Genotyping the -521C/T functional polymorphism in the promoter region of dopamine D4 receptor (DRD4) gene

Rónai, Zsolt; Barta, Csaba; Guttman, András; Lakatos, Krisztina; Gervai, Judit; Staub, Maria; Sasvári-Székely, Mária

doi:10.1002/1522-2683()22:6<1102::aid-elps1102>3.0.co;2-3

Cited by 33 publications

(16 citation statements)

References 26 publications

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“…26,65 After separation by electrophoresis for 3 h on a 2.5% agarose gel, the PCR products (2-8-or 10-repeat units) were sized using a 50 bp ladder. To genotype the DRD4 C-521T polymorphism, PCR was carried out with an allelespecific tetra primer system based on the method of Ronai,66 with slight modifications. Briefly, the reaction mixture contained 1 mM each primer (M3, M4, SN0 and MS0), 66 approximately 30 ng DNA, 200 mM dNTPs, 1.5 mM MgCl 2 and 1 M betaine in a final volume of 25 ml.…”

Section: Genotypingmentioning

confidence: 94%

“…To genotype the DRD4 C-521T polymorphism, PCR was carried out with an allelespecific tetra primer system based on the method of Ronai,66 with slight modifications. Briefly, the reaction mixture contained 1 mM each primer (M3, M4, SN0 and MS0), 66 approximately 30 ng DNA, 200 mM dNTPs, 1.5 mM MgCl 2 and 1 M betaine in a final volume of 25 ml. After an initial denaturation step at 951C for 15 min, thermocycling consisted of 40 cycles of 1 min at 951C, 1 min at 651C and 1 min at 721C, then a final extension step of 721C.…”

Section: Genotypingmentioning

confidence: 99%

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Genetic variation in the dopamine D4 receptor (DRD4) gene and smoking cessation: follow-up of a randomised clinical trial of transdermal nicotine patch

et al. 2007

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Smokers of European ancestry (n ¼ 720) who participated in a double-blind, randomised, placebo-controlled trial of transdermal nicotine replacement therapy, were genotyped for two functional polymorphisms (variable number of tandem repeats (VNTR) and a C to T transition at position -521 (C-521T)) in the dopamine D4 receptor gene (DRD4) gene. Logistic regression models of abstinence at 12-and 26-week follow-ups were carried out separately for each polymorphism. For the DRD4 VNTR models, the main effect of treatment was significant at both 12-week (P ¼ 0.001) and 26-week (P ¼ 0.006) follow-ups, indicating an increased likelihood of successful cessation on active nicotine replacement therapy transdermal patch relative to placebo. The main effect of DRD4 VNTR genotype was associated with abstinence at 12-week follow-up (P ¼ 0.034), with possession of one or more copies of the long allele associated with reduced likelihood of cessation (17 vs 23%), but this effect was not observed at 26-week follow-up. For the DRD4 C-521T models, no main effect or interaction terms involving genotype were retained in the models at either 12-or 26-week follow-up. These data are consistent with observations from studies of the DRD2 gene that genetic variants related to relatively decreased dopaminergic tone in the mesocorticolimbic system are associated with increased risk for relapse to smoking following a cessation attempt.

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Section: Genotypingmentioning

confidence: 94%

Section: Genotypingmentioning

confidence: 99%

Genetic variation in the dopamine D4 receptor (DRD4) gene and smoking cessation: follow-up of a randomised clinical trial of transdermal nicotine patch

et al. 2007

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“…The other technique was a novel, single-step genotyping protocol developed in our laboratory. 32 When the two methods gave different results, the template concentration was varied until results became identical. All the genotyping was done in the Institute of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, blind to the infants' attachment status.…”

Section: Genotypingmentioning

confidence: 99%

Further evidence for the role of the dopamine D4 receptor (DRD4) gene in attachment disorganization: interaction of the exon III 48-bp repeat and the −521 C/T promoter polymorphisms

et al. 2002

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Keywords: DRD4 exon III VNTR polymorphism; −521 C/T promoter polymorphism; attachment disorganization; dopamineIn non-clinical low-risk populations 15% of infants show disorganized attachment behavior 1,2 with their caregivers in the Strange Situation, 3 a mildly stressful laboratory procedure testing infants' ability to cope with separation anxiety. Disorganization of early attachment has been primarily ascribed to inadequate parenting, 2,4,5 and has been associated with childhood behavior problems 6,7 and adolescent psychopathological tendencies. 5We have recently reported an association between the DRD4 exon III 48 basepair repeat polymorphism and disorganization of infants' attachment behavior towards their mother in a low-social-risk group of 1-year-old infants: 8 the risk for disorganized attachment among infants carrying the 7-repeat allele was fourfold. Here we report further evidence for the involvement of the dopamine D4 receptor gene in attachment disorganization. The same group of infants was genotyped for the functional −521 C/T single nucleotide polymorphism (SNP) in the upstream regulatory region of the DRD4 gene 9 in order to test the association with attachment disorganization both alone and in interaction with the DRD4 exon III 7-repeat allele. While the −521 C/T genotype itself had no effect on attachment status ( 2 = 0.41, df = 2, P = 0.82), there was an interaction between the structural 48-bp repeat polymorphism and the −521 C/T promoter polymorphism: the association between disorganized attachment and the 7-repeat allele was enhanced in the presence of the −521 T allele ( 2 = 6.61 and 6.67, df = 1, P Ͻ 0.025 for CT and TT genotypes, respectively). In the presence of both risk alleles the odds ratio for disorganized attachment increased tenfold. This result supports our previous postulation that the DRD4 gene plays a role in the development of attachment behavior in low-risk, non-clinical populations. Molecular Psychiatry (2002) 7, 27-31. DOI: 10.1038/ sj/mp/4000986The role of dopamine receptor and transporter genes in both normal traits and psychopathology has been intensively studied in the past decade.10,11 One of the most frequently targeted candidate genes is the highly polymorphic dopamine D4 receptor (DRD4) gene. 12,13A 48-bp VNTR polymorphism has been identified in exon III with the 4-repeat allele being the most common, followed by the 7-repeat form 14 which is 2-3 times less potent in dopamine-mediated coupling to adenylyl cyclase than the 4-repeat form.15 Evidence linking the 7-repeat allele of the DRD4 gene with infant temperament traits of negative emotionality [16][17][18] and maladaptive behavioral problems, especially attention deficit hyperactivity disorder (ADHD) in children, is being accumulated. 19-21Recently, we found that the 7-repeat DRD4 allele was 2.5 times more frequent among one-year-old infants who showed disorganized attachment behavior, ie were unable to cope with the stress elicited by the two brief separations from the caregiver in the Strange Situation exper...

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“…27 Genotyping Two independent, hot-start PCR methods were used for each genotyping as described elsewhere: The slightly modified PCR-RFLP method described by Okuyama 4 and another novel, single-step genotyping protocol, developed in our laboratory. 5 When the two methods gave different results, repetitions were made by varying the template concentration and/or the enzyme concentration until the results were identical. All the genotyping was done blind to the personality scores.…”

Section: Subjects and Genotypingmentioning

confidence: 99%

“…Moreover, a significantly higher frequency of the CC genotype was found among schizophrenic patients. 15 Previously we have described the distribution of −521 C and −521 T alleles in a nonclinical Hungarian population, 5 which, according to our knowledge, is the first report on a Caucasian population. The obtained allele frequencies (−521 C: 0.43; −521 T: 0.57) were very similar to the Japanese data.…”

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confidence: 99%

Association between Novelty Seeking and the −521 C/T polymorphism in the promoter region of the DRD4 gene

et al. 2000

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Association between the human personality trait 'Novelty Seeking' and the polymorphism of the DRD4 gene was first reported by Ebstein 1 and Benjamin 2 in 1996. This was soon followed by replication studies in various ethnic groups and by studying the role of other neurotransmitter receptor and transporter genes in the genetic determination of human temperament. More recently, several polymorphic sites of the upstream regulatory region of the DRD4 gene have been described.3 Among these the −521 C/T single nucleotide polymorphism (SNP) was shown to be associated with the Novelty Seeking (NS) scores of the Temperament and Character Inventory (TCI) in a Japanese male population. 4 We have investigated the −521 C/T SNP polymorphism in a Caucasian (Hungarian) population, 5 and here we report a replication of the Japanese findings, in an association study involving 109 healthy Hungarian volunteers. We found a weak association between NS and CC vs CT or TT genotypes (P Ͻ 0.06). Examination of this relation in male and female sex groups, however, strengthened the association for females (P Ͻ 0.01), but showed no genotypic effect for males. Molecular Psychiatry (2001) 6, 35-38. Human temperament, as measured by various selfreport questionnaires including TPQ (or a new version known as TCI) 6 and NEO-PI-R, 7 was shown to have considerable inherited components.8 Novelty Seeking (NS) is one of the four dimensions in the psychobiological model of temperament, which are thought to be independently heritable, and manifested from an early age. 9 The NS scale was designed to measure exploratory, impulsive and extravagant behavior, and has been related to the dopamine system by Cloninger. Searching for association between common genetic polymorphisms and specific personality traits, recent studies have concentrated on distinct candidate genes. First, the variable number of 48-bp repeats (VNTR) in the third exon of the human dopamine D4 receptor gene (DRD4) was shown to be correlated with the personality trait of Novelty Seeking, as measured by the TPQ 1 or the NEO-PI-R, 2 although the replications of the initial findings have been controversial. [10][11][12][13] Further studies concerning the regulatory region of the genes encoding receptors and transporters involved in neurotransmission revealed associations between a novel polymorphism of the DRD4 promoter region and NS in Japanese subjects. 4 This polymorphism is a CϾT substitution in a CpG island at one end of a cell-type specific regulatory element, between −591 and −123 bp relative to the first nucleotide of the DRD4 gene.14 Using a transient expression system, the −521 T allele was shown to be approximately 40% less active than the −521 C variant in a human retinoblastoma cell line, 4,15 suggesting the relevance of this polymorphism to the dopaminergic system. Moreover, a significantly higher frequency of the CC genotype was found among schizophrenic patients. 15Previously we have described the distribution of −521 C and −521 T alleles in a nonclinical Hungarian populati...

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Genotyping the -521C/T functional polymorphism in the promoter region of dopamine D4 receptor (DRD4) gene

Cited by 33 publications

References 26 publications

Genetic variation in the dopamine D4 receptor (DRD4) gene and smoking cessation: follow-up of a randomised clinical trial of transdermal nicotine patch

Genetic variation in the dopamine D4 receptor (DRD4) gene and smoking cessation: follow-up of a randomised clinical trial of transdermal nicotine patch

Further evidence for the role of the dopamine D4 receptor (DRD4) gene in attachment disorganization: interaction of the exon III 48-bp repeat and the −521 C/T promoter polymorphisms

Association between Novelty Seeking and the −521 C/T polymorphism in the promoter region of the DRD4 gene

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