Peter J. Sheridan

The D2 dopamine receptor gene as a determinant of reward deficiency syndrome.

The dopaminergic system, and in particular the dopamine D2 receptor, has been profoundly implicated in reward mechanisms in the brain. Dysfunction of the D2 dopamine receptors leads to aberrant substance seeking behaviour (alcohol, drug, tobacco, and food) and other related behaviours (pathological gambling, Tourettes syndrome, and attention deficit hyperactivity disorder). We propose that variants of the D2 dopamine receptor gene are important common genetic determinants of the ‘reward deficiency syndrome’.

Association of the A1 allele of the D2 dopamine receptor gene with severe alcoholism.

In a blinded study, 159 subjects composed of nonalcoholics (N = 43), less severe alcoholics (N = 44), severe alcoholics (N = 52) and young children of alcoholics (CoAs, N = 20) were studied for their allelic association with the D2 dopamine receptor (D2DR) gene utilizing peripheral lymphocytes as the DNA source. The combined alcoholic group compared to the nonalcoholic group showed a significantly greater association with the A1 allele of the D2DR gene. Furthermore, an even more robust association was found when severe alcoholics were compared to nonalcoholics. CoAs also showed a significantly greater association with the A1 allele than nonalcoholics but not when compared to alcoholics. Analysis of risk of alcoholism severity suggests that it comprises of two independent components: family history of alcoholism and presence of the A1 allele. Genotype and allelic frequency of the D2DR gene were also analyzed with respect to race. A higher percentage of blacks compared to whites had the A1/A1 genotype, and A1 allelic frequency in the total sample of blacks was significantly greater than in the total sample of whites. Moreover, frequency of the A1 allele was significantly greater in severe alcoholics compared to nonalcoholics in both whites and blacks. However, due to the small sample size of blacks, these racial differences need to be further studied. This study, of the largest sample of alcoholics to date, strongly affirms association of severe alcoholism with the A1 allele of the D2DR gene.

Nuclear uptake of sex steroid hormones in the cardiovascular system of the baboon.

Cardiac and arterial tissues of six male and six female adult baboons were examined for nuclear uptake of tritiated 5-a-dihydrotestosterone (3H-DHT) or tritiated estradiol-17/S (H-E2) by autoradiography.3H-DHT uptake occurred in nuclei of most atrial and ventricular myocardial fibers, no cardiac interstitial tissues, some arterial endothelial cells, most smooth muscle cells of the intima and inner arterial media, and a few smooth muscle cells of the outer arterial media. 3H-E2 uptake occurred in nuclei of a few atrial and ventricular myocardial fibers, many cardiac interstitial cells, occasional arterial endothelial cells, a few smooth muscle cells of the intima and inner arterial media, smooth muscle cells of the outer arterial media, and nearly all adventitial cells. These observations are consistent with other autoradiographic and biochemical findings which indicate that the heart and major arteries of several mammalian species contain androgen and estrogen receptors in distinctive patterns of distribution among muscle and connective tissue cells. Circ Res 48: 238-244, 1981

Increased prevalence of the Taq I A1 allele of the dopamine receptor gene (DRD2) in obesity with comorbid substance use disorder: a preliminary report.

In order to investigate the prevalence of the Taq I A1 allele of the dopamine receptor gene (DRD2) in obesity with and without comorbid substance use disorder, a total of 40 patients, from an outpatient neuropsychiatric clinic in Princeton, New Jersey, were genotyped for presence or absence of the Taq I DRD2 A1 allele. The primary inclusion criterion for 40 obese subjects was a body mass index (BMI) equal to or over 25 (uncharacterized); 11 obese subjects had severe substance use disorder; 20 controls had a BMI below 25; and, 33 substance use disorder (less severe) patients had a BMI below 25. The data were statistically compared with three different sets of controls divided into three separate groups (Group I, n = 20; Group II, n = 286; Group III, n = 714). They differed according to screening criteria (drug, alcohol, nicotine abuse/dependence, BMI below 25 and other related behaviours including parental history of alcoholism or drug abuse and DSM IV, Axis I and Axis II diagnoses). Groups II and III were population controls derived from the literature. The prevalence of the Taq I A1D2 dopamine receptor (DRD2) alleles was determined in 40 Caucasian obese females and males. In this sample with a mean BMI of 32.35 +/- 1.02, the A1 allele of the DRD2 gene was present in 52.5% of these obese subjects. Furthermore, we found that in the 23 obese subjects possessing comorbid substance use disorder, the prevalence of the DRD2 A1 allele significantly increased compared to the 17 obese subjects without comorbid substance use disorder. The DRD2 A1 allele was present in 73.9% of the obese subjects with comorbid substance use disorder compared to 23.5% in obese subjects without comorbid substance use disorder. Moreover, when we assessed severity of substance usage (alcoholism, cocaine dependence, etc.) increasing severity of drug use increased the prevalence of the Taq I DRD2 A1 allele; where 66.67% (8/12) of less severe probands possessed the A1 allele compared to 82% (9/11) of the most severe cases. Linear trend analyses showed that increasing use of drugs was positively and significantly associated with A1 allelic classification (p < 0.00001). These preliminary data suggest that the presence of the DRD2 A1 allele confirms increased risk not only for obesity, but also for other related addictive behaviours (previously referred to as the Reward Deficiency Syndrome) and that a BMI over 25 by itself (without characterization of macroselection or comorbid substance use disorders) is not a sufficient criterion for association with the DRD2 A1 allele.

Loss of heterozygosity on chromosome 10 in human glioblastoma multiforme

Recessive mutations, revealed by loss of the wild-type allele, have been associated with the development of a variety of cancers in children and adults. Polymorphic chromosome 10 markers were used to screen paired tumor and lymphocyte DNA samples in 13 patients with glioblastoma multiforme. Ten patients showed loss of constitutional heterozygosity in the tumor samples. This finding suggests that a recessive gene involved in the development of glioblastoma multiforme is present on chromosome 10.

Sexual dimorphism in the distribution of estrogen receptors in the temporomandibular joint complex of the baboon

The localization of radiolabeled estradiol was examined in the temporomandibular complex of male baboons by means of an autoradiographic technique. Five baboons were studied. Four animals received only the tritiated estrogen (1 microgram/kgm) and one animal, which served as the control, received both the tritiated estrogen and the unlabeled estrogen (100 micrograms/kgm). The study failed to demonstrate nuclear uptake and retention of tritiated estrogen in any of the tissues of the temporomandibular joint complex, including the articular surface of the condyle, articular disk, capsule, and muscles of mastication. However, estrogen receptors were identified in other tissues, including the pituitary. All tissues examined in the control animal were negative for estrogen receptors. It was concluded that there were no estrogen receptors in the temporomandibular joint complex of aged male baboons. As in previous studies, these findings provide additional evidence of a sexual dimorphism with respect to estrogen receptor distribution in the temporomandibular joint complex of the baboon. Furthermore, it is reasonable to speculate that estrogens may modulate a variety of metabolic functions in these tissues that could be important in the maintenance, repair, and/or pathogenesis of the temporomandibular joint.

Estrogen receptors in the temporomandibular joint of the baboon (Papio cynocephalus): An autoradiographic study☆

Using an autoradiographic method, the temporomandibular joint (TMJ) complex of five aged female baboons was studied for the presence of receptors for estradiol-17 beta. The study was performed in an effort to learn more of the pathophysiology of this joint and in an attempt to provide a scientific basis to explain the reported preponderance of women who seek and undergo treatment for signs and symptoms referable to the TMJ. This experiment revealed that the TMJ complex contains numerous cells with receptors for estrogen, particularly the articular surface of the condyle, articular disk, and capsule. Muscles of mastication contained relatively fewer receptors. As a result, one may postulate a role for the sex steroid hormones in the maintenance, repair, and/or pathogenesis of the TMJ. Additional studies are necessary to fully determine the significance of hormone receptors in this site and any correlation between diseases of the TMJ and the endocrine status of affected patients.

Towards a new model for the mechanism of action of steroids

The classical model for the mechanisms of action of steroids holds that unbound receptors for steroids reside exclusively in the cytoplasmic compartment and that they undergo translocation to the nucleus when bound to steroids in a process which is temperature sensitive. In the present study we looked at the localization of the estradiol receptor using autoradiography and biochemical procedures. Uteri from ovariectomized, and/or ovariectomized-adrenalectomized rats, as well as several cell lines [with estrogen receptors (ER+)] were incubated with [3H]-estradiol or [3H]-R2858 (methoxy-17-ethinylestradiol) for 5 min to 2 h at different concentrations of ligands (0.1-10 nM) at 4 degrees C. When the tissue or cells were processed for autoradiography the localization of steroid was nuclear and cytoplasmic. In contrast when the tissue or cells were processed using the usual biochemical procedures, all binding activity appeared in the cytosolic fraction. In addition, when concentrated preparations of homogenized uteri as well as several nuclear preparation from cell lines were made, free receptor could be demonstrated in the crude nuclear preparations. In the present study, data are presented which suggest that there are unbound receptors for estrogen in nuclei of the smooth muscle cells of myometrium as well as in nuclei of the several cell lines (ER+). We propose a new model for the distribution of estrogen receptors in which unbound receptors are in equilibrium, partitioned between nucleus and cytoplasm according to the free water content of these intracellular compartments.

Androgen receptor in the rat brain — assays and properties

The existence and relevance of an androgen receptor in the developing brain has been a matter of controversy. We here describe both sucrose density gradient and hydroxylapatite assays which clearly define a distinct androgen receptor in the 24-day-old rat hypothalamus, amygdala and preoptic region, but not in the cortex. This receptor has considerable affinity for estradiol-17beta, thus perhaps accounting for some uncertainty about its nature, but none for diethylstilbestrol or other estrogens, antiestrogens or glucocorticoids. Its Kd for both dihydrotestosterone and testosterone is about 1 X 10(-9) M and for estradiol about 2 X 10(-8) M. Its properties are generally consistent with those of androgen receptor reported for other tissues.

Sex steroid receptors in the stomach, liver, pancreas, and gastrointestinal tract of the baboon

Sex steroids have been shown to have a marked effect on the physiologic activities of the liver and the gastrointestinal tract. We performed autoradiographic studies using [3H]estradiol and [3H]dihydrotestosterone on male and female baboons for the purpose of identifying estrogen or androgen receptors, or both, in the liver, pancreas, stomach, and small and large intestines of baboons. Evidence for the presence of estrogen and androgen receptors was made apparent by high concentrations of silver grains over the nuclei of the cells of these tissues. Androgen receptors were largely confined to the nuclei of the smooth muscle cells of the tunica muscularis of the gut wall and the connective tissue interstitial cells of the liver, pancreas, stomach, and intestines. Estrogen receptors were prominent in the nuclei of the vascular smooth muscle cells in the liver, pancreas, gut, and the majority of the endocrine islet cells. These observations suggest that a variety of different cell types of the liver, pancreas, and gastrointestinal tract contain estrogen and androgen receptors that might modulate their cellular activities and influence several different physiologic processes.