_f_at   L40364  Rattus      177   129   475   403   153   439   2.869281   2.9             norvegicus  MHC class           I RT1.O type - 149           processed pseudogene           mRNA/cds = UNKNOWN/           gb = L40364/gi = 992568/           ug = Rn.3577/len = 1602       M55050_at   M55050  Rattus norwegicus     533   378   231   237   455.5   234   0.513721   −1.9           interleukin-2 receptor           beta chain (p70/75)           mRNA, complete cds/           cds = 111,1724/           gb = M55050/gi = 204913/           ug = Rn.5832/len = 2598       M81639_at   M81639  Rattus norvegicus     292   316   474   592   304   533   1.753289   1.8           stannin mRNA/           cds = UNKNOWN/           gb = M81639/gi = 207078/           ug = Rn.6147/len = 2897       rc_AI639096_at   Rat mixed-tissue library   111   239   392   383   175   387.5   2.214286   2.2             Rattus norvegicus  cDNA           clone rx00904 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AI639391_at   Rat mixed-tissue library   982   1009   284   334   995.5   309   0.310397   −3.2             Rattus norvegicus  cDNA           clone rx02754 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AI638980_at   Rat mixed-tissue library   631   601   277   221   616   249   0.404221   −2.5             Rattus norvegicus  cDNA           clone rx03968 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AI639195_r_at   Rat mixed-tissue library   822   933   519   393   877.5   456   0.519658   −1.9             Rattus norvegicus  cDNA           clone rx04881 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AA799421_at   rc_AA799421 EST188918   359   319   479   675   339   577   1.702065   1.7             Rattus norvegicus             cDNA, 3′ end/           clone = RHEAA87/           clone_end = 3′/           gb = AA799421/gi = 2862376/           ug = Rn.19951/len = 570       rc_AA799449_g_at   rc_AA799449 EST188946   262   327   470   670   294.5   570   1.935484   1.9             Rattus norvegicus             cDNA, 3′ end/           clone = RHEAB19/           clone_end = 3′/           gb = AA799449/gi = 2862404/           ug = Rn.3286/len = 553       rc_AA799671_at   rc_AA799671 EST189168   421   529   249   297   475   273   0.574737   −1.7             Rattus norvegicus             cDNA, 3′ end/           clone = RHEAD82/           clone_end = 3′/           gb = AA799671/gi = 2862626/           ug = Rn.6219/len = 328       rc_AA799899_i_at   rc_AA799899 EST189396   4497   3805   6266   7851   4151   7058.5   1.700434   1.7             Rattus norvegicus             cDNA, 3′ end/           clone = RHEAG67/           clone_end = 3′/           gb = AA799899/gi = 2862854/           ug = Rn.5974/len = 505       rc_AA859680_g_at   rc_AA859680 UI-R-E0-bs-   731   959   2002   1841   845   1921.5   2.273964   2.3           d-12-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bs-d-12-           0-UI/clone_end = 3′/           gb = AA859680/gi = 2949200/           ug = Rn.22632/len = 437       rc_AA875054_at   rc_AA875054 UI-R-E0-   779   581   320   455   680   387.5   0.569853   −1.8           cb-e-04-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cb-e-04-           0-UI/clone_end = 3′/           gb = AA875054/gi = 2980002/           ug = Rn.24874/len = 485       rc_AA891438_g_at   rc_AA891438 EST195241   557   438   58   235   497.5   146.5   0.294472   −3.4             Rattus norvegicus             cDNA, 3′ end/           clone = RHEAU25/           clone_end = 3′/           gb = AA891438/gi = 3018317/           ug = Rn.22406/len = 397       rc_AA891690_at   rc_AA891690 EST195493   316   308   167   189   312   178   0.570513   −1.8             Rattus norvegicus             cDNA, 3′ end/           clone = RKIAF58/           clone_end = 3′/           gb = AA891690/gi =3018569/           ug = Rn.22701/len = 446       rc_AA892859_at   rc_AA892859 EST196662   236   225   −51   −31   230.5   −41   −0.17787   5.6             Rattus norvegicus             cDNA, 3′ end/           clone = RKIAY19/           clone_end = 3′/           gb = AA892859/gi = 3019738/           ug = Rn.8137/len = 568       rc_AA899106_at   rc_AA899106 UI-R-E0-cw-   550   698   170   185   624   177.5   0.284455   −3.5           d-04-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cw-d-04-           0-UI/clone_end = 3′/           gb = AA899106/gi = 3034460/           ug = Rn.6031/len = 523       rc_AA944422_at   rc_AA944422 EST199921   109   240   382   519   174.5   450.5   2.581662   2.6             Rattus norvegicus             cDNA, 3′ end/           clone = REMAJ01/           clone_end = 3′/           gb = AA944422/gi = 3104338/           ug = Rn.871/len = 641       rc_AI060085_s_at   rc_AI060085 UI-R-C1-Ii-   263   258   137   117   260.5   127   0.487524   −2.1           c-08-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C1-Ii-c-08-           0-UI/clone_end = 3′/           gb = AI060085/ug = Rn.9967/           len = 315       rc_AI138143_at   rc_AI138143 UI-R-C0-if-   219   210   119   101   214.5   110   0.512821   −2.0           e-07-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C0-if-e-07-           0-UI/clone_end = 3′/           gb = AI138143/ug = Rn.10708/           len = 343       rc_AI170212_s_at   rc_AI170212 EST216137   271   280   552   626   275.5   589   2.137931   2.1             Rattus norvegicus  cDNA, 3′           end/clone = RLUCF03/           clone_end = 3′/           gb = AI170212/gi = 3710252/           ug = Rn.11007/len = 322       rc_AI170268_at   rc_AI170268 EST216194   361   290   492   620   325.5   556   1.708141   1.7             Rattus norvegicus  cDNA, 3′           end/clone = RLUCG30/           clone_end = 3′/           gb = AI170268/gi = 3710308/           ug = Rn.1868/len = 577       rc_AI176488_at   rc_AI176488 EST220073   300   391   188   27   345.5   107.5   0.311143   −3.2             Rattus norvegicus  cDNA, 3′           end/clone = ROVBS47/           clone_end = 3′/gb = AI176488/           ug = Rn.9909/len = 650       rc_AI228599_at   rc_AI228599 EST225294   −79   −37   295   395   −58   345   −5.94828   at least             Rattus norvegicus  cDNA, 3′                               2-fold           end/clone = RBRCW95/           clone_end = 3′/gb = AI228599/           ug = Rn.3877/len = 572       rc_AI231519_at   rc_AI231519 EST228207   175   180   403   361   177.5   382   2.152113   2.2             Rattus norvegicus  cDNA, 3′           end/clone = REMDL26/           clone_end = 3′/gb = AI231519/           ug = Rn.6602/len = 482       Rc_H33651_at   rc_H33651 EST109846   406   309   216   189   357.5   202.5   0.566434   −1.8             Rattus norvegicus  cDNA, 3′           end/clone = RPNAV67/           clone_end = 3′/           gb = H33651/gi = 979068/           ug = Rn.14654/len = 447       U14414_at   U14414  Rattus norvegicus     281   294   126   129   287.5   127.5   0.443478   −2.3           P2x receptor mRNA,           complete cds/cds = 36,1454/           gb = U14414/gi = 558830/           ug = Rn.10991/len = 1831       U70270UTR#1_f_at   U70270UTR#1 RNMUD402   537   468   −153   66   502.5   −43.5   −0.08657   11.6             Rattus norvegicus  mud-4           mRNA, 3′ UTR       U75921UTR#1_at   U75921UTR#1 RNAPCBP3   412   388   122   181   400   151.5   0.37875   −2.6             Rattus norvegicus  APC           binding protein EB1           mRNA, 3′ untranslated           region, partial sequence       X03347cds_at   X03347cds REMSVFBR   463   513   252   117   488   184.5   0.378074   −2.6           FBR-murine osteosarcoma           provirus genome       X12554cds_s_at   X12554cds RNCOX6AH   544   449   269   222   496.5   245.5   0.494461   −2.0           Rat mRNA for heart           cytochrome c oxidase           subunit VIa       X15679_at   X15679 Rat mRNA for   707   595   371   362   651   366.5   0.56298   −1.8           preprotrypsinogen IV (EC           3.4.21.4)/cds = 14,757/           gb = X15679/gi = 56813/           ug = Rn.10387/len = 862       X60651mRNA_s_at   X60651mRNA RNSYNDCN   407   374   169   191   390.5   180   0.460948   −2.2           Rat mRNA for syndecan       X73579_at   X73579  R. norvegicus  CD23   −43   23   466   604   −10   535   −53.5   at least           mRNA/cds = 0.929/                               2-fold           gb = X73579/gi = 313672/           ug = Rn.10326/len = 1146                  
 
         [0193]     Genes that passed the filtering criteria outlined above for differential expression between 1 week withdrawal and its corresponding control in the CeA. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 7                           Core 1-Week Extinction to Control                    1-week   1-week   1-week   1-week                   extinction   extinction   extinction   extinction   Fold       Experiment   Description   control core   control core   core   core   change                    K02248cds_s_at   K02248cds RATSOM141 Rat   575   528   342   274   −1.8           somatostatin-14 gene, complete cds       M55534mRNA_s_at   M55534mRNA Rat alpha-crystallin B   167   264   416   414   1.8           chain mRNA, complete cds/           cds = UNKNOWN/gb = M55534/           gi = 203609/ug = Rn.832/len = 1247       Rc_AA894296_at   rc_AA894296 EST198099  Rattus     209   217   436   362   1.9             norvegicus  cDNA, 3′ end/           clone = RSPAW17/clone_end = 3′/           gb = AA894296/gi = 3021175/           ug = Rn.3760/len = 600       Rc_AI058941_s_at   rc_AI058941 UI-R-C1-Ir-b-07-0-UI.s1   222   −3   389   372   1.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-C1-Ir-b-07-0-UI/           clone_end = 3′/gb = AI058941/           ug = Rn.4231/len = 476       X15679_at   X15679 Rat mRNA for   353   365   201   120   −1.8           preprotrypsinogen IV (EC 3.4.21.4)/           cds = 14.757/gb = X15679/           gi = 56813/ug = Rn.10387/len = 862       X95990exon_s_at   X95990exon RNC5ARECP   645   544   328   360   −1.7             R. norvegicus  mRNA for C5a           anaphylatoxin receptor       Z11581_at   Z11581  R. norvegicus  mRNA for   683   724   357   460   −1.7           kainate receptor subunit (ka2)/           cds = 202.3141/gb = Z11581/gi = 56509/           ug = Rn.10053/len = 3702       U05013_at   U05013  Rattus norvegicus  Sprague-   209   241   48   53   4.4           Dawley heme oxygenase-2 non-           reducing isoform gene, complete cds/           cds = 177.1124/gb = U05013/           gi = 501034/ug = Rn.10241/len = 1815       M64785_g_at   M64785 RATVAS Rat vasopressin (VP)   200   211   116   110   1.8           mRNA                  
 
         [0194]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and its corresponding control in the nucleus accumbens core. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 8                           Core 1-Week Extinction to Withdrawal                    1-week   1-week   1-week   1-week                   withdrawal   withdrawal   extinction   extinction   Fold       Experiment   Description   B   A   B   A   change                    AF055714UTR#1_at   AF055714UTR#1  Rattus norvegicus     481   466   2   −17   −2.4           hypertension-regulated vascular factor-           1C-4 mRNA, 3′ UTR       AF102855_at   AF102855  Rattus norvegicus  synaptic   238   264   110   109   2           SAPAP-interacting protein Synamon           mRNA, complete cds       M11071_f_at   M11071 Rat MHC class I cell surface   1021   897   2204   1642   2.0           antigen mRNA/cds = 0,330/gb = M11071/           gi = 205414/ug = Rn.11168/len = 824       M25890_at   M25890 Rat somatostatin mRNA,   875   668   1269   1448   1.8           complete cds/cds = 60.410/gb = M25890/           gi = 207030/ug = Rn.540/len = 564       M92076_at   M92076 RATMGLURC Rat   256   359   709   668   2.2           metabotropic glutamate receptor 3           Mrna, primary transcript       M95591_g_at   M95591 RATSST  Rattus rattus  hepatic   472   494   141   235   −2.2           squalene synthetase mRNA, complete           cds       M96626_g_at   M96626 RAT plasma membrane CA2+−   206   222   96   76   2           ATPase isoform 3 mRNA, partial cds/           cds = 0.346/gb = M96626/gi = 203212/           ug = Rn.11053/len = 609       rc_AI638989_at   Rat mixed-tissue library  Rattus     168   135   451   368   2.0             norvegicus  cDNA clone rx01268 3′,           mRNA sequence [ Rattus norvegicus ]       rc_AA819776_f_at   rc_AA819776 UI-R-A0-ap-h-07-0-UI.s1   56   −42   471   384   2.1           UI-R-A0  Rattus norvegicus  cDNA clone           UI-R-A0-ap-h-07-0-UI 3′ similar to           gb|J04633|MUSHSP86A Mouse heat           shock protein 86 mRNA, complete cds,           and 28S ribosomal RNA, partial           sequence, mRNA sequence [ Rattus               norvegicus ]       rc_AA858621_g_at   rc_AA858621 UI-R-E0-bq-b-10-0-UI.s1   439   335   691   870   2.0             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bq-b-10-0-UI/           clone_end = 3′/gb = AA858621/           gi = 2948961/ug = Rn.3551/len = 550       rc_AA859520_at   rc_AA859520 UI-R-E0-br-b-02-0-UI.s1   230   297   535   507   2.0             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-br-b-02-0-UI/           clone_end = 3′/gb = AA859520/           gi = 2949040/ug = Rn.23034/len = 453       rc_AA859966_i_at   rc_AA859966 UI-R-E0-ca-g-03-0-UI.s1   −129   −223   5469   5453   27.3             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ca-g-03-0-UI/           clone_end = 3′/gb = AA859966/           gi = 2949486/ug = Rn.861/len = 392       rc_AA875103_at   rc_AA875103 UI-R-E0-cf-h-04-0-UI.s1   299   266   −20   −49   14             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cf-h-04-0-UI/           clone_end = 3′/gb = AA875103/           gi = 2980051/ug = Rn.22643/len = 606       rc_AA875131_at   rc_AA875131 UI-R-E0-bu-e-03-0-UI.s2   381   429   186   231   −1.9             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bu-e-03-0-UI/           clone_end = 3′/gb = AA875131/           gi = 2980079/ug = Rn.2801/len = 575       rc_AA891721_at   rc_AA891721 EST195524  Rattus     342   417   166   170   −1.9             norvegicus  cDNA, 3′ end/           clone = RKIAF94/clone_end = 3′/           gb = AA891721/gi = 3018600/           ug = Rn.14709/len = 454       rc_AA893065_at   rc_AA893065 EST196868  Rattus     225   254   516   489   2.1             norvegicus  cDNA, 3′ end/           clone = RKIBB69/clone_end = 3′/           gb = AA893065/gi = 3019944/           ug = Rn.13472/len = 410       rc_AA893612_at   rc_AA893612 EST197415  Rattus     517   514   942   919   1.8             norvegicus  cDNA, 3′ end/           clone = RPLAC57/clone_end = 3′/           gb = AA893612/gi = 3020491/           ug = Rn.14814/len = 265       rc_AA893870_g_at   rc_AA893870 EST197673  Rattus     46   62   308   316   6             norvegicus  cDNA, 3′ end/           clone = RPLAM86/clone_end = 3′/           gb = AA893870/gi = 3020749/           ug = Rn.11229/len = 417       rc_AA945054_s_at   rc_AA945054 EST200553  Rattus     449   573   801   975   1.7             norvegicus  cDNA, 3′ end/           clone = RLIAF82/clone_end = 3′/           gb = AA945054/ug = Rn.1055/len = 565       rc_AA955983_at   rc_AA955983 UI-R-E1-fb-e-12-0-UI.s1   579   704   351   398   −1.7             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E1-fb-e-12-0-UI/           clone_end = 3′/gb = AA955983/           ug = Rn.7854/len = 542       rc_AI008863_at   rc_AI008863 EST203314  Rattus     322   450   196   245   −1.7             norvegicus  cDNA, 3′ end/           clone = REMBE50/clone_end = 3′/           gb = AI008863/ug = Rn.1893/len = 401       rc_AI013194_at   rc_AI013194 EST207869  Rattus     217   251   584   531   2             norvegicus  cDNA, 3′ end/           clone = RSPBH90/clone_end = 3′/           gb = AI013194/ug = Rn.3506/len = 464       rc_AI014135_g_at   rc_AI014135 EST207690  Rattus     1499   1401   567   444   3             norvegicus  cDNA, 3′ end/           clone = RSPBF48/clone_end = 3′/           gb = AI014135/ug = Rn.4229/len = 410       rc_AI102103_at   rc_AI102103 EST211392  Rattus     1193   1211   698   655   −1.8             norvegicus  cDNA, 3′ end/           clone = RBRBY91/clone_end = 3′/           gb = AI102103/gi = 3706936/           ug = Rn.14991/len = 611       rc_AI172097_g_at   rc_AI172097 EST218092  Rattus     274   323   541   556   1.8             norvegicus  cDNA, 3′ end/           clone = RMUBU88/clone_end = 3′/           gb = AI172097/gi = 3712137/           ug = Rn.20418/len = 570       rc_H31982_at   rc_H31982 EST106584  Rattus     354   431   170   175   −2.0             norvegicus  cDNA, 3′ end/           clone = RPCBE17/clone_end = 3′/           gb = H31982/gi = 977399/ug = Rn.7138/           len = 363       U62897_at   U62897  Rattus norvegicus     183   216   344   435   1.9           carboxypeptidase D precursor (Cpd)           mRNA, complete cds/cds = 45.4181/           gb = U62897/gi = 2406562/           ug = Rn.4093/len = 4377       U67995_s_at   U67995  Rattus norvegicus  stearyl-CoA   1336   1291   780   591   −1.9           desaturase 2 mRNA, partial cds/           cds = 0.92/gb = U67995/gi = 1763026/           ug = Rn.10650/len = 315       U77931_at   U77931 RNU77931  Rattus norvegicus     836   912   2166   1850   2.3           unknown mRNA       X05472cds#2_at   X05472cds#2 RNREP24R Rat 2.4 kb   4218   4342   945   541   6           repeat DNA right terminal region       X06564_at   X06564 Rat mRNA for 140-kD NCAM   47   28   309   281   8           polypeptide/cds = 208.2784/           gb = X06564/gi = 56736/ug = Rn.11283/           len = 3170       X12744_at   X12744 Rat mRNA for c-erb-A thyroid   255   252   499   442   1.9           hormone receptor/cds = 0.1198/           gb = X12744/gi = 55931/ug = Rn.11307/           len = 1775       X15679_at   X15679 Rat mRNA for   377   403   120   201   −1.9           preprotrypsinogen IV (EC 3.4.21.4)/           cds = 14.757/gb = X15679/           gi = 56813/ug = Rn.10387/len = 862       X70667cds_at   X70667cds RRMC3RA  R. rattus  mRNA   221   249   426   508   2.0           for melanocortin-3 receptor       AFFX_rat_5S_rRNA_at   X83747  Rattus norvegicus  5S rRNA   348   357   154   146   2           gene (clone pRA5S2).                  
 
         [0195]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and 1 week withdrawal in the nucleus accumbens core. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 9                           Core 1-Week Withdrawal to Control                    1-week   1-week                           withdrawal   withdrawal   1-week   1-week               control   control   withdrawal   withdrawal   Fold       Experiment   Description   A   B   A   B   change                    AB008424_s_at   AB008424  Rattus norvegicus  mRNA for   376   453   180   153   −2.1           CYP2D3, complete cds       AF069525_at   AF069525  Rattus norvegicus  190 kDa   275   234   504   424   1.8           ankyrin isoform mRNA, complete cds/           cds = 84.5372/gb = AF069525/           gi = 3202045/ug = Rn.236/len = 6184       AF077354_g_at   AF077354  Rattus norvegicus  ischemia   61   81   244   251   3.5           responsive 94 kDa protein (irp94)           mRNA, complete cds       AJ005425_at   AJ005425 RNAJ5425  Rattus     86   22   373   394   1.9             norvegicus  mRNA for MEF2D protein       L07398_at   L07398 RATIGVCL  Rattus norvegicus     437   308   231   156   −1.7           (hybridoma 56R-3) immunoglobulin           rearranged gamma-chain mRNA           variable (V) region, partial cds       M80826_at   M80826 Rat intestinal trefoil protein   334   322   112   102   3.1           mRNA, complete cds/cds = 17.262/           gb = M80826/gi = 207446/           ug = Rn.9960/len = 431       Rc_AI639392_at   Rat mixed-tissue library  Rattus     291   393   96   89   −1.7             norvegicus  cDNA clone rx02714 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_AA875131_at   rc_AA875131 UI-R-E0-bu-e-03-0-UI.s2   201   260   429   381   1.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bu-e-03-0-UI/           clone_end = 3′/gb = AA875131/           gi = 2980079/ug = Rn.2801/len = 575       Rc_AA899106_at   rc_AA899106 UI-R-E0-cw-d-04-0-UI.s1   105   120   252   273   2.3             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cw-d-04-0-UI/           clone_end = 3′/gb = AA899106/           gi = 3034460/ug = Rn.6031/len = 523       Rc_AI230778_at   rc_AI230778 EST227473  Rattus     341   359   142   122   −1.8             norvegicus  cDNA, 3′ end/           clone = REMDB16/clone_end = 3′/           gb = AI230778/ug = Rn.3659/len = 560       Rc_AI230778_at   rc_AI230778 EST227473  Rattus     359   341   122   142   2.7             norvegicus  cDNA, 3′ end/           clone = REMDB16/clone_end = 3′/           gb = AI230778/ug = Rn.3659/len = 560       U38180_at   U38180  Rattus norvegicus  reduced   124   110   277   253   2.3           folate carrier membrane glycoprotein           mRNA, complete cds/cds = 248.1786/           gb = U38180/gi = 1022954/ug =Rn.9042/           len = 2410       U70268UTR#1_at   U70268UTR#1 RNMUD702  Rattus     670   600   317   363   −1.9             norvegicus  mud-7 mRNA, 3′ UTR       X56729mRNA_at   X56729mRNA RSCALPST Rat mRNA   324   322   64   64   5.1           for calpastatin                  
 
         [0196]     Genes that passed the filtering criteria outlined above for differential expression between 1 week withdrawal and its corresponding control in the nucleus accumbens core. Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 10                           mPFC F Id Change 1-Week Extinction to Control                    1-week   1-week                           withdrawal   withdrawal   1-week   1-week               control   control   withdrawal   withdrawal   Fold       Experiment   Description   A   B   A   B   change                    AB002393_at   AB002393  Rattus norvegicus  mRNA   230   198   −38   −45   −10.7           for histidase, partial cds       AB012234_g_at   AB012234  Rattus norvegicus  mRNA   719   751   440   358   −1.8           for NF1-X1, partial cds/cds = 0.535/           gb = AB012234/gi = 2982735/           ug = Rn.9647/len = 601       AF050663UTR#1_at   AF050663UTR#1  Rattus norvegicus     492   471   200   173   −2.6           activity and neurotransmitter-induced           early gene 11 (ania-11) mRNA,           3′ UTR       AF081204_s_at   AF081204  Rattus norvegicus  small   414   402   220   212   −1.9           intestine sodium dependent           multivitamin transporter (SMVT)           mRNA, complete cds       AF102854_at   AF102854  Rattus norvegicus     458   430   190   123   −2.2           membrane-associated guanylate           kinase-interacting protein 2 Maguin-2           mRNA, complete cds       AJ005113_g_at   AJ005113 RNAJ5113  Rattus     447   469   232   258   −1.9             norvegicus  mRNA for SMC-protein           Molecular characterization of a rat           heterochromatin associated SMC-           protein       AJ011115_at   AJ011115 RNO011115  Rattus     425   315   83   129   −1.9             norvegicus  mRNA for endothelial nitric           oxide synthase, 5′ region, partial       AJ012603UTR#1_at   AJ012603UTR#1 RNO012603  Rattus     520   442   211   237   −2.1             norvegicus  mRNA for TNF-alpha           converting enzyme (TACE)       D00512_g_at   D00512 RATACAL  Rattus  sp. mRNA   464   365   203   173   −2.1           for mitochondrial acetoacetyl-CoA           thiolase precursor, complete cds       D30040_at   D30040 Rat mRNA for RAC protein   206   229   383   450   1.9           kinase alpha, complete cds/           cds = 42.1484/gb = D30040/gi = 485402/           ug = Rn.11422/len = 1617       E01415cds_s_at   E01415cds cDNA encoding rat   975   687   501   460   −1.7           glutathione S transferase       J02592_s_at   J02592 Rat glutathione S-transferase   1022   746   265   347   −2.9           Y-b subunit mRNA, 3′ end/cds = 0.560/           gb = J02592/gi = 204498/ug = Rn.625/           len = 909       J05155_at   J05155 Rat phospholipase C type IV   228   222   72   88   −2.8           mRNA, complete cds/cds = 200.3997/           gb = J05155/gi = 206242/ug = Rn.9751/           len = 4321       K01701_at   K01701 Rat oxytocin/neurophysin   150   162   418   508   2.3           (Oxt) gene, complete gene, complete           cds/cds = 41.418/gb = K01701/           gi = 205899/ug = Rn.11315/len = 530       L37971mRNA_at   L37971 mRNA RATTCRAP  Rattus     349   340   171   203   −1.7             norvegicus  T-cell receptor alpha-chain           mRNA       L38482_at   L38482  Rattus norvegicus  serine   253   355   687   603   2.1           protease gene, complete cds/           cds = 0.401/gb = L38482/gi = 1020080/           ug = Rn.2427/len = 402       M22756_at   M22756 Rat 24-kDa subunit of   1247   1029   604   651   −1.8           mitochondrial NADH dehydrogenase           mRNA, 3′ end/cds = 0.725/           gb = M22756/gi = 205627/ug = Rn.11092/           len = 771       M25804_g_at   M25804 Rat Rev-ErbA-alpha protein   58   161   418   365   2.0           mRNA, complete cds/cds = 501.2027/           gb = M25804/gi = 514963/ug = Rn.10105/           len = 2297       M27886exon_g_at   M27886exon RAT6PF2KFR  Rattus     308   301   72   72   −4.2             norvegicus  bifunctional enzyme 6-           phosphofructo-2-kinase/fructose           2,6-bisphosphatase (6-PF2-K/           Fru-2,6-P-2-ase) gene, exon 1       M31032cds#1_s_at   M31032cds#1 RATCRP01 Rat   426   354   180   176   −2.0           contiguous repeat polypeptides (CRP)           mRNA, complete cds       M32061_at   M32061 Rat alpha-2B-adrenergic   158   236   508   481   2.3           receptor (RNG-alpha-2) mRNA,           complete cds/cds = 365.1726/           gb = M32061/gi = 202589/ug = Rn.10296/           len = 2319       M76535cds_at   M76535cds RATCXN40A Rat gap   734   746   353   278   −2.4           junction structural protein, connexin           (CXN-40) gene, complete cds       M77245_at   M77245  R. norvegicus  beta′-chain   23   162   415   512   2.3           clathrin associated protein complex           AP-1 mRNA, complete cds/cds = 39.2888/           gb = M77245/gi = 203112/ug = Rn.9466/           len = 3663       M77246_at   M77246  R. norvegicus  beta-chain   585   580   1166   1215   2.0           clathrin associated protein complex           AP-2 mRNA, complete cds/           cds = 139.2994/gb = M77246/gi = 203114/           ug = Rn.1050/len = 5402       M97662_at   M97662  Rattus norvegicus  beta-   406   425   204   140   −2.1           alanine synthase mRNA, complete           cds/cds = 33.1214/gb = M97662/           gi = 203105/ug = Rn.11110/len = 1420       rc_AI639272_at   Rat mixed-tissue library  Rattus     248   261   55   73   −4.0             norvegicus  cDNA clone rx03958 3′,           mRNA sequence [ Rattus norvegicus ]       rc_AI639313_at   Rat mixed-tissue library  Rattus     576   564   191   154   −3.3             norvegicus  cDNA clone rx04777 3′,           mRNA sequence [ Rattus norvegicus ]       rc_AI639195_r_at   Rat mixed-tissue library  Rattus     -73   84   847   1043   4.7             norvegicus  cDNA clone rx04881 3′,           mRNA sequence [ Rattus norvegicus ]       rc_AA684641_at   rc_AA684641 EST104995  Rattus     135   197   356   348   1.8             norvegicus  cDNA, 3′ end/           clone = RPCAE71/clone_end = 3′/           gb = AA684641/gi = 2671239/           ug = Rn.14675/len = 249       rc_AA799525_at   rc_AA799525 EST189022  Rattus     682   583   371   370   −1.7             norvegicus  cDNA, 3′ end/           clone =RHEAC13/clone_end = 3′/           gb = AA799525/gi = 2862480/           ug =Rn.1099/len = 573       rc_AA799531_g_at   rc_AA799531 EST189028  Rattus     586   471   247   329   −1.8             norvegicus  cDNA, 3′ end/           clone = RHEAC22/clone_end = 3′/           gb = AA799531/gi = 2862486/           ug =Rn.6198/len = 570       rc_AA818152_f_at   rc_AA818152 UI-R-A0-am-b-09-0-   6678   7495   3932   4179   −1.7           UI.s1  Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-A0-am-b-09-0-UI/           clone_end = 3′/gb = AA818152/           gi = 2888032/ug = Rn.16465/len = 117       rc_AA818226_s_at   rc_AA818226 UI-R-A0-ah-g-06-0-   5530   4813   2582   3212   −1.8           UI.s1  Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-A0-ah-g-06-0-UI/           clone_end = 3′/gb = AA818226/           gi = 2888106/ug = Rn.2528/len = 609       rc_AA851403_g_at   rc_AA851403 EST194171  Rattus     1728   1725   781   1074   −1.9             norvegicus  cDNA, 3′ end/           clone = RPLAG17/clone_end = 3′/           gb = AA851403/gi = 2938943/           ug = Rn.3383/len = 393       rc_AA851403_at   rc_AA851403 EST194171  Rattus     291   296   131   127   −2.3             norvegicus  cDNA, 3′ end/           clone = RPLAG17/clone_end = 3′/           gb = AA851403/gi = 2938943/           ug = Rn.3383/len = 393       rc_AA852004_s_at   rc_AA852004 EST194773  Rattus     780   722   1393   1184   1.7             norvegicus  cDNA, 3′ end/           clone = RSPAP38/clone_end = 3′/           gb = AA852004/gi = 2939544/           ug = Rn.2204/len = 368       rc_AA859299_at   rc_AA859299 UI-R-E0-cj-b-02-0-UI.s1   309   302   721   553   2.1             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cj-b-02-0-UI/           clone_end = 3′/gb = AA859299/           gi = 2948650/ug = Rn.9517/len = 529       rc_AA859837_g_at   rc_AA859837 UI-R-E0-cc-g-09-0-   3301   2600   1719   1534   −1.8           UI.s1  Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cc-g-09-0-UI/           clone_end = 3′/gb = AA859837/           gi = 2949357/ug = Rn.24783/len = 486       rc_AA859922_at   rc_AA859922 UI-R-E0-cg-c-04-0-   657   601   253   321   −2.2           UI.s1  Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cg-c-04-0-UI/           clone_end = 3′/gb = AA859922/           gi = 2949442/ug = Rn.819/len = 373       rc_AA866477_at   rc_AA866477 UI-R-E-br-h-03-0-UI.s1   1136   1364   601   702   −1.9             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-br-h-03-0-UI/           clone_end = 3′/gb = AA866477/           gi = 2961938/ug = Rn.2026/len = 488       rc_AA875420_at   rc_AA875420 UI-R-E0-cs-e-08-0-UI.s1   291   339   20   47   −9.4             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cs-e-08-0-UI/           clone_end = 3′/gb = AA875420/           gi = 2980368/ug = Rn.21413/len = 499       rc_AA892006_at   rc_AA892006 EST195809  Rattus     −157   −160   510   449   23.96             norvegicus  cDNA, 3′ end/           clone = RKIAK60/clone_end = 3′/           gb = AA892006/gi = 3018885/           ug = Rn.11519/len = 443       rc_AA892800_at   rc_AA892800 EST196603  Rattus     −203   −165   390   313   1.8             norvegicus  cDNA, 3′ end/           clone = RKIAX43/clone_end = 3′/           gb = AA892800/gi = 3019679/           ug = Rn.3609/len = 493       rc_AA894296_at   rc_AA894296 EST198099  Rattus     222   252   457   573   2.2             norvegicus  cDNA, 3′ end/           clone = RSPAW17/clone_end = 3′/           gb = AA894296/gi = 3021175/           ug = Rn.3760/len = 600       rc_AA899106_at   rc_AA899106 UI-R-E0-cw-d-04-0-UI.s1   482   459   292   214   −1.9             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cw-d-04-0-UI/           clone_end = 3′/gb = AA899106/           gi = 3034460/ug = Rn.6031/len = 523       rc_AA899253_at   rc_AA899253 UI-R-E0-cz-g-07-0-UI.s1   832   904   401   537   −1.9             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cz-g-07-0-UI/           clone_end = 3′/gb = AA899253/           gi = 3034607/ug = Rn.9560/len = 410       rc_AA945152_s_at   rc_AA945152 EST200651  Rattus     22042   30447   12827   15228   −1.9             norvegicus  cDNA, 3′ end/           clone = RLIAH24/clone_end = 3′/           gb = AA945152/ug = Rn.4241/len = 777       rc_AI009191_at   rc_AI009191 EST203642  Rattus     441   615   981   821   1.7             norvegicus  cDNA, 3′ end/           clone = REMBK67/clone_end = 3′/           gb = AI009191/ug = Rn.2432/len = 484       rc_AI058941_s_at   rc_AI058941 UI-R-C1-Ir-b-07-0-UI.s1   570   562   214   252   −2.4             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-C1-Ir-b-07-0-UI/           clone_end = 3′/gb = AI058941/           ug = Rn.4231/len = 476       rc_AI072770_s_at   rc_AI072770 UI-R-Y0-md-g-02-0-UI.s1   330   258   462   566   1.7             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-Y0-md-g-02-0-UI/           clone_end = 3′/gb = AI072770/           ug = Rn.4550/len = 333       rc_AI103396_g_at   rc_AI103396 EST212685  Rattus     26045   26104   16586   12308   −1.8             norvegicus  cDNA, 3′ end/           clone = REMCB47/clone_end = 3′/           gb = AI103396/gi = 3707945/           ug = Rn.221/len = 443       rc_AI137043_at   rc_AI137043 UI-R-C2p-oj-c-01-0-UI.s1   371   442   95   35   −2.0             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-C2p-oj-c-01-0-UI/           clone_end = 3′/gb = AI137043/           ug = Rn.22168/len = 436       rc_AI137856_s_at   rc_AI137856 UI-R-C0-ik-a-10-0-UI.s1   510   469   212   219   −2.3             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-C0-ik-a-10-0-UI/           clone_end = 3′/gb = AI137856/           ug = Rn.11359/len = 384       rc_AI176307_at   rc_AI176307 EST219889  Rattus     1777   2176   952   906   −2.1             norvegicus  cDNA, 3′ end/           clone = ROVBP82/clone_end = 3′/           gb = AI176307/ug = Rn.10427/len = 678       rc_AI176621_at   rc_AI176621 EST220210  Rattus     400   289   196   117   −1.7             norvegicus  cDNA, 3′ end/           clone = ROVBU65/clone_end = 3′/           gb = AI176621/ug = Rn.1979/len = 620       rc_AI177503_at   rc_AI177503 EST221135  Rattus     276   273   520   454   1.8             norvegicus  cDNA, 3′ end/           clone = RPLCA81/clone_end = 3′/           gb = AI177503/ug = Rn.11066/len = 575       rc_AI232012_at   rc_AI232012 EST228700  Rattus     1062   957   575   590   −1.7             norvegicus  cDNA, 3′ end/           clone = RHECR46/clone_end = 3′/           gb = AI232012/ug = Rn.1128/len = 586       rc_AI232321_at   rc_AI232321 EST229009  Rattus     312   333   177   173   −1.8             norvegicus  cDNA, 3′ end/           clone =RKICA22/clone_end = 3′/           gb = AI232321/ug = Rn.24630/len = 590       rc_AI234060_s_at   rc_AI234060 EST230748  Rattus     119   111   322   302   2.71             norvegicus  cDNA, 3′ end/           clone = RLUCU63/clone_end = 3′/           gb = AI234060/ug = Rn.11372/len = 363       S74801_s_at   S74801 H(+)-K(+)-ATPase alpha-   238   239   101   78   −2.7           subunit [rats, Sprague-Dawley,           kidney, mRNA Partial, 1361 nt]       U16025_at   U16025  Rattus norvegicus  class Ib   470   442   267   211   −1.9           RT1 mRNA, complete cds/cds = 0.1019/           gb = U16025/gi = 717092/ug = Rn.19044/           len = 1311       U23769_at   U23769  Rattus norvegicus  CLP36   172   162   285   284   1.7           (clp36) mRNA, complete cds/           cds = 66.1049/gb = U23769/gi = 1020150/           ug = Rn.11170/len = 1392       U32575_g_at   U32575 RNU32575  Rattus norvegicus     364   360   32   38   −1.8           (rsec6) mRNA, complete cds       U56261_s_at   U56261 RNU56261  Rattus norvegicus     122   144   300   303   2.27           SNAP-25a mRNA, partial cds       U70270UTR#1_f_at   U70270UTR#1 RNMUD402  Rattus     550   516   340   270   −1.7             norvegicus  mud-4 mRNA, 3′ UTR       U72995_at   U72995  Rattus norvegicus  Rab3   273   248   579   497   2.1           GDP/GTP exchange protein mRNA,           complete cds/cds = 191.4999/           gb = U72995/gi =1947049/ug = Rn.9786/           len = 5249       U89745_at   U89745  Rattus norvegicus  unknown   1075   1106   654   587   −1.8           protein mRNA, partial cds/cds = 0.293/           gb = U89745/gi = 1895082/ug = Rn.10720/           len = 1114       X53581cds#5_f_at   X53581cds#5 RNLINED  R. norvegicus     1225   1155   2071   2773   2.0           long interspersed repetitive DNA           containing 7 ORF&#39;s       X69903_at   X69903  R. norvegicus  mRNA for   491   377   118   146   −2.2           interleukin 4 receptor/cds = 9.2411/           gb = X69903/gi = 56390/ug = Rn.10471/           len = 2450       Y17048_g_at   Y17048 RNCALDE  Rattus norvegicus     492   465   912   916   1.91           mRNA for caldendrin       Z50052_at   Z50052  R. norvegicus  mRNA for C4BP   220   226   45   61   −4.2           beta chain protein/cds = 265.1041/           gb = Z50052/gi = 899381/ug = Rn.11151/           len = 1091                  
 
         [0197]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and its corresponding control in the medial prefrontal cortex (mPFC). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 11                           mPFC Fold Change 1-Week Extinction to Withdrawal                    1-week   1-week   1-week   1-week                   withdrawal   withdrawal   extinction   extinction   Fold       Experiment   Description   B   A   B   A   change                    AB004559_at   AB004559  Rattus norvegicus  mRNA for   34   109   393   431   −2.1           multispecific organic anion transporter,           complete cds/cds = 275.1930/           gb = AB004559/gi = 2361034/           ug = Rn.11113/len = 2221       AF020618_g_at   AF020618  Rattus norvegicus     371   416   205   139   1.9           progression elevated gene 3 protein           mRNA, complete cds       AF044201_at   AF044201  Rattus norvegicus  neural   1152   974   636   546   1.8           membrane protein 35 mRNA, complete           cds       AF051526_at   AF051526  Rattus norvegicus  class A   249   242   98   119   −2.3           calcium channel variant riA-I (BCCA1)           mRNA, partial cds/cds = 0.2375/           gb = AF051526/gi = 2961609/           ug = Rn.11281/len = 2427       AF076183_at   AF076183  Rattus norvegicus  cytosolic   405   319   215   177   1.7           sorting protein PACS-1a (PACS-1)           mRNA, complete cds       AF091566_f_at   AF091566  Rattus norvegicus  isolate   303   407   9   −44   1.8           HTF-SP1 olfactory receptor mRNA,           partial cds       AF102854_at   AF102854  Rattus norvegicus     379   395   123   190   1.9           membrane-associated guanylate           kinase-interacting protein 2 Maguin-2           mRNA, complete cds       AFFX_Rat_beta-   V01217 Rat gene encoding cytoplasmic   1708   2336   992   1100   1.9       actin_5_at   beta-actin (_5, _M, _3 represent           transcript regions 5 prime, Middle, and           3 prime respectively)       AJ005113_at   AJ005113 RNAJ5113  Rattus     299   384   103   195   1.7             norvegicus  mRNA for SMC-protein           Molecular characterization of a rat           heterochromatin associated SMC-           protein       AJ005394_at   AJ005394 RNJ005394  Rattus     355   309   103   96   −3.3             norvegicus  mRNA for collagen alpha 1           type V       AJ011005_at   AJ011005 RNO011005  Rattus     848   988   460   359   2.2             norvegicus  mRNA for Ptx3 protein       D00512_g_at   D00512 RATACAL  Rattus  sp. mRNA   386   470   173   203   2.1           for mitochondrial acetoacetyl-CoA           thiolase precursor, complete cds       D10757_at   D10757 RATPRORR12 Rat mRNA for   444   484   230   246   −2.0           proteasome subunit R-RING12,           complete cds       D13212_s_at   D13212 RATNMDARC Rat mRNA for   484   499   251   280   −1.9           N-methyl-D-aspartate receptor subunit           (NMDAR2C)       D14819_g_at   D14819 RATCBPP23B Rat mRNA for   679   890   484   392   1.8           calcium-binding protein P23k beta,           partial cds       D30734_at   D30734 RATGAP1M Rat mRNA for   353   383   220   208   1.7           Ras GTPase-activating protein,           complete cds       J02669_s_at   J02669 Rat cytochrome P-450a (3-   858   1000   539   530   1.7           methylchlanthrene-inducible; with high           testosterone 7-alpha activity), mRNA,           complete cds/cds = 19.1497/gb = J02669/           gi = 203766/ug = Rn.10904/len = 1687       J05499_at   J05499  Rattus norvegicus  L-glutamine   216   215   114   128   −1.8           amidohydrolase mRNA, complete cds/           cds = 131.1738/gb = J05499/gi = 1196813/           ug = Rn.10202/len = 2225       K01701_at   K01701 Rat oxytocin/neurophysin (Oxt)   150   131   508   418   −2.3           gene, complete gene, complete cds/           cds = 41.418/gb = K01701/gi = 205899/           ug = Rn.11315/len = 530       L07398_at   L07398 RATIGVCL  Rattus norvegicus     189   148   449   527   −2.4           (hybridoma 56R-3) immunoglobulin           rearranged gamma-chain mRNA           variable (V) region, partial cds       L38482_at   L38482  Rattus norvegicus  serine   290   360   603   687   −2.0           protease gene, complete cds/cds = 0.401/           gb = L38482/gi = 1020080/ug = Rn.2427/           len = 402       M11071_f_at   M11071 Rat MHC class I cell surface   3217   3367   792   959   −3.8           antigen mRNA/cds = 0.330/gb = M11071/           gi = 205414/ug = Rn.11168/len = 824       M20721_f_at   M20721 RATPRPA Rat proline-rich   282   281   129   128   −2.2           protein (PRP-1) mRNA, partial cds       M25804_g_at   M25804 Rat Rev-ErbA-alpha protein   175   138   365   418   −2.0           mRNA, complete cds/cds = 501.2027/           gb = M25804/gi = 514963/ug = Rn.10105/           len = 2297       M27886exon_g_at   M27886exon RAT6PF2KFR  Rattus     223   215   72   72   −3.0             norvegicus  bifunctional enzyme           6-phosphofructo-2-kinase/fructose           2,6-bisphosphatase (6-PF2-K/Fru-2,6-           P-2-ase) gene, exon 1       M31018_f_at   M31018  Rattus norvegicus  MHC class I   474   454   232   156   2.1           RT1.Aa alpha-chain precursor mRNA,           complete cds/cds = 9.1124/gb = M31018/           gi = 1877415/ug = Rn.3577/len = 1590       M77809_at   M77809 Rat betaglycan mRNA,   378   339   117   120   −3.0           complete cds/cds = 334.2895/gb = M77809/           gi = 203137/ug = Rn.9953/len = 3931       Rc_AA799467_at   rc_AA799467 EST188964  Rattus     413   486   292   218   1.8             norvegicus  cDNA, 3′ end/           clone = RHEAB38/clone_end = 3′/           gb = AA799467/gi = 2862422/ug = Rn.4036/           len = 568       Rc_AA799792_at   rc_AA799792 EST189289  Rattus     101   92   261   291   2.9             norvegicus  cDNA, 3′ end/clone = RHEAF41/           clone_end = 3′/gb = AA799792/           gi = 2862747/ug = Rn.7461/len = 615       Rc_AA799964_at   rc_AA799964 EST189461  Rattus     17   3   309   270   14.5             norvegicus  cDNA, 3′ end/clone = RHEAH66/           clone_end = 3′/gb = AA799964/           gi = 2862919/ug = Rn.6261/len = 452       Rc_AA800005_at   rc_AA800005 EST189502  Rattus     328   315   701   636   2.1             norvegicus  cDNA, 3′ end/clone = RHEAI20/           clone_end = 3′/gb = AA800005/           gi = 2862960/ug = Rn.1465/len = 628       Rc_AA800250_at   rc_AA800250 EST189747  Rattus     708   567   912   1264   −1.7             norvegicus  cDNA, 3′ end/clone = RHEAM94/           clone_end = 3′/gb = AA800250/           gi = 2863205/ug = Rn.3593/len = 666       Rc_AA800604_g_at   rc_AA800604 EST190101  Rattus     413   396   159   −18   2.0             norvegicus  cDNA, 3′ end/clone = RLUAB65/           clone_end = 3′/gb = AA800604/           gi = 2863559/ug = Rn.8590/len = 579       Rc_AA800737_at   rc_AA800737 EST190234  Rattus     219   206   430   322   −1.8             norvegicus  cDNA, 3′ end/clone = RLUAK84/           clone_end = 3′/gb = AA800737/           gi = 2863692/ug = Rn.6628/len = 626       Rc_AA851403_at   rc_AA851403 EST194171  Rattus     309   328   131   127   −2.5             norvegicus  cDNA, 3′ end/clone = RPLAG17/           clone_end = 3′/gb = AA851403/           gi = 2938943/ug = Rn.3383/len = 393       Rc_AA859585_at   rc_AA859585 UI-R-E0-bv-d-05-0-UI.s1   471   544   176   262   2.2             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bv-d-05-0-UI/           clone_end = 3′/gb = AA859585/           gi = 2949105/ug = Rn.24950/len = 516       Rc_AA859722_at   rc_AA859722 UI-R-E0-bx-h-09-0-UI.s1   459   381   −1   5   −21.0             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bx-h-09-0-UI/           clone_end = 3′/gb = AA859722/           gi = 2949242/ug = Rn.70/len = 460       Rc_AA859922_at   rc_AA859922 UI-R-E0-cg-c-04-0-UI.s1   615   712   321   253   2.3             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cg-c-04-0-UI/           clone_end = 3′/gb = AA859922/           gi = 2949442/ug = Rn.819/len = 373       Rc_AA874919_at   rc_AA874919 UI-R-E0-ck-g-09-0-UI.s1   224   221   365   428   −1.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ck-g-09-0-UI/           clone_end = 3′/gb = AA874919/           gi = 2979867/ug = Rn.3174/len = 542       Rc_AA875411_s_at   rc_AA875411 UI-R-E0-cs-b-11-0-UI.s1   115   191   425   422   −2.1             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cs-b-11-0-UI/           clone_end = 3′/gb = AA875411/           gi = 2980359/ug = Rn.2911/len = 423       Rc_AA892006_at   rc_AA892006 EST195809  Rattus     −59   −76   510   449   24.0             norvegicus  cDNA, 3′ end/           clone = RKIAK60/clone_end = 3′/           gb = AA892006/gi = 3018885/           ug =Rn.11519/len = 443       Rc_AA892179_at   rc_AA892179 EST195982  Rattus     210   198   421   358   −1.9             norvegicus  cDNA, 3′ end/           clone = RKIAN31/clone_end = 3′/           gb = AA892179/gi = 3019058/           ug = Rn.9031/len = 428       Rc_AA892800_at   rc_AA892800 EST196603  Rattus     35   −350   313   390   −1.8             norvegicus  cDNA, 3′ end/clone = RKIAX43/           clone_end = 3′/gb = AA892800/           gi = 3019679/ug = Rn.3609/len = 493       Rc_AA892801_g_at   rc_AA892801 EST196604  Rattus     497   658   277   354   1.8             norvegicus  cDNA, 3′ end/clone = RKIAX44/           clone_end = 3′/gb = AA892801/           gi = 3019680/ug = Rn.3610/len = 528       Rc_AA892828_at   rc_AA892828 EST196631  Rattus     343   240   444   551   −1.7             norvegicus  cDNA, 3′ end/clone = RKIAX75/           clone_end = 3′/gb = AA892828/           gi = 3019707/ug = Rn.2273/len = 626       Rc_AA893210_at   rc_AA893210 EST197013  Rattus     −20   28   329   361   17.3             norvegicus  cDNA, 3′ end/clone = RKIBD55/           clone_end = 3′/gb = AA893210/           gi = 3020089/ug = Rn.11141/len = 608       Rc_AI009191_at   rc_AI009191 EST203642  Rattus     512   542   821   981   −1.7             norvegicus  cDNA, 3′ end/clone = REMBK67/           clone_end = 3′/gb = AI009191/           ug = Rn.2432/len = 484       Rc_AI013993_at   rc_AI013993 EST207548  Rattus     279   248   100   102   −2.6             norvegicus  cDNA, 3′ end/clone = RSPBC95/           clone_end = 3′/gb = AI013993/           ug = Rn.221/len = 514       Rc_AI014094_g_at   rc_AI014094 EST207649  Rattus     374   335   195   187   1.8             norvegicus  cDNA, 3′ end/clone = RSPBE87/           clone_end = 3′/gb = AI014094/           ug = Rn.221/len = 569       Rc_AI101320_at   rc_AI101320 EST210609  Rattus     368   341   119   125   −2.9             norvegicus  cDNA, 3′ end/clone = RBRBL38/           clone_end = 3′/gb = AI101320/           ug = Rn.22459/len = 616       Rc_AI137856_s_at   rc_AI137856 UI-R-C0-ik-a-10-0-UI.s1   394   392   212   219   −1.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-C0-ik-a-10-O-UI/           clone_end = 3′/gb = AI137856/           ug = Rn.11359/len = 384       Rc_AI172097_g_at   rc_AI172097 EST218092  Rattus     533   441   123   200   2.4             norvegicus  cDNA, 3′ end/clone = RMUBU88/           clone_end = 3′/gb = AI172097/           gi = 3712137/ug = Rn.20418/len = 570       Rc_AI176307_at   rc_AI176307 EST219889  Rattus     1840   1824   906   952   −2.0             norvegicus  cDNA, 3′ end/clone = ROVBP82/           clone_end = 3′/gb = AI176307/           ug = Rn.10427/len = 678       Rc_AI231213_g_at   rc_AI231213 EST227901  Rattus     70   45   213   216   3.7             norvegicus  cDNA, 3′ end/clone = REMDH23/           clone_end = 3′/gb = AI231213/           ug = Rn.3022/len = 582       Rc_AI231472_s_at   rc_AI231472 EST228160  Rattus     160   171   384   349   2.2             norvegicus  cDNA, 3′ end/clone = REMDK57/           clone_end = 3′/gb = AI231472/           ug = Rn.2953/len = 549       Rc_AI639197_at   Rat mixed-tissue library  Rattus     706   904   379   388   2.1             norvegicus  cDNA clone rx02020 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_AI639236_at   Rat mixed-tissue library  Rattus     642   653   232   280   −2.5             norvegicus  cDNA clone rz00757 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_AI639313_at   Rat mixed-tissue library  Rattus     581   667   191   154   3.1             norvegicus  cDNA clone rx04777 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_H31420_at   rc_H31420 EST105436  Rattus     649   751   1229   1569   −2.0             norvegicus  cDNA, 3′ end/clone = RPCAJ34/           clone_end = 3′/gb = H31420/           gi = 976837/ug = Rn.8443/len = 312       S54212_at   S54212 ciliary neurotrophic factor   302   414   205   209   1.7           receptor alpha component [rats, brain,           mRNA, 1332 nt]       U20283_at   U20283  Rattus norvegicus  syntaxin   206   149   456   442   −2.2           binding protein Munc18-2 mRNA,           complete cds/cds = 6.1790/gb = U20283/           gi = 1022680/ug = Rn.10121/len = 2118       U35774_at   U35774  Rattus norvegicus  cytosolic   524   396   245   284   1.7           branch chain aminotransferase mRNA,           complete cds/cds = 62.1297/gb = U35774/           gi = 1173633/ug = Rn.8273/len = 1370       U36773_at   U36773 RNU36773  Rattus norvegicus     134   143   411   549   −2.4           glycerol-3-phosphate acyltransferase           mRNA, nuclear gene encoding           mitochondrial protein, partial cds       U37101_at   U37101 RRU37101  Rattus rattus     436   403   59   179   2.1           granulocyte colony stimulating factor           mRNA, complete cds       U50185_g_at   U50185 RNU50185  Rattus norvegicus     345   446   229   197   1.8           kidney protein phosphatase 1 myosin           binding subunit mRNA, partial cds       U84402_at   U84402 RNU84402  Rattus norvegicus     537   611   256   219   2.4           smoothened mRNA, complete cds       U92284_at   U92284  Rattus norvegicus  GABA-A   216   210   78   72   −2.9           receptor epsilon subunit gene, partial           cds/cds = 0.1154/gb = U92284/           gi = 2735328/ug = Rn.10869/len = 1600       X14848cds#12_at   X14848cds#12 MIRNXX  Rattus     461   379   218   221   1.9             norvegicus  mitochondrial genome       X56325mRNA_s_at   X56325mRNA RN2A1GL  R. norvegicus     2029   1486   1069   976   1.7           2-alpha-1 globin gene       X58294_at   X58294  R. norvegicus  mRNA for   88   254   387   426   −1.8           carbonic anhydrase II/cds = 8.790/           gb = X58294/gi = 55837/ug = Rn.3525/           len = 1459       X62086mRNA_s_at   X62086 mRNA RNCYP3A1   236   254   433   599   −2.1             R. norvegicus  CYP3A1 gene for           cytochrome P450 PCN1       X69903_at   X69903  R. norvegicus  mRNA for   417   408   146   118   −3.1           interleukin 4 receptor/cds = 9.2411/           gb = X69903/gi = 56390/ug = Rn.10471/           len = 2450       X89968_g_at   X89968 RNSNAPGEN  Rattus     471   555   929   1080   −2.0             norvegicus  mRNA for alpha-soluble           NSF attachment protein                  
 
         [0198]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and its corresponding control in the medial prefrontal cortex (mPFC). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                           TABLE 12                           mPFC Fold Change 1-Week Withdrawal to Control                    1-week   1-week   1-week   1-week                   withdrawal   withdrawal   withdrawal   withdrawal   Fold       Experiment   Description   control A   control B   A   B   change                    AB006450_at   AB006450  Rattus norvegicus  mRNA for   167   198   409   326   1.8           Tim17, complete cds/cds = 4,519/           gb = AB006450/gi = 2335036/           ug = Rn.2099/len = 944       AB020504_at   AB020504  Rattus norvegicus  mRNA for   181   202   385   361   2.0           PMF31, complete cds       AF001898_at   AF001898  Rattus norvegicus  aldehyde   796   993   518   453   −1.8           dehydrogenase (ALDH) mRNA,           complete cds/cds = 28,1533/           gb = AF001898/gi = 2183216/           ug = Rn.6132/len = 2095       AF091566_f_at   AF091566  Rattus norvegicus  isolate   −151   −48   407   303   1.8           HTF-SP1 olfactory receptor mRNA,           partial cds       D28111_at   D28111 RATMAOBP2 Rat mRNA for   796   860   210   220   3.9           MOBP (myelin-associated           oligodendrocytic basic protein),           complete cds, clone rOP1       D28560_at   D28560 RATNPHIII Rat mRNA for   420   393   181   272   −1.7           phosphodiesterase I       K00512_at   K00512 rat myelin basic protein (mbp)   3097   3177   696   689   4.5           gene mrna/cds = UNKNOWN/gb = K00512/           gi = 205320/ug = Rn.9672/len = 1464       L13202_f_at   L13202 RATHFH2  Rattus norvegicus     84   79   208   198   2.5           HNF-3/fork-head homolog-2 (HFH-2)           mRNA, complete cds       L16532_at   L16532  Rattus norvegicus  (clone   867   945   244   233   −3.8           pCNPII) 2′,3′-cyclic nucleotide 3′-           phosphodiesterase (CNPII) mRNA,           complete cds/cds = 79,1341/           gb = L16532/gi = 294526/           ug = Rn.2592/len = 2301       L19180_at   L19180 Rat receptor-linked protein   331   429   46   −13   −1.9           tyrosine phosphatase (PTP-P1) mRNA,           complete cds/cds = 30,4517/           gb = L19180/gi = 310201/           ug = Rn.17237/len = 5396       M11794cds#2_f_at   M11794cds#2 RATMT12C Rat   543   480   894   888   1.7           metallothionein-2 and metallothionein-1           genes, complete cds       M13100cds#1_g_at   M13100cds#1 RATLIN3A Rat long   723   666   1527   1501   2.2           interspersed repetitive DNA sequence           LINE3 (L1Rn)       M13100cds#1_at   M13100cds#1 RATLIN3A Rat long   1805   1436   2832   2939   1.8           interspersed repetitive DNA sequence           LINE3 (L1Rn)       M13100cds#1_g_at   M13100cds#1 RATLIN3A Rat long   666   723   1501   1527   2.2           interspersed repetitive DNA sequence           LINE3 (L1Rn)       M13100cds#5_s_at   M13100cds#5 RATLIN3A Rat long   511   669   1125   1442   2.2           interspersed repetitive DNA sequence           LINE3 (L1Rn)       M20721_f_at   M20721 RATPRPA Rat proline-rich   129   100   282   281   2.5           protein (PRP-1) mRNA, partial cds       M25888_at   M25888 Rat lipophilin mRNA, 3′ end/   4308   3199   1042   1483   −3.0           cds = 0,520/gb = M25888/gi = 206223/           ug = Rn.4550/len = 2585       M36317_s_at   M36317 RATTRHA Rat thyrotropin-   116   132   298   310   2.5           releasing hormone (TRH) precursor           mRNA, complete cds       M60322_at   M60322 Rat aldose reductase gene,   −111   168   562   464   2.6           complete cds/cds = 38,988/gb = M60322/           gi = 202851/ug = Rn.2917/len = 1339       M80570_at   M80570 Rat dopamine transporter   491   387   155   80   −2.2           mRNA, complete cds/cds = 62,1921/           gb = M80570/gi = 310097/ug = Rn.10093/           len = 3386       Rc_AI639204_at   Rat mixed-tissue library  Rattus     309   311   484   606   1.8             norvegicus  cDNA clone rx03840 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_AI639504_at   Rat mixed-tissue library  Rattus     150   151   297   274   1.9             norvegicus  cDNA clone rx04791 3′,           mRNA sequence [ Rattus norvegicus ]       Rc_AA799448_g_at   rc_AA799448 EST188945  Rattus     410   386   197   171   2.2             norvegicus  cDNA, 3′ end/           clone = RHEAB18/clone_end = 3′/           gb = AA799448/gi = 2862403/           ug = Rn.8296/len = 615       Rc_AA800604_g_at   rc_AA800604 EST190101  Rattus     119   232   396   413   1.9             norvegicus  cDNA, 3′ end/clone = RLUAB65/           clone_end = 3′/gb = AA800604/           gi = 2863559/ug = Rn.8590/           len = 579       Rc_AA800693_g_at   rc_AA800693 EST190190  Rattus     749   985   553   441   −1.7             norvegicus  cDNA, 3′ end/           clone = RLUAK36/clone_end = 3′/           gb = AA800693/gi = 2863648/           ug = Rn.6620/ len = 533       Rc_AA818072_s_at   rc_AA818072 UI-R-A0-ag-b-06-0-UI.s2   440   453   178   228   −2.1             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-A0-ag-b-06-0-UI/           clone_end = 3′/gb = AA818072/           gi = 2887952/ug = Rn.11722/len = 408       Rc_AA859643_at   rc_AA859643 UI-R-E0-bs-a-08-0-UI.s1   404   520   193   215   −2.2             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-bs-a-08-0-UI/           clone_end = 3′/gb = AA859643/           gi = 2949163/ug = Rn.32/len = 482       Rc_AA859922_at   rc_AA859922 UI-R-E0-cg-c-04-0-UI.s1   344   413   712   615   1.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cg-c-04-0-UI/           clone_end = 3′/gb = AA859922/           gi = 2949442/ug = Rn.819/len = 373       Rc_AA866432_at   rc_AA866432 UI-R-E0-ch-e-06-0-UI.s1   628   537   302   251   −2.1             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-ch-e-06-0-UI/           clone_end = 3′/gb = AA866432/           gi = 2961893/ug = Rn.3106/len = 484       Rc_AA875411_s_at   rc_AA875411 UI-R-E0-cs-b-11-0-UI.s1   520   476   191   115   −2.5             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cs-b-11-0-UI/           clone_end = 3′/gb = AA875411/           gi = 2980359/ug = Rn.2911/len = 423       Rc_AA875414_at   rc_AA875414 UI-R-E0-cs-d-07-0-UI.s1   218   193   549   634   2.8             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cs-d-07-0-UI/           clone_end = 3′/gb = AA875414/           gi = 2980362/ug = Rn.2912/len = 428       Rc_AA891940_at   rc_AA891940 EST195743  Rattus     427   372   72   143   −2.0             norvegicus  cDNA, 3′ end/           clone = RKIAI82/clone_end = 3′/           gb = AA891940/gi = 3018819/           ug = Rn.3508/len = 523       Rc_AA892942_at   rc_AA892942 EST196745  Rattus     208   192   85   93   2.3             norvegicus  cDNA, 3′ end/           clone = RKIBA19/clone_end = 3′/           gb = AA892942/gi = 3019821/           ug = Rn.3611/len = 511       Rc_AA893593_g_at   rc_AA893593 EST197396  Rattus     357   433   59   −12   −2.0             norvegicus  cDNA, 3′ end/           clone = RPLAC35/clone_end = 3′/           gb = AA893593/gi = 3020472/           ug = Rn.2272/len = 443       Rc_AA945589_at   rc_AA945589 EST201088  Rattus     362   399   860   847   2.2             norvegicus  cDNA, 3′ end/           clone = RLIAP44/clone_end = 3′/           gb = AA945589/ug = Rn.2151/           len = 569       Rc_AA946313_s_at   rc_AA946313 EST201812  Rattus     814   939   445   586   −1.7             norvegicus  cDNA, 3′ end/           clone = RLUBD62/clone_end = 3′/           gb = AA946313/ug = Rn.4295/           len = 505       Rc_AI070277_s_at   rc_AI070277 UI-R-Y0-Is-h-11-0-UI.s1   2415   2557   1169   1337   2.0             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-Y0-Is-h-11-0-UI/           clone_end = 3′/gb = AI070277/           ug = Rn.4550/len = 355       Rc_AI072770_s_at   rc_AI072770 UI-R-Y0-md-g-02-0-UI.s1   1628   1426   327   343   4.6             Rattus norvegicus  cDNA, 3′ end/           clone = UI-R-Y0-md-g-02-0-UI/           clone_end = 3′/gb = AI072770/           ug = Rn.4550/len = 333       Rc_H31839_at   rc_H31839 EST106322  Rattus     292   337   618   681   2.1             norvegicus  cDNA, 3′ end/           clone = RPCAZ43/clone_end = 3′/           gb = H31839/gi = 977256/           ug = Rn.14598/len = 408       U18419_at   U18419  Rattus norvegicus  nonmuscle   209   110   372   357   1.8           caldesmon mRNA, complete cds/           cds = 723,2318/gb = U18419/           gi = 622966/ug = Rn.10621/           len = 5541       U31367_at   U31367  Rattus norvegicus  myelin   488   430   192   238   −2.1           protein MVP17 mRNA, complete cds/           cds = 75,536/gb = U31367/           gi = 914967/ug = Rn.10174/len = 2268       U31866_g_at   U31866  Rattus norvegicus  Nclone10   454   362   111   125   −2.0           mRNA/cds = UNKNOWN/gb = U31866/           gi = 1216376/ug = Rn.11164/len = 2657       U36482_g_at   U36482  Rattus norvegicus     297   400   126   192   −1.7           endoplasmic reticulum protein ERp29           precursor, mRNA, complete cds/           cds = 43,825/gb = U36482/gi = 2317799/           ug = Rn.11262/ len = 1115       U37101_at   U37101 RRU37101 Rattus rattus   167   85   403   436   2.1           granulocyte colony stimulating factor           mRNA, complete cds       U50185_g_at   U50185 RNU50185  Rattus norvegicus     249   182   446   345   1.8           kidney protein phosphatase 1 myosin           binding subunit mRNA, partial cds       U89514_at   U89514  Rattus norvegicus  calpain   219   173   484   350   2.0           large subunit (nCL-4) mRNA, partial           cds/cds = 0,2024/gb = U89514/           gi = 2358261/ug = Rn.10804/len = 2195       X05472cds#1_s_at   X05472cds#1 RNREP24R Rat 2.4 kb   1168   971   1824   2461   2.0           repeat DNA right terminal region       X58294_at   X58294  R. norvegicus  mRNA for   626   592   254   88   −2.7           carbonic anhydrase II/cds = 8,790/           gb = X58294/gi = 55837/ug = Rn.3525/           len = 1459       X61295cds_s_at   X61295cds RNL1RTO2B  R. norvegicus     1759   2259   3785   4678   2.1           L1 retroposon, ORF2 mRNA (partial)       X62086mRNA_s_at   X62086mRNA RNCYP3A1   562   538   254   236   −2.2             R. norvegicus  CYP3A1 gene for           cytochrome P450 PCN1       X69903_at   X69903  R. norvegicus  mRNA for   255   162   408   417   1.8           interleukin 4 receptor/cds = 9,2411/           gb = X69903/gi = 56390/ug = Rn.10471/           len = 2450       X89968_g_at   X89968 RNSNAPGEN  Rattus     928   1171   555   471   −2.0             norvegicus  mRNA for alpha-soluble           NSF attachment protein       Y12502cds_at   Y12502cds RNFXIIIA  R. norvegicus     242   225   85   66   3.1           mRNA for factor XIIIa       Y13381cds_at   Y13381cds RNAMPH1  Rattus     93   73   266   240   3.1             norvegicus  mRNA for amphiphysin,           amph1                  
 
         [0199]     Genes that passed the filtering criteria outlined above for differential expression between 1 week withdrawal and its corresponding control in the medial prefrontal cortex (mPFC). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 13                           VTA 1-Week Extinction to Control                    1-week   1-week   1-week   1-week                               extinction   extinction   extinction   extinction   Mean   Mean       Fold       Probe set no.   Description   control A   control B   B   A   control   exp   Ratio   change                    AF078779_g_at   AF078779  Rattus     652   476   277   381   652   376.5   0.577454   −1.7             norvegicus  putative four           repeat ion channel mRNA,           complete cds       D17614_at   D17614 Rat mRNA for 14-   1249   956   407   344   1249   681.5   0.545637   −1.8           3-3 protein theta-subtype,           complete cds/cds = 85,822/           gb = D17614/gi = 402508/           ug = Rn.2502/len = 2099       rc_AA799299_at   rc_AA799299 EST188796   70   274   542   430   70   408   5.828571   5.8             Rattus norvegicus  cDNA, 3′           end/clone = RHEAA18/           clone_end = 3′/gb = AA799299/           gi = 2862254/ug = Rn.8563/           len = 506       rc_AA893191_at   rc_AA893191 EST196994   55   62   292   313   55   177   3.218182   3.2             Rattus norvegicus  cDNA, 3′           end/clone = RKIBD35/           clone_end = 3′/gb = AA893191/           gi = 3020070/ug = Rn.3301/           len = 654       rc_AA893327_s_at   rc_AA893327 EST197130   58   164   354   429   58   259   4.465517   4.5             Rattus norvegicus  cDNA, 3′           end/clone = RKIBF13/           clone_end = 3′/gb = AA893327/           gi = 3020206/ug = Rn.2732/           len = 452       rc_AA893870_at   rc_AA893870 EST197673   1935   2636   3948   4037   1935   3292   1.701292   1.7             Rattus norvegicus  cDNA, 3′           end/clone = RPLAM86/           clone_end = 3′/gb = AA893870/           gi = 3020749/ug = Rn.11229/           len = 417       rc_AA894330_s_at   rc_AA894330 EST198133   657   469   171   236   657   320   0.487062   −2.1             Rattus norvegicus  cDNA, 3′           end/clone = RSPAW76/           clone_end = 3′/gb = AA894330/           gi = 3021209/ug = Rn.122/           len = 501       rc_AA944856_at   rc_AA944856 EST200355   489   381   187   206   489   284   0.580777   −1.7             Rattus norvegicus  cDNA, 3′           end/clone = REMAQ02/           clone_end = 3′/gb = AA944856/           gi = 3104772/ug = Rn.4992/           len = 339       rc_AI137583_at   rc_AI137583 UI-R-C0-hf-a-   603   482   246   226   603   364   0.603648   −1.7           03-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C0-hf-a-03-0-UI/           clone_end = 3′/gb = AI137583/           ug = Rn.3272/len = 496       rc_H31887_at   rc_H31887 EST106421   573   816   1374   1058   573   1095   1.910995   1.9             Rattus norvegicus  cDNA, 3′           end/clone = RPCBC38/           clone_end = 3′/gb = H31887/           gi = 977304/ug = Rn.14601/           len = 445       S79214cds_s_at   S79214cds type X collagen   457   395   136   267   457   265.5   0.580963   −1.7           alpha 1 chain {NC1 domain}           [rats, Genomic, 491 nt]       S81924_s_at   S81924 Otx1 = homeobox   207   211   −7   27   207   102   0.492754   −2.0           [rats, telencephalon, mRNA           Partial, 444 nt]       U14398_g_at   U14398  Rattus norvegicus     518   483   93   171   518   288   0.555985   −1.8           synaptotagmin IV homolog           mRNA, complete cds/           cds = 267,1544/gb = U14398/           gi = 550453/ug = Rn.11072/           len = 2060       U50842_at   U50842 RNU50842  Rattus     415   376   102   177   415   239   0.575904   −1.7             norvegicus  ubiquitin ligase           (Nedd4) protein mRNA,           partial cds       U52663mRNA#3_s_at   U52663mRNA#3   475   361   181   202   475   271   0.570526   −1.8           RATPAM27  Rattus               norvegicus  peptidylglycine           alpha-amidating           monooxygenase (PAM)           gene, exon 26       X57764_s_at   X57764 Rat mRNA for ET-B   519   427   173   228   519   300   0.578035   −1.7           endothelin receptor/           cds = 203,1528/gb = X57764/           gi = 56122/ug = Rn.11412/           len = 1892       X61106cds_at   X61106cds RNORFEP   229   207   −257   −304   229   −25   −0.10917   9.2             R. norvegicus  ORF for P-           glycoprotein (3′-most exon)           containing epitope for           P-glycoprotein monoclonal           antibody, C219       X96437mRNA_at   X96437mRNA RNPRG1   518   481   99   271   518   290   0.559846   −1.8             R. norvegicus  PRG1 gene                  
 
         [0200]     Genes that passed the filtering criteria outlined above for differential expression between 1 week extinction and its corresponding control in the ventral tegmental area (VTA). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 14                           VTA 1-Week Extinction to Withdrawal                    1-week   1-week   1-week   1-week                               with-   with-   extinction   extinction   Mean   Mean       Fold       Probe set no.   Description   drawal A   drawal B   B   A   control   exp   Ratio   change                    AF037072_at   AF037072  Rattus     770   827   298   360   798.5   329   0.412023   −2.4             norvegicus  carbonic           anhydrase III (CA3) mRNA,           complete cds/cds = 33,815/           gb = AF037072/gi = 2708635/           ug = Rn.22519/len = 1053       D86711_at   D86711 D86711  Rattus     243   252   145   139   247.5   142   0.573737   −1.7             norvegicus  cDNA/gb = D86711/           gi = 1549215/ug = Rn.4240/           len = 994       D88034_at   D88034  Rattus norvegicus     61   61   264   303   61   283.5   4.647541   4.6           mRNA for peptidylarginine           deiminase type III, complete           cds/cds = 42,2036/           gb = D88034/gi = 1644244/           ug = Rn.10658/len = 3100       E02315cds_f_at   E02315cds DNA encoding   2260   2240   799   1028   2250   913.5   0.406   −2.5           calmodulin       L14323_at   L14323  Rattus norvegicus     467   367   109   190   417   149.5   0.358513   −2.8           phospholipase C-beta1b           mRNA, complete alleles/           cds = UNKNOWN/gb = L14323/           gi = 294611/ug = Rn.9741/           len = 7203       Rc_AI639465_f_at   Rat mixed-tissue library   1172   999   466   341   1085.5   403.5   0.371718   −2.7             Rattus norvegicus  cDNA           clone rx01612 3′, mRNA           sequence [Rattus           norvegicus]       Rc_AI639392_at   Rat mixed-tissue library   264   247   84   78   255.5   81   0.317025   −3.2             Rattus norvegicus  cDNA           clone rx02714 3′, mRNA           sequence [ Rattus               norvegicus ]       Rc_AA799410_g_at   rc_AA799410 EST188907   −118   −60   230   232   −89   231   −2.59551   at least             Rattus norvegicus  cDNA, 3′                               2 fold           end/clone = RHEAA81/           clone_end = 3′/gb = AA799410/           gi = 2862365/ug = Rn.3326/           len = 612       Rc_AA894330_s_at   rc_AA894330 EST198133   628   479   171   236   553.5   203.5   0.36766   −2.7             Rattus norvegicus  cDNA, 3′           end/clone = RSPAW76/           clone_end = 3′/gb = AA894330/           gi = 3021209/ug = Rn.122/           len = 501       Rc_AA894345_at   rc_AA894345 EST198148   1220   1203   2191   2183   1211.5   2187   1.8052   1.8             Rattus norvegicus  cDNA, 3′           end/clone = RSPAZ21/           clone_end = 3′/           gb = AA894345/gi = 3021224/           ug = Rn.13530/len = 510       Rc_AA899253_at   rc_AA899253 UI-R-E0-cz-g-   1463   1238   649   682   1350.5   665.5   0.49278   −2.0           07-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E0-cz-g-07-0-           UI/clone_end = 3′/           gb = AA899253/gi = 3034607/           ug = Rn.9560/len = 410       Rc_AI010083_at   rc_AI010083 EST204534   1015   1008   614   528   1011.5   571   0.564508   −1.8             Rattus norvegicus  cDNA, 3′           end/clone = RLUBT52/           clone_end = 3′/gb = AI010083/           ug = Rn.2845/len = 557       Rc_AI137043_at   rc_AI137043 UI-R-C2p-oj-c-   204   226   72   77   215   74.5   0.346512   −2.9           01-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C2p-oj-c-01-0-UI/           clone_end = 3′/gb = AI137043/           ug = Rn.22168/len = 436       Rc_AI137583_at   rc_AI137583 UI-R-C0-hf-a-   613   504   246   226   558.5   236   0.42256   −2.4           03-0-UI.s1  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-C0-hf-a-03-0-U1/           clone_end = 3′/gb = AI137583/           ug = Rn.3272/len = 496       Rc_AI237592_at   rc_AI237592 EST234154   275   263   111   136   269   123.5   0.459108   −2.2             Rattus norvegicus  cDNA, 3′           end/clone = RPLDB22/           clone_end = 3′/gb = AI237592/           ug = Rn.3747/len = 592       S69316_s_at   S69316 S69315S2   764   625   243   242   694.5   242.5   0.349172   −2.9           GRP94/endoplasmin {5′ and           3′ regions} [rats, KNRK           cells, mRNA Partial, 195 nt,           segment 2 of 2]       AFFX_ratb2/X14115_at   X14115 Rat DNA for B2   212   231   43   56   221.5   49.5   0.223476   −4.5           repeat (1-12) from gamma           crystallin gene cluster.       X55298_at   X55298 Rat ribophorin II   164   189   394   396   176.5   395   2.23796   2.2           mRNA/cds = UNKNOWN/           gb = X55298/gi = 57672/           ug = Rn.6863/len = 2234       X61296cds#2_f_at   X61296cds#2 RNL1RTO2C   543   574   63   263   558.5   163   0.291853   −3.4             R. norvegicus  L1 retroposon,           ORF2 mRNA (partial)       X96437mRNA_at   X96437mRNA RNPRG1   487   457   99   271   472   185   0.391949   −2.6             R. norvegicus  PRG1 gene       Z21935cds_at   Z21935cds RNPROKINA   359   332   159   176   345.5   167.5   0.484805   −2.1             R. norvegicus  protein kinase           rMNK2                  
 
         [0201]     Genes that passed the filtering criteria outlined above for differential expression between 1 week withdrawal and 1 week extinction in the ventral tegmental area (VTA). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                                                                                                   TABLE 15                           VTA 1-Week Withdrawal to Control                    R3KJF020   R3KJF020   R3KJF020   R3KJF020   Mean   Mean       Fold       Probe set no.   Description   12264VT   12263VT   12261VT   12262VT   control   exp   Ratio   change                    AA799389_g_at   AA799389 EST188886   398   327   661   694   362.5   677.5   1.868966   1.9             Rattus norvegicus  cDNA, 5′           end/clone = RHEAA70/           clone_end = 5′/gb = AA799389/           gi = 2862344/ug = Rn.3788/           len = 588       AF015305_at   AF015305  Rattus     251   358   590   663   304.5   626.5   2.057471   2.1             norvegicus  equilbrative           nitrobenzylthioinosine-           insensitive nucleoslde           transporter mRNA,           complete cds/cds = 157,1527/           gb = AF015305/gi = 2656138/           ug = Rn.7203/len = 1678       AF064868_g_at   AF064868  Rattus     −125   −223   465   487   −174   476   −2.73563   at least             norvegicus  brain-enriched                               2 fold           guanylate kinase-           associated protein 1 mRNA,           complete cds       AF079162_at   AF079162  Rattus     124   102   369   471   113   420   3.716814   3.7             norvegicus  patched (ptc)           mRNA, partial cds       D84667_at   D84667  Rattus norvegicus     355   430   260   113   392.5   186.5   0.475159   −2.1           mRNA for phosphatidy-           linositol 4-kinase, complete           cds       J03179_at   J03179 Rat D-binding   252   261   152   142   256.5   147   0.573099   −1.7           protein mRNA, complete           cds/cds = 367,1344/           gb = J03179/gi = 203942/           ug = Rn.11274/len = 1622       J03886_at   J03886 Rat skeletal muscle   670   891   1565   1194   780.5   1379.5   1.767457   1.8           myosin light chain kinase,           complete cds/cds = 59,1891/           gb = J03886/gi = 205496/           ug = Rn.9685/len = 2799       K00750exon#2-3_at   K00750exon#2-3 RATCYC   903   713   435   418   808   426.5   0.527847   −1.9           Rat (Sprague-Dawley)           cytochrome c nuclear-           encoded mitochondrial           gene and flanks       L07925_g_at   L07925 RATGNDSA  Rattus     224   168   495   413   196   454   2.316327   2.3             rattus  guanine nucleotide           dissociation stimulator for a           ras-related GTPase mRNA,           complete cds       M33962_g_at   M33962 Rat protein-   201   246   420   391   223.5   405.5   1.814318   1.8           tyrosine-phospatase           (PTPase) mRNA, complete           cds/cds = 119,1417/           gb = M33962/gi = 206496/           ug = Rn.11317/len = 4127       M94918mRNA_f_at   M94918mRNA   1873   1372   3852   3114   1622.5   3483   2.146687   2.1           RATBETGLOX Rat beta-           globin gene, exons 1-3       rc_AI639204_at   Rat mixed-tissue library   247   175   453   331   211   392   1.85782   1.9             Rattus norvegicus  cDNA           clone rx03840 3′, mRNA           sequence [ Rattus               norvegicus ]       rc_AA799571_at   rc_AA799571 EST189068   510   400   105   253   455   179   0.393407   −2.5             Rattus norvegicus  cDNA, 3′           end/clone = RHEAC67/           clone_end = 3′/gb = AA799571/           gi = 2862526/ug = Rn.3458/           len = 541       rc_AA892154_g_at   rc_AA892154 EST195957   224   240   75   54   232   64.5   0.278017   −3.6             Rattus norvegicus  cDNA, 3′           end/clone = RKIAN02/           clone_end = 3′/gb = AA892154/           gi = 3019033/ug = Rn.3279/           len = 386       rc_AA956149_at   rc_AA956149 UI-R-E1-fg-b-   239   206   570   752   222.5   661   2.970787   3.0           03-0-UI.s2  Rattus               norvegicus  cDNA, 3′ end/           clone = UI-R-E1-fg-b-03-0-U1/           clone_end = 3′/gb = AA956149/           ug = Rn.8930/len = 471       rc_AI179445_at   rc_AI179445 EST223155   248   230   119   127   239   123   0.514644   −1.9             Rattus norvegicus  cDNA, 3′           end/clone = RSPCH43/           clone_end = 3′/gb = AI179445/           ug = Rn.221/len = 438       S61973_at   S61973 NMDA receptor   2493   1996   1104   1477   2244.5   1290.5   0.574961   −1.7           glutamate-binding subunit           [rats, mRNA, 1742 nt]       S72637_s_at   S72637 tumor-suppressive   279   202   494   433   240.5   463.5   1.927235   1.9           gene [rats, RSV-trans-           formed 3Y1 fibroblast cells,           SR-3Y1, mRNA, 1788 nt]       U21720mRNA_at   U21720mRNA RNU21720   276   369   564   576   322.5   570   1.767442   1.8             Rattus norvegicus  clone           C201 intestinal epithelium           proliferating cell-associated           mRNA sequence       U88036_at   U88036  Rattus norvegicus     443   415   214   278   429   246   0.573427   −1.7           brain digoxin carrier protein           mRNA, complete cds/           cds = 118,2103/gb = U88036/           gi = 2501807/ug = Rn.5641/           len = 3622       X04070_at   X04070 Rat liver mRNA for   296   279   788   683   287.5   735.5   2.558261   2.6           gap junction protein/           cds = 31,882/gb = X04070/           gi = 56205/ug = Rn.10444/           len = 1485       X60351cds_s_at   X60351cds RRLENSABC   295   227   516   417   261   466.5   1.787356   1.8             R. rattus  mRNA for alpha B-           crystallin (ocular lens tissue)       X61106cds_at   X61106cds RNORFEP   −1   6   262   277   2.5   269.5   107.8   at least             R. norvegicus  ORF for P-                               2 fold           glycoprotein (3′-most exon)           containing epitope for           P-glycoprotein monoclonal           antibody, C219       X70667cds_at   X70667cds RRMC3RA   239   295   432   555   267   493.5   1.848315   1.8             R. rattus  mRNA for           melanocortin-3 receptor                  
 
         [0202]     Genes that passed the filtering criteria outlined above for differential expression between 1 week withdrawal and its corresponding control in the ventral tegmental area (VTA). Average difference values (from GeneChip version 3.2) are listed for each gene from all groups. Affymetrix probe set numbers are listed along with the common name of the genes, if known.  
                         TABLE 16                       Genes that are differentially regulated in various brain       regions in response to extinction and withdrawal       Brain Region                                Nac core           Affymetrix Probe Set #   1 week withdrawal to control           Gene Name       X56729mRNA_at   calpastatin           1 week extinction to control           Gene Name       K02248cds_s_at   somatostatin-14       Z11581_at   kainate receptor subunit (ka2)           1 week extinction to withdrawal           Gene Name       M25890_at   melanocortin-3 receptor       M92076_at   somatostatin       X06564_at   metabotropic glutamate receptor 3       AF102855_at   NCAM           synaptic SAPAP-interacting protein       CeA           1 week withdrawal to control           Gene Name       AB016160_g_at   GABAB receptor 1c       D83538_g_at   phosphatidylinositol 4-kinase           1 week extinction to control           Gene Name       AB016161cds_i_at   GABAB receptor 1d       AF042830_at   tyrosine kinase receptor Ret (c-ret)       E13644cds_s_at   Neurodap-1           1 week extinction to withdrawal           Gene Name       M32754cds_s_at   inhibin alpha-subunit       U14192complete_seq_at   vesicular transport factor       VTA           1 week withdrawal to control           Gene Name       D84667_at   phosphatidylinositol 4-kinase       M33962_g_at   protein-tyrosine-phospatase (PTPase)       S61973_at   NMDA receptor glutamate-binding subunit       X70667cds_at   melanocortin-3 receptor           1 week extinction to control           Gene Name       U14398_g_at   synaptotagmin IV homolog           1 week extinction to withdrawal           Gene Name       E02315cds_f_at   calmodulin       Z21935cds_at   protein kinase rMNK2       L14323_at   phospholipase C-beta1b       Frontal Cortex           1 week withdrawal to control           Gene Name       D28560_at   phosphodiesterase I       L19180_at   tyrosine phosphatase (PTP-P1)       M80570_at   dopamine transporter           1 week extinction to control           Gene Name       D30040_at   RAC protein kinase alpha       K01701_at   oxytocin/neurophysin       M32061_at   alpha-2B-adrenergic receptor       U56261_s_at   SNAP-25a           1 week extinction to withdrawal           Gene Name       D13212_s_at   NMDAR2C       K01701_at   oxytocin/neurophysin       U92284_at   GABA-A receptor epsilon       Nac shell       AJ011318.1   GABA-B receptor subunit gb2       X87900.1   Myelin-associated basic protein       L13041.1   Calcitonin receptor       U92535.1   Bos taurus-like neuronal axonal protein       X98051.1   FRA-2       AI009098   Similar to human oxygen regulated           protein       AI014091   Similar to mouse mrg1 protein       U18772   Pentraxin       U03414   Olfactomedin related protein       U19866.1   Arc - growth factor enriched in dendrites       U28938   Protein tyrosine phosphatase       U67863.1   Melanocortin 4 receptor       U69702.1   ALK-7 kinase       U88958.1   Neuritin       X55812.1   CB1 cannabinoid receptor                  
 
       Example 2  
     Analysis of Western Blots  
       [0203]      FIG. 7  demonstrates that protein levels of gb2 are increased in the nucleus accumbens shell of the 1 week extinction group compared to control animals. This result supports the microarray results and gives stronger evidence for the role of this protein in drug-seeking. In contrast CB-1 protein levels are increased in the nucleus accumbens of the 1 week withdrawal group compared to controls ( FIGS. 8-10 ), though the microarray results showed a decrease. Nevertheless, the results suggest an important role for CB-1 in drug-seeking.  
         [0204]     It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference for all purposes.