preproenkephalin and Cocaine-Related-Disorders

Addiction to opiates and illicit use of psychostimulants is a chronic, relapsing brain disease that, if left untreated, can cause major medical, social, and economic problems. This article reviews recent progress in studies of association of gene variants with vulnerability to develop opiate and cocaine addictions, focusing primarily on genes of the opioid and monoaminergic systems. In addition, we provide the first evidence of a cis-acting polymorphism and a functional haplotype in the PDYN gene, of significantly higher DNA methylation rate of the OPRM1 gene in the lymphocytes of heroin addicts, and significant differences in genotype frequencies of three single-nucleotide polymorphisms of the P-glycoprotein gene (ABCB1) between "higher" and "lower" methadone doses in methadone-maintained patients. In genomewide and multigene association studies, we found association of several new genes and new variants of known genes with heroin addiction. Finally, we describe the development and application of a novel technique: molecular haplotyping for studies in genetics of drug addiction.

Topics: ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; Catechol O-Methyltransferase; Cocaine-Related Disorders; Enkephalins; Epigenesis, Genetic; Genetic Markers; Genetic Variation; Genome-Wide Association Study; Haplotypes; Humans; Methadone; Opioid-Related Disorders; Pharmacogenetics; Protein Precursors; Receptor, Melanocortin, Type 2; Receptor, Serotonin, 5-HT1B; Receptors, Opioid, kappa; Receptors, Opioid, mu; Tryptophan Hydroxylase

The aim of this secondary analysis was to identify prodynorphin (PDYN) genetic markers moderating the therapeutic response to treatment of cocaine dependence with buprenorphine/naloxone (Suboxone®; BUP).. Cocaine-dependent participants (N = 302) were randomly assigned to a platform of injectable, extended-release naltrexone (XR-NTX) and one of three daily medication arms: 4 mg BUP (BUP4), 16 mg BUP (BUP16), or placebo (PLB) for 8 weeks (Parent Trial Registration: Protocol ID: NIDA-CTN-0048, Clinical Trials.gov ID: NCT01402492). DNA was obtained from 277 participants. Treatment response was determined from weeks 3 to 7 over each 1-week period by the number of cocaine-positive urines per total possible urines.. In the cross-ancestry group, the PLB group had more cocaine-positive urines than the BUP16 group (P = 0.0021). The interactions of genetic variant × treatment were observed in the rs1022563 A-allele carrier group where the BUP16 group (N = 35) had fewer cocaine-positive urines (P = 0.0006) than did the PLB group (N = 26) and in the rs1997794 A-allele carrier group where the BUP16 group (N = 49) had fewer cocaine-positive urines (P = 0.0003) than did the PLB group (N = 58). No difference was observed in the rs1022563 GG or rs1997794 GG genotype groups between the BUP16 and PLB groups. In the African American-ancestry subgroup, only the rs1022563 A-allele carrier group was associated with treatment response.. These results suggest that PDYN variants may identify patients who are best suited to treatment with XR-NTX plus buprenorphine for cocaine use disorder pharmacotherapy.

Topics: Buprenorphine; Buprenorphine, Naloxone Drug Combination; Cocaine; Cocaine-Related Disorders; Delayed-Action Preparations; Enkephalins; Humans; Injections, Intramuscular; Naltrexone; Narcotic Antagonists; Opioid-Related Disorders; Protein Precursors

Early-life stressful events affect the neurobiological maturation of cerebral circuitries including the endogenous opioid system and the effects elicited by adolescent cocaine exposure on this system have been poorly investigated. Here, we evaluated whether cocaine exposure during adolescence causes short- or long-term alterations in mRNAs codifying for selected elements belonging to the opioid system. Moreover, since brain-derived neurotrophic factor (BDNF) may undergo simultaneous alterations with the opioid peptide dynorphin, we also evaluated its signaling pathway as well.. Adolescent male rats were exposed to cocaine (20 mg/kg/day) from post-natal day (PND) 28 to PND42, approximately corresponding to human adolescence. After short- (PND45) or long-term (PND90) abstinence, prodynorphin-κ-opioid receptor (pDYN-KOP) and pronociceptin-nociceptin receptor (pN/OFQ-NOP) gene expression were evaluated in the nucleus accumbens (NAc) and hippocampus (Hip) together with the analysis of BDNF signaling pathways.. In the NAc of PND45 rats, pDYN mRNA levels were up-regulated, an effect paralled by increased BDNF signaling. Differently from NAc, pDYN mRNA levels were down-regulated in the Hip of PND45 rats without significant changes of BDNF pathway. At variance from PND45 rats, we did not find any significant alteration of the investigated parameters either in NAc and Hip of PND90 rats.. Our results indicate that the short-term withdrawal from adolescent cocaine exposure is characterized by a parallel pDYN mRNA and BDNF signaling increase in the NAc. Given the depressive-like state experienced during short abstinence in humans, we hypothesize that such changes may contribute to promote the risk of cocaine abuse escalation and relapse.

Topics: Animals; Brain-Derived Neurotrophic Factor; Cocaine-Related Disorders; Dynorphins; Enkephalins; Gene Expression; Male; Nociceptin Receptor; Nucleus Accumbens; Opioid Peptides; Protein Precursors; Rats; Rats, Sprague-Dawley; Receptors, Opioid; Receptors, Opioid, kappa; RNA, Messenger; Signal Transduction; Substance Withdrawal Syndrome

Genetic factors are believed to account for 30-50% of the risk for cocaine and heroin addiction. Dynorphin peptides, derived from the prodynorphin (PDYN) precursor, bind to opioid receptors, preferentially the kappa-opioid receptor, and may mediate the aversive effects of drugs of abuse. Dynorphin peptides produce place aversion in animals and produce dysphoria in humans. Cocaine and heroin have both been shown to increase expression of PDYN in brain regions relevant for drug reward and use. Polymorphisms in PDYN are therefore hypothesized to increase risk for addiction to drugs of abuse. In this study, 3 polymorphisms in PDYN (rs1022563, rs910080 and rs1997794) were genotyped in opioid-addicted [248 African Americans (AAs) and 1040 European Americans (EAs)], cocaine-addicted (1248 AAs and 336 EAs) and control individuals (674 AAs and 656 EAs). Sex-specific analyses were also performed as a previous study identified PDYN polymorphisms to be more significantly associated with female opioid addicts. We found rs1022563 to be significantly associated with opioid addiction in EAs [P = 0.03, odds ratio (OR) = 1.31; false discovery rate (FDR) corrected q-value]; however, when we performed female-specific association analyses, the OR increased from 1.31 to 1.51. Increased ORs were observed for rs910080 and rs199774 in female opioid addicts also in EAs. No statistically significant associations were observed with cocaine or opioid addiction in AAs. These data show that polymorphisms in PDYN are associated with opioid addiction in EAs and provide further evidence that these risk variants may be more relevant in females.

Topics: Adolescent; Adult; Alleles; Behavior, Addictive; Black or African American; Cocaine-Related Disorders; Enkephalins; Female; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Heroin Dependence; Humans; Linkage Disequilibrium; Male; Middle Aged; Polymorphism, Single Nucleotide; Protein Precursors; Sex Factors; White People

A polymorphic 68-bp tandem repeat has been identified within the promoter of the human prodynorphin (PDYN) gene. We found that this 68-bp repeat in the PDYN promoter occurs naturally up to five times. We studied the effect of the number of 68-bp repeats, and of a SNP (rs61761346) found within the repeat on PDYN gene promoter activity. Thirteen promoter forms, different naturally occurring combinations of repeats and the internal SNP, were cloned upstream of the luciferase reporter gene, transfected into human SK-N-SH, H69, or HEK293 cells. Cells were then stimulated with TPA or caffeine. We found cell-specific effects of the number of 68-bp repeats on the transcriptional activity of the PDYN promoter. In SK-N-SH and H69 cells, three or four repeats led to lower expression of luciferase than did one or two repeats. The opposite effect was found in HEK293 cells. The SNP also had an effect on PDYN gene expression in both SK-N-SH and H69 cells; promoter forms with the A allele had significantly higher expression than promoter forms with the G allele. These results further our understanding of the complex transcriptional regulation of the PDYN gene promoter.

Topics: Alcoholism; Alleles; Base Pairing; Cell Line; Cloning, Molecular; Cocaine-Related Disorders; Enkephalins; Genetic Variation; Genotype; HEK293 Cells; Humans; Polymorphism, Genetic; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Protein Precursors; Tandem Repeat Sequences; Transcription, Genetic; Transfection

Dynorphin peptides and the kappa-opioid receptor are important in the rewarding properties of cocaine, heroin, and alcohol. We tested polymorphisms of the prodynorphin gene (PDYN) for association with cocaine dependence and cocaine/alcohol codependence. We genotyped six single nucleotide polymorphisms (SNPs), located in the promoter region, exon 4 coding, and 3' untranslated region, in 106 Caucasians and 204 African Americans who were cocaine dependent, cocaine/alcohol codependent, or controls. In Caucasians, we found point-wise significant associations of 3'UTR SNPs (rs910080, rs910079, and rs2235749) with cocaine dependence and cocaine/alcohol codependence. These SNPs are in high linkage disequilibrium, comprising a haplotype block. The haplotype CCT was significantly experiment-wise associated with cocaine dependence and with combined cocaine dependence and cocaine/alcohol codependence (false discovery rate, q=0.04 and 0.03, respectively). We investigated allele-specific gene expression of PDYN, using SNP rs910079 as a reporter, in postmortem human brains from eight heterozygous subjects, using SNaPshot assay. There was significantly lower expression for C allele (rs910079), with ratios ranging from 0.48 to 0.78, indicating lower expression of the CCT haplotype of PDYN in both the caudate and nucleus accumbens. Analysis of total PDYN expression in 43 postmortem brains also showed significantly lower levels of preprodynorphin mRNA in subjects having the risk CCT haplotype. This study provides evidence that a 3'UTR PDYN haplotype, implicated in vulnerability to develop cocaine addiction and/or cocaine/alcohol codependence, is related to lower mRNA expression of the PDYN gene in human dorsal and ventral striatum.

Topics: Alcohol-Related Disorders; Black or African American; Brain; Cocaine-Related Disorders; Enkephalins; Gene Expression; Gene Frequency; Genetic Predisposition to Disease; Haplotypes; Humans; Polymorphism, Single Nucleotide; Protein Precursors; RNA, Messenger; White People

Prior activation of the kappa opioid system by repeated stress or agonist administration has been previously shown to potentiate the rewarding properties of subsequently administered cocaine. In the present study, intermittent and uncontrollable footshock, a single session of forced swim, or acute administration of the kappa agonist U50,488 (5 mg/kg) were found to reinstate place preference in mice previously conditioned with cocaine (15 mg/kg) and subsequently extinguished by repeated training sessions without drug.. Stress-induced reinstatement did not occur for mice pretreated with the kappa opioid receptor antagonist norbinaltorphimine (10 mg/kg) and did not occur in mice lacking either kappa opioid receptors (KOR -/-) or prodynorphin (Dyn -/-). In contrast, the initial cocaine conditioning and extinction rates were not significantly affected by disruption of the kappa opioid system. Cocaine-injection also reinstated conditioned place preference in extinguished mice; however, cocaine-primed reinstatement was not blocked by kappa opioid system disruption.. The results suggest that stress-induced drug craving in mice may require activation of the dynorphin/kappa opioid system.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Behavior, Animal; Cocaine-Related Disorders; Conditioning, Operant; Disease Models, Animal; Enkephalins; Extinction, Psychological; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Protein Precursors; Receptors, Opioid, kappa; Stress, Psychological; Swimming

There is strong evidence for a genetic contribution to individual differences in vulnerability to drug addictions. Studies have shown that the 68-base pair repeat polymorphism in the promoter region of the human prodynorphin gene contains a putative AP-1 binding site, and that three or four repeat copies result in greater transcriptional activation. Here, we report on a separate cohort of 302 subjects ascertained and characterized extensively by Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition and Addiction Severity Index criteria as: (1) a control group of 127 subjects with no history of alcohol or drug abuse or dependence; (3) a case group of 82 with cocaine dependence only; and (3) a case group of 93 with cocaine and alcohol codependence. The promoter region of the prodynorphin gene containing the repeat was amplified from genomic DNA by polymerase chain reaction and analyzed via gel electrophoresis. Statistical tests were performed with data stratified by the three major ethnic groups studied: African American, Caucasian and Hispanic. For analyses, genotypes were grouped into short (1,1; 1,2; 2,2), short/long (1,3; 2,3; 1,4; 2,4) and long (3,3; 3,4; 4,4) repeats. Deviation from Hardy-Weinberg Equilibrium in the African American control group necessitated testing for association using grouped genotypes rather than grouped alleles. In controls, a significant difference was found in grouped genotype distribution among ethnicities. We found a point-wise, but not experiment-wise across-ethnicities, significant difference in grouped genotype frequency between the cocaine/alcohol-codependent group and the controls in African Americans, with genotypes containing longer alleles found at higher frequency in the codependent group.

Topics: Alcoholism; Alleles; Base Pairing; Black People; Cocaine-Related Disorders; Comorbidity; DNA; Enkephalins; Female; Gene Frequency; Genotype; Hispanic or Latino; Humans; Male; Polymerase Chain Reaction; Promoter Regions, Genetic; Protein Precursors; Tandem Repeat Sequences; White People

The Flinders sensitive line (FSL) rat is a proposed genetic hypercholinergic animal model of human depression. Considering the strong comorbidity between depression and cocaine dependence we investigated the well-documented behavioral and molecular effects of cocaine in the FSL and their control Flinders resistant line (FRL) rats. First, we found no difference between the two lines to establish cocaine self-administration; both lines reached stable responding within 10 days of training at a fixed ratio-1 schedule of reinforcement (1.5 mg/kg/injection). However, the FSL rats exhibited reduced cocaine intake at a dose of 0.09 mg/kg/injection in a within-session dose-response curve (0.02, 0.09, 0.38, 1.5 mg/kg/injection). Second, we examined the effects of repeated cocaine administration on locomotor activity, dopamine overflow and striatal prodynorphin mRNA expression. We found the FSL rats to be low responders to novelty and to exhibit less locomotor activation after repeated cocaine administration (30 mg/kg, i.p., daily injections for 10 days) than their controls. Microdialysis sampling from the nucleus accumbens shell revealed no significant difference in the dopamine overflow between the rat lines, neither during baseline nor after cocaine stimulation. Postmortem analyses of striatal prodynorphin mRNA expression (using in situ hybridization histochemistry) revealed a differentiated response to the cocaine exposure. In contrast to control FRL rats, the FSL rats showed no typical cocaine-evoked elevation of prodynorphin mRNA levels in rostral subregions of the striatum whereas both strains expressed increased prodynorphin mRNA levels in the caudal striatum after cocaine administration. In conclusion, the FSL animal model of depression demonstrates marked blunting of the locomotor and dynorphin neuroadaptative responses to cocaine in accordance with its enhanced cholinergic sensitivity.

Topics: Acetylcholine; Animals; Chromatography, High Pressure Liquid; Cocaine; Cocaine-Related Disorders; Corpus Striatum; Depression; Disease Models, Animal; Dopamine; Dopamine Uptake Inhibitors; Enkephalins; In Situ Hybridization; Male; Microdialysis; Motor Activity; Nucleus Accumbens; Protein Precursors; Rats; RNA, Messenger

An increase in preprodynorphin (ppdyn) mRNA was detected in the caudate putamen of chronically cocaine-treated and 3-h withdrawn rats. An increase in mu-opioid receptor (MOP) mRNA levels was observed in the frontal cortex of 3-h withdrawn rats. Naloxone had no effect on the increase of MOP or ppdyn mRNA levels. The results indicate that the opioid system is altered during early withdrawal from chronic cocaine administration.

Topics: Animals; Brain Chemistry; Cocaine; Cocaine-Related Disorders; Disease Models, Animal; Dopamine Uptake Inhibitors; Down-Regulation; Drug Administration Schedule; Enkephalins; Frontal Lobe; Male; Narcotic Antagonists; Protein Precursors; Rats; Rats, Inbred F344; Receptors, Opioid, mu; RNA, Messenger; Substance Withdrawal Syndrome; Time Factors

The endogenous opioid system has been shown to have a role in the biological processes involved in addiction to numerous drugs of abuse including cocaine. It has recently been reported that the variable nucleotide tandem repeat (VNTR) polymorphism in the 5' promoter region of the prodynorphin gene, which encodes the precursor for three endogenous opioid peptides, is associated with the cocaine dependent phenotype. In order to confirm this finding, we genotyped the prodynorphin promoter polymorphism in cocaine dependent (n = 167) and control (n = 88) individuals of African descent. The results from this experiment indicate a statistically significant (chi2 = 5.64, OR = 1.59, P = 0.018) association between the prodynorphin promoter VNTR polymorphism and the cocaine dependent phenotype. In contrast to previous work showing increased risk conferred by one or two copies of the prodynorphin VNTR, the genotyping results from this study indicate that persons with three or four copies of this polymorphism are more likely to become cocaine dependent. This disparity suggests that the prodynorphin promoter VNTR may not be the functional polymorphism associating with the cocaine dependent phenotype. It is possible that different alleles of the prodynorphin promoter VNTR in the independent populations used for this and the previous study may be in linkage disequilibrium with a yet to be identified functional polymorphism in this gene.

Topics: Adult; Black or African American; Cocaine-Related Disorders; Enkephalins; Female; Genetic Predisposition to Disease; Humans; Male; Polymorphism, Genetic; Promoter Regions, Genetic; Protein Precursors

The effect of the selective serotonin uptake inhibitor fluoxetine was examined on prodynorphin gene expression. Fluoxetine or vehicle was infused continuously for 7 d via osmotic minipumps into male rats. Northern blot analysis showed significant increases in prodynorphin gene expression in the hypothalamus (171% of controls) and significant decreases in the caudate putamen and nucleus accumbens (62% and 70% of controls, respectively). There were no significant changes in the hippocampus. Thus, chronic inhibition of serotonin uptake can regulate prodynorphin gene expression in the hypothalamus, caudate putamen, and nucleus accumbens. Fluoxetine effects were also evaluated in rats treated with p-chloroamphetamine (PCA), a neurotoxin that depletes serotonin. Because we previously reported that continuous infusion of cocaine for 7 d (which inhibits dopamine, serotonin, and norepinephrine uptake), or GBR 12909 (a selective dopamine uptake inhibitor), produced significant decreases in the hypothalamus and cocaine also produced a significant increase in prodynorphin gene expression in caudate putamen, regulation of prodynorphin gene expression by fluoxetine is suggested to be different from that by cocaine. Because neither a selective dopamine uptake inhibitor nor a selective serotonin uptake inhibitor produced the same effect as cocaine in the caudate putamen, this effect is likely regulated by the inhibition of norepinephrine uptake, by a combination of effects on two or three neurotransmitter transporters, or by a mechanism unrelated to transporter inhibition.

Topics: Animals; Brain; Brain Chemistry; Carrier Proteins; Cocaine; Cocaine-Related Disorders; Dopamine Uptake Inhibitors; Enkephalins; Fluoxetine; Gene Expression Regulation; Male; p-Chloroamphetamine; Protein Precursors; Rats; Rats, Sprague-Dawley; RNA, Messenger; Selective Serotonin Reuptake Inhibitors; Serotonin; Time; Up-Regulation

The investigation of rodent cocaine self-administration (SA) under conditions that promote escalating patterns of intake may provide insight into the loss of control over drug use that is central to human addiction.. This study examines the effects of daily long-access (LgA) SA of high or low cocaine doses on drug intake, extinction, reinstatement, and brain mRNA levels.. Three groups of male Sprague-Dawley rats were trained to self-administer cocaine during multiple-dose sessions. Short-access (ShA) rats were tested daily for multi-dose SA then remained in the chambers for 7 h with no cocaine available. LgA rats had access to low (0.5 mg/kg per infusion; LgA-LD) or high (2.0 mg/kg per infusion; LgA-HD) cocaine doses for 7 h after multi-dose SA. After 14 days, responding was extinguished, cocaine-induced reinstatement was determined, and preproenkephalin (ppENK), preprodynorphin (ppDYN), corticotropin releasing factor (CRF) and dopamine D(2) receptor (D(2)R) mRNA levels were measured in various brain regions using a quantitative solution hybridization RNase protection assay.. Whereas SA was not altered in ShA rats and only increased during the "loading phase" in LgA-LD rats, a general escalation of intake was found in LgA-HD rats. LgA, particularly LgA-HD, rats were more susceptible to reinstatement than ShA rats. Caudate-putamen ppENK and nucleus accumbens D(2)R mRNA levels were elevated in LgA-HD rats. Overall, D(2)R mRNA levels were positively correlated with reinstatement.. The escalation of cocaine SA under LgA conditions is dose-dependent and is associated with heightened susceptibility to drug-induced relapse. The characterization of neurobiological alterations that accompany escalated SA should facilitate the identification of mechanisms underlying the onset of human addiction.

Topics: Animals; Behavior, Addictive; Brain; Cocaine; Cocaine-Related Disorders; Corticotropin-Releasing Hormone; Dose-Response Relationship, Drug; Dynorphins; Enkephalins; Extinction, Psychological; Male; Opioid Peptides; Protein Precursors; Rats; Rats, Sprague-Dawley; Receptors, Dopamine D2; Recurrence; RNA, Messenger; Self Administration; Time Factors

There is evidence that suggests that increased corticotropin-releasing hormone (CRH) release in the central nucleus of the amygdala underlies the anxiogenic and stress-like consequences of withdrawal that are common in phenomenology to all drugs of abuse. The present studies were undertaken to determine levels of CRH mRNA in the amygdala, and also in the hypothalamus, frontal cortex and brainstem after short-term (2 days) and intermediate-term (10 days) cocaine withdrawal (with continued saline injections) from chronic (14 days) 'binge' pattern cocaine administration (3 x 15 mg/kg per day at hourly intervals). Confirming our recent finding of an activation of stress responsive hypothalamic-pituitary-adrenal activity during early cocaine withdrawal, there was a significant elevation of plasma corticosterone level after 2-day cocaine withdrawal. There was also a significant elevation of CRH mRNA levels in the amygdala, but not in the hypothalamus, frontal cortex or brainstem after 2-day cocaine withdrawal. A negative correlation between amygdalar CRH mRNA and plasma corticosterone levels was found in the 2-day cocaine withdrawn rats but not in control rats, suggesting that CRH neurons in the amygdala may be differentially responsive to glucocorticoids after chronic cocaine exposure and withdrawal. There were no changes in either plasma corticosterone or amygdalar CRH mRNA levels after 10-day cocaine withdrawal. Our findings of an increase in amygdalar CRH gene expression during early cocaine withdrawal support a potentially important role for amygdalar CRH activity in the anxiogenic and aversive consequences of withdrawal from cocaine during a time when humans are most subject to relapse.

Topics: Amygdala; Animals; Brain Stem; Chronic Disease; Cocaine; Cocaine-Related Disorders; Corticosterone; Corticotropin-Releasing Hormone; Dopamine Uptake Inhibitors; Enkephalins; Frontal Lobe; Gene Expression; Hypothalamo-Hypophyseal System; Male; Pituitary-Adrenal System; Protein Precursors; Rats; Rats, Inbred F344; RNA, Messenger; Substance Withdrawal Syndrome

Several human and rat studies suggest that the striatal dynorphin system is important for neuroadaptation following cocaine exposure. In the current study, prodynorphin (PDYN) mRNA expression was examined in monkeys at initial and chronic phases of cocaine self-administration. Adult Rhesus monkeys were trained to self-administer food (banana flavoured pellets) or cocaine (0.03 or 0.3 mg/kg per injection) on a fixed interval 3-min schedule for 5 or 100 sessions. Each session ended after 30 reinforcers were delivered. The PDYN mRNA expression was analysed in the precommissural striatum using in situ hybridization histochemistry. We found a specific activation of PDYN mRNA expression in the limbic-innervated patch/striosome compartment of the dorsal caudate and dorsal putamen during the initial (i.e. 5 day) phase of the high dose cocaine self-administration. After 100 days of the high dose exposure, the patch/striosome compartment remained activated, but an increase in PDYN mRNA levels was also evident in the sensorimotor-connected matrix compartment of the caudate. Neither self-administration phase resulted in significant changes in the corresponding striatal regions of the low dose cocaine-exposed primates. Moreover, cocaine self-administration failed to alter the PDYN mRNA expression in high- or low-expressing PDYN cell populations in the nucleus accumbens during any condition studied. These results demonstrate the vulnerability of the dorsal striatum (in particular the caudate) to neuroadaptations following long-term high dose cocaine self-administration. In addition, the temporal nature of the changes in PDYN gene expression within the striatal compartments could reflect a change in drug responsivity that occurs during the transition to drug dependence.

Topics: Animals; Cocaine; Cocaine-Related Disorders; Corpus Striatum; Dopamine Uptake Inhibitors; Enkephalins; Gene Expression; Macaca mulatta; Male; Protein Precursors; RNA, Messenger; Self Administration; Up-Regulation

It has been demonstrated that the opioid peptide dynorphin plays a role in modulating responses to several psychoactive substances including cocaine. Our laboratory and others have found that mRNA levels of dynorphin in the caudate and putamen are elevated after acute or chronic cocaine exposure in rats. Recently, a 68-base pair (bp) repeat polymorphism within the core promoter region of the human prodynorphin gene has been reported to occur in alleles containing one, two, three, or four copies. This repeat contains a putative AP-1 transcription factor binding site; reporter gene constructs with three or four, but not one or two, copies of the tandem repeats were shown to be associated with increases in transcriptional activation in in vitro cellular assays. We hypothesize that this polymorphism may be associated with individual differences in vulnerability to cocaine dependence or abuse. From an ongoing study of the genetics of addiction, 174 subjects were studied, including individuals with a primary diagnosis (DSM-IV criteria) of cocaine dependence (N = 61) or abuse (N = 22), and controls with no history of any substance dependence or abuse (N = 91). We designed primers for polymerase chain reaction (PCR) to amplify sequences of the promoter region of the prodynorphin gene containing the repeat element. The association of alleles containing three or four repeats with cocaine dependence/abuse was examined. With data stratified by ethnic group, pooled relative risk (RR) with Mantel-Haenszel Chi square was calculated: RR = 0.59 (95% confidence interval 0.37-0.95), chi2 (1) = 4.14, P = 0.042. Our results suggest that this allelic variation at the promoter region of the prodynorphin gene (alleles with three or four repeats), which may result in enhanced transcription of the gene, may contribute to relative protection and decrease individual vulnerability to develop cocaine dependence or abuse.

Topics: Alleles; Base Sequence; Cocaine-Related Disorders; DNA; Enkephalins; Gene Frequency; Humans; Molecular Sequence Data; Polymorphism, Genetic; Promoter Regions, Genetic; Protein Precursors; Repetitive Sequences, Nucleic Acid

We previously found that there was a rapid stimulatory effect of acute (1 day) 'binge' cocaine on CRH mRNA levels in the rat hypothalamus. In contrast, after 3 days of 'binge' cocaine, there was a modest decrease (12%) in hypothalamic CRH mRNA levels, which after 14 days of 'binge' cocaine was greater (32%) and significantly lower than control values. Also, our previous studies found an elevation of CRH mRNA in the frontal cortex after 3 days of 'binge' cocaine. The present study was designed to investigate the possible role of dopamine receptors in modulating these effects. Administration of 3 days of 'binge' cocaine (3 x 15 mg/kg, i.p.) was preceded by daily injections of either D(1) (SCH23390, 2 mg/kg) or D(2) (sulpiride, 50 mg/kg) dopamine receptor antagonist. Neither SCH23390 nor sulpiride had an effect on basal CRH mRNA levels in the hypothalamus, frontal cortex or amygdala. Small decreases (10-13%) in hypothalamic CRH mRNA levels were found again to be induced by 3 days of repeated 'binge' cocaine. However, this modest decrease was not found in the rats that received D(1) antagonist SCH23390 pretreatment. Pretreatment with D(2) antagonist sulpiride had no effect on this decrease. These findings suggest that the inhibitory effect of repeated 'binge' cocaine on the hypothalamic CRH mRNA expression is absent when there is D(1), but not D(2), dopamine receptor blockade. In the frontal cortex, pretreatment with either SCH23390 or sulpiride did not alter the increases in the CRH mRNA levels induced by repeated 'binge' cocaine. The results suggest that the cocaine-induced modulation of hypothalamic CRH mRNA expression is secondary to changes in the activity of specific components of dopaminergic systems.

Topics: Amygdala; Animals; Benzazepines; Cocaine; Cocaine-Related Disorders; Corticotropin-Releasing Hormone; Dopamine Antagonists; Dopamine D2 Receptor Antagonists; Dopamine Uptake Inhibitors; Enkephalins; Frontal Lobe; Gene Expression; Hypothalamus; Male; Olfactory Bulb; Protein Precursors; Rats; Rats, Inbred F344; Receptors, Dopamine D1; RNA, Messenger; Sulpiride