Search

Dysfunction of the brain glutamate system has been associated with alcoholism. Ionotropic glutamatergic alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors (AMPARs) play an important role in both neurotransmission and post-synaptic plasticity. Alterations in AMPAR densities may also play a role in the neurobiological changes associated with alcoholism. In the present study, [3H] AMPA binding density was evaluated in the nucleus accumbens (NAc), frontal cortex, anterior cingulate cortex (ACC), dentate gyrus and hippocampus of Cloninger type 1 (n=9) and 2 (n=8) alcoholics, and compared with non-alcoholic control subjects (n=10) by post-mortem whole-hemisphere autoradiography. The [3H] AMPA binding density was significantly higher in the ACC of early onset type 2 alcoholics when compared with controls (p=0.011). There was also a significant negative correlation between [3H] AMPA binding and previously published results of dopamine transporter (DAT) density in the ACC in these same brain samples (R=−0.95, p=0.001). Although preliminary, and from a relatively small diagnostic group, the present results help to further explain the pathology of alcohol dependence and impulsive behaviour in type 2 alcoholics., final draft, peerReviewed

Increased glutamatergic neurotransmission and hyper-excitability during alcoholic withdrawal and abstinence are associated with increased risk for relapse, in addition to compensatory changes in the glutamatergic system during chronic alcohol intake. Type 5 metabotropic glutamate receptor (mGlur5) is abundant in brain regions known to be involved in drug reinforcement, yet very little has been published on mGluR1/5 expression in alcoholics. We evaluated the densities of mGluR1/5 binding in the hippocampus and striatum of post-mortem human brains by using [(3)H]Quisqualic acid as a radioligand in whole hemispheric autoradiography of Cloninger type 1 (n=9) and 2 (n=8) alcoholics and healthy controls (n=10). We observed a 30-40% higher mGluR1/5 binding density in the CA2 area of hippocampus in type 1 alcoholics when compared with either type 2 alcoholics or healthy subjects. Although preliminary, and from a relatively small number of subjects from these diagnostic groups, these results suggest that the mGluR1/5 receptors may be increased in type 1 alcoholics in certain brain areas.

Ethanol modulates the GABA(A) receptor to cause sedative, anxiolytic and hypnotic effects that are qualitatively similar to benzodiazepines and barbiturates. The aim of this study was to explore if GABA(A) receptor density is altered in post-mortem brains of anxiety-prone Cloninger type 1 and socially hostile type 2 alcoholic subtypes when compared to controls. The GABA(A) binding site density was measured by whole-hemisphere autoradiography with tritium labeled flunitrazepam ([(3)H]flunitrazepam) from 17 alcoholic (nine type 1, eight type 2) and 10 non-alcoholic post-mortem brains, using cold flumazepam as a competitive ligand. A total of eight specific brain areas were examined. Alcoholics displayed a significantly (p < 0.001, bootstrap type generalizing estimating equations model) reduced [(3)H]flunitrazepam binding site density when compared to controls. When localized, type 2 alcoholics displayed a significantly (p ≤ 0.05) reduced [(3)H]flunitrazepam binding site density in the internal globus pallidus, the gyrus dentatus and the hippocampus, whereas type 1 alcoholics differed from controls in the internal globus pallidus and the hippocampus. While previous reports have demonstrated significant alterations in dopaminergic and serotonergic receptors between type 1 and type 2 alcoholics among these same subjects, we observed no statistically significant difference in [(3)H]flunitrazepam binding site densities between the Cloninger type 1 and type 2 alcoholics.

The 5-HT(1B) receptor has been associated with alcohol dependence, impulsive or alcohol-related aggressive behavior, and anxiety. The aim of this study was to determine whether or not the 5-HT(1B) receptor density differs in brain samples from anxiety-prone Cloninger type 1 alcoholics and socially hostile, predominantly male, type 2 alcoholics, and controls. Whole-hemispheric 5-HT(1B) receptor density was measured in eight regions of postmortem brains from 17 alcoholics and 10 nonalcoholic controls by autoradiography with tritiated GR-125743 and unlabeled ketanserin to prevent 5-HT(1D) binding. The 5-HT(1B) receptor density was not altered significantly in any of the studied regions. However, some correlations were observed in types 1 and 2 alcoholics only. The 5-HT(1B) receptor density decreased with age in type 1 alcoholics only. There was a significant positive correlation between 5-HT(1B) receptor and serotonin transporter densities in the head of caudate of type 1 alcoholics only. There was a significant positive correlation between 5-HT(1B) receptor density and dopaminergic terminal density, as estimated by vesicular monoamine transporter 2 measurement in the nucleus accumbens of type 2 alcoholics only. There were no significant correlations between 5-HT(1B) receptor and dopamine transporter or dopamine D2/D3 receptor densities in any of the subject groups. In conclusion, these results do not indicate primary changes in 5-HT(1B) receptor densities among these alcoholics, although the data must be considered as preliminary.

Aims: The Cloninger type 1 alcoholics are prone to anxiety, and in many cases patients have begun to use alcohol in order to relieve their anxiety. We have previously reported a decrease of the serotonin transporter density in the perigenual anterior cingulate cortex (pACC) in type 1 alcoholics. The 5-HT1A receptors are the binding sites for anxiolytic drug buspirone. We aimed to investigate the alteration in the density of 5-HT1A receptors, that may also alter the effect of serotonin in the pACC in alcoholics. Methods: The density of the serotonin receptor 5-HT1A among Cloninger type 1 and 2 alcoholics (nine and eight subjects, respectively) and 10 control subjects were determined by postmortem whole-hemisphere autoradiography with WAY-100635. Results: Substantially sparser 5-HT1A (by −31%, P = 0.010) density was observed in the pACC of alcoholic subjects in relation to non-alcoholic comparison subjects. In a secondary analysis for the difference between the alcoholic subtypes and controls, the 5-HT1A density was decreased significantly by −32% ( P = 0.015) in the upper level of pACC in type 1 alcoholics. Conclusions: The detected decrease of 5-HT1A receptor density on the pACC suggests further that the serotoninergic system is defected in the so-called affect region, especially in the type 1 alcoholics.

A number of studies have pointed to the importance of dopamine system in the context of alcoholism. Previous studies have shown lower dopamine transporter levels on late-onset Cloninger type 1 alcoholics. However, whether this lower level is due to a lower level of dopamine transporter protein or a lower level of dopaminergic nerve terminals remains unclear. The aim of this study was to compare putative alterations of dopaminergic terminals in caudate, putamen and nucleus accumbens of type 1 and type 2 alcoholics and healthy controls by using [ 3 H]dihydrotetrabenazine as a radioligand in postmortem human whole hemisphere autoradiography. We compared the present results with the findings of our earlier studies on the dopamine transporter in these same subjects, demonstrating that alcoholics do not differ significantly from controls in striatal [ 3 H]dihydrotetrabenazine binding. Although type 1 alcoholics have been reported to have up to 36% lower striatal dopamine transporter levels than controls, the results suggest that the density of their dopaminergic nerve terminals is not altered.

Indirect experimental evidence suggests that drugs acting on the alpha(2C)-adrenoceptor could be useful in the treatment of neuropsychiatric disorders such as depression and schizophrenia. In rodent brain, the highest levels of alpha(2C)-adrenoceptors are found in the striatum, with lower levels in cerebral cortex and hippocampus. In human brain, because of the poor subtype-selectivity of the available alpha(2)-adrenoceptor ligands, the localization of alpha(2C)-adrenoceptors has remained unknown. Recently, a selective alpha(2C)-adrenoceptor antagonist, JP-1302, was characterized, and to assess the presence of alpha(2C)-adrenoceptors in human brain, we performed competition binding in vitro receptor autoradiography with JP-1302 and the alpha(2)-adrenoceptor subtype nonselective antagonist [ethyl-(3)H]RS79948-197 on rat and human postmortem brain sections. In striatum of both species, JP-1302 vs. [ethyl-(3)H]RS79948-197 competition binding was biphasic, identifying high- and low-affinity binding sites, whereas in cortex and cerebellum, only low-affinity binding sites were detected. The results indicate that a significant portion of the alpha(2)-adrenoceptors in striatum is of the alpha(2C) subtype, whereas non-alpha(2C)-adreocneptors predominate in cortex and cerebellum. Because the alpha(2C)-adrenoceptor subtype distribution pattern appears to be conserved between rodents and humans, results obtained from studies on the role of the alpha(2C)-adrenoceptor in rodent models of neuropsychiatric disorders may be relevant also for human diseases.

Serotonin plays a role in the regulation of emotional states in amygdala which in turn affect the function of hypothalamus. The physiological effects of emotions are mediated to autonomic nervous system by the hypothalamus, also innervated by the serotonergic Raphe nuclei. Aims: We evaluated the putative alterations of the serotonin transporter (SERT) density in the paraventricular nucleus (PVN) of hypothalamus of Cloninger type 1 and 2 (early onset, anti-social) alcoholics and controls. Methods: The study was performed by human whole-hemisphere auto-radiography with ( 3 H)citalopram. Results: Substantially sparser SERT density (−26%) with a moderate effect size (0.53) was observed in the hypothalamus of alcoholic subjects in relation to non- alcoholic comparison subjects, although the result failed to reach statistical significance. In type 2 alcoholics, there was a trend towards decreased SERT binding with large effect size (0.88), and no correlation between the SERT binding and the age at the time of death. There was a strong positive correlation between the SERT binding in amygdala and in PVN in type 2 alcoholics (P = 0.001), and negative correlation in type 1 alcoholics (P = 0.05), and no correlation in the control subjects. The difference between the groups was significant (χ 2 = 16.75 ,P = 0.0002). Conclusions: Taken together, these preliminary results support the hypothesis that the serotonergic regulation in the hypothalamus and amygdala are defected especially in type 2 alcoholics.

Background: A dysfunction in brain serotonin turnover is a well-established factor associated with the impulsive and sociopathic behavior in alcoholics. The conjuncted alterations in functioning of serotonin transporter (SERT) may play a role in the regulation of emotional balance, judgement, and the adverse behavioral effects of ethanol misuse. These traits may be related to serotonergic regulation in the amygdala and prefrontal cortex. Methods: The binding of [3H]citalopram to SERT was evaluated in the amygdala of Cloninger type 1 and 2 alcoholics (n = 17), and nonalcoholic control subjects (n = 10) by postmortem whole-hemisphere autoradiography. Results: The SERT binding was substantially lower in the dorsal amygdala in alcoholic subjects when compared with the controls (−28%, effect size 1.26, P = 0.016). In secondary analysis, this reduction was observed in both alcoholic subgroups (−26% in type 1 alcoholics, and −33% in type 2 alcoholics). In ventral amygdala, no alteration was observed. There were significant correlations between the SERT binding in dorsal amygdala and in previous results from frontal cortical areas in alcoholics, depending on the type of alcoholic. Conclusions: These results suggest that SERT binding in the amygdala, as well as the differential regulation of the SERT in amygdala and frontal cortex in alcoholics may help to explain the dysfunctional regulation of emotions in alcoholics. Synapse 61:629–636, 2007. © 2007 Wiley-Liss, Inc.

Nucleus accumbens (NAC) is regulated by the dopaminergic and serotonergic pathways, and it is a brain area with a crucial role in the rewarding effects of ethanol. In this preliminary study, possible alterations of [3H]citalopram binding to serotonin transporter (SERT) were evaluated in the NAC of Cloninger type 1 and 2 alcoholics (nine and seven subjects, respectively), and nonalcoholic controls (10 subjects) by human postmortem whole-hemisphere autoradiography. The [3H]citalopram binding in the NAC was 35% higher in the alcoholics than in the controls; in the type 1 alcoholics, the binding was 54% and in the type 2 alcoholics it was 17% higher. Although the effect size showed medium effects (0.49-0.60), the results did not reach statistical significance due to large standard deviations. The [3H]citalopram binding declined significantly with age in the controls, but not in the alcoholics. In the controls, there was a significant positive correlation between the [3H]citalopram binding in the NAC and in the anterior cingulate gyrus, an area in which the [3H]citalopram binding has been shown to be lower among alcoholics. On the contrary, a significant negative correlation was observed in the type 2 alcoholics and no correlation in the type 1 alcoholics. In addition, there was a strong tendency toward a positive correlation between the SERT and dopamine transporter binding in the type 2 alcoholics, but not in the other groups. These preliminary results suggest a differential monoaminergic imbalance in type 1 and 2 alcoholism in brain areas important for the regulation of motivation, reward, and reinforcement.

A considerable number of human and animal studies have implied the importance of dopamine system and alterations in dopamine receptors in the context of alcoholism. However, it has remained unclear if the alcohol-abuse related dopaminergic deficit is specifically associated with certain receptor subtype. The aim of this study was to compare putative alterations of dopamine D(1) receptors in caudate and putamen of nine type 1 alcoholics, eight type 2 alcoholics and 10 healthy controls by using [(3)H]SCH 23390 as a radioligand in postmortem human whole hemisphere autoradiography. In addition, we compared the present results to our earlier studies on dopamine transporters and dopamine D(2) receptors in these same subjects and evaluated the putative correlations of dopamine D(1) receptor densities between the nucleus accumbens and the above-mentioned structures. Our results show that alcoholics do not have significantly different striatal dopamine D(1) receptor densities compared to controls. Neither were there any significant correlations between the dopamine D(1) receptors and the two other dopamine binding sites. However, the correlations of the dopamine D(1) receptors between nucleus accumbens and dorsal striatal structures were consistently and mostly statistically significantly positive in alcoholics, but not in controls, which may suggest some pathology related to addiction. In addition, considering the facts that dopamine D(1) receptors were more abundant in the mesolimbic nucleus accumbens than in the caudate or putamen and that there was a strong tendency towards lower binding among type 1 alcoholics may suggest the importance of dopamine D(1) receptors in reward and/or alcoholism.

The role of the dopamine (DA) system in brain reward mechanisms and the development of substance abuse has been well established. We review earlier animal and human studies on DA and alcoholism with some relevant issues relating to those studies. The present animal and human data suggest several alterations in the DA system in the context of alcoholism. Receptor studies imply that DA D(2) receptor density and function are lower at least among type 1 alcoholics, which suggests that they could benefit from drugs that enhance DAergic activity, such as partial DA agonists. These drugs could help to restore suboptimal levels of DAergic activity by reducing both the craving for alcohol in abstinence and the euphoria subsequent to alcohol's release of DA in the nucleus accumbens (NAC), thus providing negative reinforcement for relapse.

The distribution of the 5-HT7 receptor was analyzed using the selective 5-HT7 receptor antagonist radioligand [3H]SB-269970 and human brain whole hemisphere autoradiography. The results indicated that 5-HT7 receptors are most abundantly localized in the anterior thalamus and in the dentate gyrus. Other regions containing intermediate levels of 5-HT7 receptors included the hypothalamus, anterior cingulate gyrus, hippocampus, amygdala and certain brainstem nuclei. The distribution pattern obtained in this study supports the involvement of this receptor subtype in affective behavior and cognition.

Increasing evidence implies the involvement of the dopamine (DA) system in the pathogenesis of alcoholism. We measured striatal DA D(2) receptors in Cloninger type 1 and 2 alcoholics by using [(125)I]epidepride in human postmortem whole hemispheric autoradiography (WHA), which provides high-resolution images corresponding to positron emission tomographic (PET) studies. We also evaluated the correlation between transporter and receptor DA binding site densities and putative correlation of [(125)I]epidepride binding between the dorsal striatum and nucleus accumbens. In the type 1 alcoholics, the DA D(2) receptor density was 21.4-32.6% lower in all dorsal striatal structures (caudate, putamen, globus pallidus) when compared with the controls. Type 2 alcoholics had 19.6-21.4% lower binding in other dorsal striatal structures, except medial globus pallidus, where they were not significantly different from controls. The density of DA D(2) receptors and DAT had a significant positive correlation only in the putamen of type 1 alcoholics. The binding of [(125)I]epidepride showed also consistent and statistically significant positive correlation between nucleus accumbens and all dorsal striatal areas in type 1 alcoholics but not in the controls. In the type 2 alcoholics, the correlation was weaker than that observed in the type 1 alcoholics, and no correlation was observed between nucleus accumbens and globus pallidus. Our results show that these two subgroups of alcoholics have stark differences in their DA D(2) receptor binding characteristics. Type 2 alcoholics may have selective deficiency in the dorsal striatum, whereas in limbic structures they may not differ significantly from controls. Moreover, WHA provides a useful tool for detailed mapping of neuronal receptors in healthy as well as diseased brain, and can also be used in radioligand development for PET.

It is generally agreed that there is a deterioration in brain dopamine (DA) system with aging. The role of the mesolimbic DA in brain ethanol reinforcement is well established, with nucleus accumbens (NAC) serving as a major terminal area of this system, whereas dorsal striatum is more associated with motor control. The aim of this study was to compare putative age-related alterations of dopamine transporters (DAT) in dorsal and ventral striatum of healthy controls and alcoholics. We studied the effect of age on DAT in caudate (NC), putamen (Pu), and nucleus accumbens (NAC) of eight type 1 and 2 alcoholics and 10 healthy controls by using [(125)I]PE2I as a radioligand for postmortem human whole hemisphere autoradiography. In the type 1 alcoholic group age and DAT density did not correlate significantly with any of the structures studied. The mean densities of DAT declined significantly with age in controls and type 2 alcoholics in dorsal striatum (NC, Pu) (range of correlation coefficient from -0.49 to -0.94), but not statistically significantly in NAC. In type 1 alcoholics the lack of correlation between DAT density and age may indicate a preexisting dopaminergic deficit in this patient group, whereas age-related decline among type 2 alcoholics resembled that of healthy controls. Furthermore, dorsal striatal DAT may be more vulnerable to age-related decline than DAT in NAC. This is supported by the notion that DAT in NAC and dorsal striatum have different molecular weights.

Lesions of the medial prefrontal cortex and dysfunctions in serotonin turnover are two well-established factors associated with impulsive and sociopathic behaviors, but no firm neuroanatomical data have linked these pathophysiological findings. The aims of this study were to identify putative areas in the human brain that are rich in serotonin transporter sites, particularly within the medial prefrontal cortex, and to determine whether serotonin transporter density in this area is altered among alcoholic subjects.Serotonin transporter density was measured among 17 alcoholic and 10 nonalcoholic comparison subjects by postmortem whole-hemisphere autoradiography with [(3)H]citalopram.In the human cerebral cortex, serotonin transporter binding sites were concentrated in the perigenual anterior cingulate cortex. Substantially sparser serotonin transporter density (up to 35%) was observed in the perigenual anterior cingulate cortex of alcoholic subjects in relation to nonalcoholic comparison subjects. After adjustment for age and postmortem delay, this finding remained statistically significant.A lower serotonin transporter density among the alcoholic subjects was observed, specifically in the so-called "affect" region, suggesting an association between ethanol addiction and dysfunctional serotonergic neurotransmission in this area.

Dopaminergic mechanisms are involved in the positive reinforcing and addicting effects of alcohol. Positron emission tomography (PET) and single photon emission tomography (SPET) studies have indicated alterations in striatal dopamine transporters (DAT) and in presynaptic dopamine (DA) function in alcoholics, although also contradictory results have been reported. Normal variations in blood flow, metabolism, and receptor densities are apparently important to brain function. Such variations are known to decrease during pathophysiological processes, such as epilepsy, whereas normal receptor distributions are broadly heterogenous. We evaluated the densities and heterogeneities of striatal DAT in 8 adult-onset, Cloninger type I alcoholics and 10 controls using [125I]N-(3-iodoprop-2E-enyl)-2beta-carbomethoxy-3beta- (4'-methylphenyl)nortropane ([125I]PE2I) as a ligand for human postmortem whole hemisphere autoradiography, which provided high resolution images of the brain when compared with in vivo PET and SPET. The mean density and heterogeneity of DAT were markedly lower in the alcoholics. A significant linear correlation existed between DAT density and heterogeneity, as well as between DAT densities in the nucleus accumbens and in the dorsal striatum (caudate and putamen) in alcoholics, but not consistently in controls. The observed low DAT density and heterogeneity in the dorsal striatum suggest that type 1 alcoholics may have a dysfunctional DA system. These data indicate that human whole hemisphere autoradiography with the analysis of binding heterogeneity may be a relevant tool to measure pathological processes in the brain.

Much research interest has lately been focused on the dopamine transporter function in brain. Recent findings indicate that dopamine reuptake is more like a highly regulated than a constitutive determinant of dopamine clearance. Positron emission tomography (PET) and single-photon emission tomography (SPET) offer unique methods to study dopamine transporter function. Results from in vivo PET and SPET studies correspond well with in vitro studies performed on post mortem human brain tissue. Considering some of the variances between in vitro and in vivo receptor binding phenomena it may be that the role of a compound to alter binding to monoamine uptake sites in vitro does not indicate its potential to affect monoamine transporters after administration in vivo. This discrepancy may be better understood taking into account recent studies indicating the possibility of a rapid regulation of transporter function and surface expression. Furthermore, the dopamine transporter is a fruitful target for CNS drug discovery. Fundamental nature of drug actions in vivo may be studied using demonstrated in vitro and in vivo imaging methods.

Alcohol acts through mechanisms involving the brain neurotransmitter dopamine (DA) with the nucleus accumbens as the key zone for mediating these effects. We evaluated the densities of DA D(2)/D(3) receptors and transporters in the nucleus accumbens and amygdala of post-mortem human brains by using [(125)l]epidepride and [(125)I]PE2I as radioligands in whole hemispheric autoradiography of Cloninger type 1 and 2 alcoholics and healthy controls. When compared with controls, the mean binding of [(125)I]epidepride to DA D(2)/D(3) receptors was 20% lower in the nucleus accumbens and 41% lower in the amygdala, and [(125)I]PE2I binding to DA transporters in the nucleus accumbens was 39% lower in type 1 alcoholics. These data indicate that dopaminergic functions in these limbic areas may be impaired among type 1 alcoholics, due to the substantially lower number of receptor sites. Our results suggest that such a reduction may result in the chronic overuse of alcohol as an attempt to stimulate DA function.

γ-Aminobutyric acid type A (GABA A ) receptor ionophore ligand t -[ 35 S]butylbicyclophosphorothionate ([ 35 S]TBPS) was used in an autoradiographic assay on brain cryostat sections to visualize and characterize atypical GABA-insensitive [ 35 S]TBPS binding previously described in certain recombinant GABA A receptors and the cerebellar granule cell layer. Picrotoxinin-sensitive but 1-mM GABA-insensitive [ 35 S]TBPS binding was present in the rat cerebellar granule cell layer, many thalamic nuclei, subiculum and the internal rim of the cerebral cortex, amounting in these regions up to 6% of the basal binding determined in the absence of exogenous GABA. Similar binding properties were detected also in human and chicken brain sections. Like the GABA-sensitive [ 35 S]TBPS binding, GABA-insensitive binding was profoundly decreased by pentobarbital, pregnanolone, loreclezole and Mg 2+ . The binding was reversible and apparently dependent on Cl − ions. Localization of the GABA-insensitive [ 35 S]TBPS binding was not identical to that of high-affinity [ 3 H]muscimol binding and diazepam-insensitive [ 3 H]Ro 15-4513 binding, two previously established receptor subtype-dependent binding heterogeneities in the rat brain. The present study reveals a component of the GABA-ionophore enriched in the thalamus and cerebellar granule cells, possibly representing poorly desensitized or desensitizing receptors.

The spatial pattern of striatal dopamine transporter density in the living human brain was tested by duplicate SPECT scans with , or and striatal phantom measurements. The resolution-dependent spatial variation was calculated by the fractal analysis of SPECT images. This variation, which depends on the size of the region of interest, was described by the spatial dispersion i.e. the standard deviation of the count densities divided by the mean density. In each sub-region, the observed and methodological dispersions were computed, and the resulting spatial dispersion was calculated. The methodological dispersion is caused by the imaging resolution, flood field non-uniformity, count density, scatter, reconstruction errors and partial volume effects, whereas the spatial dispersion is based on the cerebral heterogeneity of the dopamine transporter density. Recognition of the normal variation in heterogeneity is important in evaluation of the striatal dopamine transporter density between controls and patients suffering from various neuropsychiatric disorders. Keywords: fractal, heterogeneity, receptor, SPECT, striatum

Accumulating evidence continues to link certain aspects of the endogenous cannabinoid (EC) system with alcohol dependence, negative-reinforcement learning, and the modulation of stress responses. Specific alterations in brain regions that are related to stress and negative-reinforcement learning have been reported to exist in Cloninger type 1 and type 2 alcoholics. To study possible differences in profiles of EC systems between Cloninger type 1 (n = 9) and type 2 (n = 8) alcoholics and non-alcoholic control subjects (n = 10), we analyzed post-mortem amygdala and hippocampus brain samples for several ECs by quantitative liquid chromatography with triple quadrupole mass-spectrometric detection. A significant difference was found between these 3 groups in terms of EC profiles in the amygdala (p = 0.037). In particular, this difference was prominent for variations in docosahexaenoylethanolamide levels, which were significantly higher in type 1 alcoholics (p = 0.022) when compared to controls. There was also a large negative correlation between anandamide concentration and mGlu1/5 receptor density in the hippocampi of Cloninger type 1 alcoholics (R = −0.88, p = 0.002), which was not seen in Cloninger type 2 alcoholics or in controls. Although preliminary, and from relatively small diagnostic groups, these results suggest that the EC system profile may be altered in the hippocampus and amygdala of Cloninger type 1 alcoholics., final draft, peerReviewed

The 5-HT(1B) receptor has been associated with alcohol dependence, impulsive or alcohol-related aggressive behavior, and anxiety. The aim of this study was to determine whether or not the 5-HT(1B) receptor density differs in brain samples from anxiety-prone Cloninger type 1 alcoholics and socially hostile, predominantly male, type 2 alcoholics, and controls. Whole-hemispheric 5-HT(1B) receptor density was measured in eight regions of postmortem brains from 17 alcoholics and 10 nonalcoholic controls by autoradiography with tritiated GR-125743 and unlabeled ketanserin to prevent 5-HT(1D) binding. The 5-HT(1B) receptor density was not altered significantly in any of the studied regions. However, some correlations were observed in types 1 and 2 alcoholics only. The 5-HT(1B) receptor density decreased with age in type 1 alcoholics only. There was a significant positive correlation between 5-HT(1B) receptor and serotonin transporter densities in the head of caudate of type 1 alcoholics only. There was a significant positive correlation between 5-HT(1B) receptor density and dopaminergic terminal density, as estimated by vesicular monoamine transporter 2 measurement in the nucleus accumbens of type 2 alcoholics only. There were no significant correlations between 5-HT(1B) receptor and dopamine transporter or dopamine D2/D3 receptor densities in any of the subject groups. In conclusion, these results do not indicate primary changes in 5-HT(1B) receptor densities among these alcoholics, although the data must be considered as preliminary.

The endogenous cannabinoid (EC) system has been recently implicated in several neuropsychiatric disorders. This study analyzed post-mortem brain regions of Cloninger type 1 (n = 9) and 2 (n = 8) alcoholics and non-alcoholic controls (n = 10) for ECs by quantitative liquid chromatography with triple quadrupole mass spectrometric detection. A significant difference was found in anandamide (AEA) levels in nucleus accumbens (NAcc) between the three groups (p = 0.047). AEA levels were significantly lower when compared to controls in both perigenual anterior cingulate (p = 0.017) and frontal cortices (p = 0.018) of type 1 alcoholics. Similar trends were observed for dihomo-γ-linolenoyl ethanolamide and docosahexaenoyl ethanolamide, but not for 2-arachidonoylglycerol, palmitoyl ethanolamide, or oleoyl ethanolamide. Although preliminary, and from diagnostic groups with a relatively small number of subjects and substantially different mean ages for each group, these results suggest that the EC system may be hyperactive in type 2 alcoholics and hypoactive in type 1 alcoholics.

The iodine-123 labelled selective ligand N-(3-iodoprop-2E-enyl)-2beta-carbomethoxy-3beta-(4-methylphe nyl)nortr opane ([(123)I]PE2I) has been developed and has been shown to be suitable for single-photon emission tomography imaging of the dopamine transporter. In this study the influence of age on ligand binding was investigated in 16 healthy males with an age range of 23- 75 years. Single-photon emission tomography (SPET) imaging was performed with a triple-headed gamma camera. A simplified reference region model, in which the input function was derived from the non-displaceable cerebellar compartment, was used to calculate the volume of distribution in the striatum. The volume of distribution was shown to decline with age (-0.4%/year; P0.005). The results were in agreement with in vivo and in vitro findings of a decline in dopamine transporter binding with age. The findings confirm the suitability of [(123)I]PE2I for SPET imaging in clinical routine but emphasize the necessity of using age-matched controls in patient studies.

Prolyl oligopeptidase (POP) is a serine endoprotease that hydrolyses peptides shorter than 30-mer. POP may have a role in inositol 1,4,5-triphosphate (IP(3)) signaling and in the actions of antidepressants, and POP inhibitors have exhibited antiamnesic and neuroprotective properties. However, little is known about the distribution of POP protein in the brain. We used immunohistochemistry to localize POP enzyme in the human whole hemisphere and in the rat whole brain. In humans, the highest POP densities were observed in caudate nucleus and putamen, hippocampus and cortex. In the rat, the highest POP densities were found in substantia nigra, hippocampus, cerebellum and caudate putamen. In general, the distribution of POP in human and rat brains was very similar and resembled that of IP(3) receptors. Our findings are support for a role of POP in movement regulation, cognition and possibly in IP(3) signaling. The expression of POP in processing nuclei further supports its function beyond neuropeptide metabolism.

Dorsal striatum is regulated by the serotonergic system, and it is a brain area with a role in the development of obsessive thought patterns, which may be related to addiction. In this study, possible alterations of [(3)H]citalopram binding to serotonin transporter (SERT) were evaluated in the dorsal striatum of Cloninger type 1 and 2 alcoholics, and nonalcoholic control subjects by postmortem whole-hemisphere autoradiography in humans. The SERT binding was significantly lower (-26%, effect size 1.74) in the caudate body of alcoholics. The SERT binding tended to be lower also in the other parts of the dorsal striatum in alcoholics, but the results did not reach significance. In addition, there was a significant positive correlation, especially in type 1 alcoholics, between the SERT binding in the body of the caudate and in the perigenual anterior cingulate cortex, an area in which the SERT binding has been shown to be lower among alcoholics. These results give preliminary evidence to suggest that the SERT binding in the dorsal striatum may be lower in alcoholics, and that the serotonergic system may be affected in cortical and striatal areas simultaneously. The cortico-striatal-thalamic axis may have an important role in fully developed addictions and the characterization of these correlations within the serotonergic system may lead to a better understanding of the anatomical dynamics underlying the neurochemistry of alcoholism.

Objective We tested the hypothesis that topiramate is more effective than placebo in reducing symptoms in patients with treatment-resistant schizophrenia when combined with ongoing antipsychotic medication. Method Twenty-six hospitalized treatment-resistant patients with chronic DSM-IV-diagnosed schizophrenia participated in a randomized, double-blind, placebo-controlled trial in which 300 mg/day of topiramate was gradually added to their ongoing treatment (clozapine, olanzapine, risperidone, or quetiapine) over two 12-week crossover treatment periods. Data were collected from April 2003 to November 2003. Results In intention-to-treat analysis, topiramate was more effective than placebo in reducing Positive and Negative Syndrome Scale general psychopathologic symptoms (effect size = 0.7, p = .021), whereas no significant improvement was observed in positive or negative symptoms. Conclusion Glutamate antagonist topiramate may be an effective adjuvant treatment in reducing general psychopathologic symptoms in patients with schizophrenia resistant to treatment with second-generation antipsychotics.

We measured cortical dopamine transporter (DAT) in Cloninger type 1 and 2 alcoholics by using [ 125 I]PE2I as a radioligand in human postmortem whole hemispheric autoradiography, and evaluated the putative correlations of DAT between cortical areas and nucleus accumbens. There was a low, but distinct cortical binding in the cryosections. The mean binding was generally higher in both groups of alcoholics compared to controls, and the results reached statistical significance with a large effect size (1.25) in the temporal cortex of type 2 alcoholics. This is surprising, because several studies have reported lower DAT densities in the striatum among alcoholics compared to controls. Moreover, the density of DAT had a statistically significant positive correlation between temporal cortex and nucleus accumbens in controls, whereas among type 2 alcoholics the correlation was statistically significantly negative, which may suggest some pathology relating to the antisocial behaviour of these alcoholics.

A large body of evidence indicates the importance of dopamine (DA) activation for ethanol reinforcement, and animal models of alcoholism have implied the involvement of DA D 1 receptors in this context. We studied cortical DA D 1 receptors in nine type 1 alcoholics (late-onset, binge-drinker), eight type 2 alcoholics (early-onset, antisocial) and 10 controls by using [ 3 H]SCH23390 as a radioligand in postmortem human whole hemisphere autoradiography. We also evaluated correlations of DA D 1 receptors between the cortical and subcortical areas and between cortical DA transporters and DA D 2 and D 3 receptors by comparing the present results to our earlier studies. On the average, type 2 alcoholics were younger and had more violent causes of death than type 1 alcoholics and controls. There were no statistically significant differences between the groups, suggesting that cortical DA D 1 receptors do not play a major role in alcoholism. However, among type 2 alcoholics, the binding was consistently lower (8.6%–22.3%) than among controls, and the effect sizes showed a large effect in the anterior cingulate (0.90) and frontal (0.87) cortices. Interestingly, among type 2 alcoholics, the correlation of DA D 1 receptors between two ventral midbrain structures (substantia nigra and amygdala) and anterior cingulate cortex was significantly negative, whereas in the type 1 alcoholics and controls, the correlations were significantly positive.

Alcoholism has been associated with lower density of striatal dopamine (DA) D2 receptors, but there is much less data on cortical DA D2 receptors. We evaluated the [125I]epidepride binding to DA D2 receptors in Cloninger type 1 and 2 alcoholics and controls in frontal, temporal, and anterior cingulate cortices by using human postmortem whole hemispheric autoradiography, which provides high-resolution images corresponding to positron emission tomographic (PET) studies. Type 1 alcoholics had lower and type 2 alcoholics had higher DA D2 receptor density in all cortical areas compared to controls. Although the results did not reach statistical significance, the effect sizes were high. The DA D2 receptor density in type 2 alcoholics decreased statistically significantly with age, and after correcting for age the binding values also fell below the level of controls. A statistically non-significant tendency towards a decrease of cortical DA D2 receptors was seen in controls, whereas in the type 1 alcoholic group no consistent correlation or even tendency towards increase with age was observed. Our results give preliminary evidence that DA D2 receptors in cortical areas may be lower among both groups of alcoholics, but not necessarily of same magnitude as in subcortical structures. The rapid decline of cortical DA D2 receptors among type 2 alcoholics may have some relevance to their antisociality, because this trait tends to diminish with age. The absence of correlation or even tendency towards increase of cortical DA D2 receptors with age seen in type 1 alcoholics may give further evidence that they have a pre-existing dopaminergic deficit. However, these results especially regarding aging effect must be considered as preliminary due to the different age-range of type 2 alcoholics compared to two other groups. Synapse 54:129–137, 2004. © 2004 Wiley-Liss, Inc.

The distribution of the 5-HT7 receptor was analyzed using the selective 5-HT7 receptor antagonist radioligand [3H]SB-269970 and human brain whole hemisphere autoradiography. The results indicated that 5-HT7 receptors are most abundantly localized in the anterior thalamus and in the dentate gyrus. Other regions containing intermediate levels of 5-HT7 receptors included the hypothalamus, anterior cingulate gyrus, hippocampus, amygdala and certain brainstem nuclei. The distribution pattern obtained in this study supports the involvement of this receptor subtype in affective behavior and cognition.

Increasing evidence implies the involvement of the dopamine (DA) system in the pathogenesis of alcoholism. We measured striatal DA D(2) receptors in Cloninger type 1 and 2 alcoholics by using [(125)I]epidepride in human postmortem whole hemispheric autoradiography (WHA), which provides high‐resolution images corresponding to positron emission tomographic (PET) studies. We also evaluated the correlation between transporter and receptor DA binding site densities and putative correlation of [(125)I]epidepride binding between the dorsal striatum and nucleus accumbens. In the type 1 alcoholics, the DA D(2) receptor density was 21.4–32.6% lower in all dorsal striatal structures (caudate, putamen, globus pallidus) when compared with the controls. Type 2 alcoholics had 19.6–21.4% lower binding in other dorsal striatal structures, except medial globus pallidus, where they were not significantly different from controls. The density of DA D(2) receptors and DAT had a significant positive correlation only in the putamen of type 1 alcoholics. The binding of [(125)I]epidepride showed also consistent and statistically significant positive correlation between nucleus accumbens and all dorsal striatal areas in type 1 alcoholics but not in the controls. In the type 2 alcoholics, the correlation was weaker than that observed in the type 1 alcoholics, and no correlation was observed between nucleus accumbens and globus pallidus. Our results show that these two subgroups of alcoholics have stark differences in their DA D(2) receptor binding characteristics. Type 2 alcoholics may have selective deficiency in the dorsal striatum, whereas in limbic structures they may not differ significantly from controls. Moreover, WHA provides a useful tool for detailed mapping of neuronal receptors in healthy as well as diseased brain, and can also be used in radioligand development for PET. Hum. Brain Mapp. 20:91–102, 2003. © 2003 Wiley‐Liss, Inc.

The role of the dopamine system in brain reward mechanisms and development of substance abuse is well-established with nucleus accumbens as a key structure in mediating these effects. Several studies on alcoholism have indicated defects in dopaminergic neurotransmission and alterations in dopamine receptor densities. However, it has remained unclear if the substance abuse-related dopaminergic defect is specifically associated with a certain receptor subtype. The aim of this study was to compare putative alterations of dopamine D(1,) D(2), and D(3) receptors in nucleus accumbens, amygdala, and substantia nigra among alcoholics and controls. We studied the densities of dopamine D(1) and D(3) receptors in brains of 9 type 1 alcoholics, 8 type 2 alcoholics, and 10 healthy controls by using postmortem human whole hemisphere autoradiography. The mean densities of dopamine D(1) and D(3) receptors were at the same level in all study groups. Combining these with our previous results, our data suggest that among type 1 alcoholics dopamine transporters are lower in nucleus accumbens and dopamine D(2), but not D(1) or D(3) receptors in nucleus accumbens and amygdala. Further, the densities of all these dopamine-binding sites among type 2 alcoholics are at the level of healthy controls. The results suggest that lower dopamine receptor density is specific for D(2) receptor and for type 1 alcoholism, which supports Cloninger's neurogenetic model of two alcoholic subtypes, and indicates the importance of classifying these subgroups separately when issues related to dopaminergic activity are studied.

Brain neurotransmitter and receptor systems are altered in animals exhibiting increased ethanol preference; most studies being on brain dopamine and 5-hydroxytryptamine(5HT) systems. The role of histaminergic systems in high ethanol consumption is not well understood. It has been suggested that histaminergic tuberomamillary neurons participate in the inhibitory control of reward functions [1]. Lesions of dopaminergic neurons in the substantia nigra have been shown to disrupt reward-related processes [2]. Thus, it is possible that dopaminergic nigrostriatal and histaminergic tuberomamillary striatal systems may act in a reciprocal fashion to regulate reward functions. Pathophysiological differences in the rat brain histaminergic system have been studied on selectively-bred alcohol-preferring AA rats versus their alcohol-avoiding ANA counterparts. Histamine and its metabolite tele-methylhistamine were measured in brains of these rats. Both have been found in higher concentrations in alcohol-preferring rats [3]. Here we studied if similar changes could be detected in humans abusing alcohol.

Late onset type 1 alcoholism has been suggested to be associated with decreased dopaminergic transmission. Our hypothesis was that late onset type 1 alcoholics have also abnormal extrastriatal dopamine D(2)/D(3) receptor distribution. We performed binding, heterogeneity and laterality analysis of extrastriatal and striatal dopamine D(2)/D(3) receptors in nine late onset male alcoholics and in 12 age-matched healthy males. A radioligand, [(123)I]epidepride was used in high resolution single-photon emission tomography (SPET). Specific binding of epidepride in the left temporal pole was significantly (P

Neuroleptics are known to cause anhedonia and attenuate sexual behaviour at therapeutic doses in humans. These effects are assumed to result from the dopamine antagonism of the drugs. It has been observed that a mixed dopamine D1/D2 antagonist, haloperidol, may cause a reduction in the number of intromissions required to achieve ejaculation. On the other hand, dopamine antagonists are considered unable to modify sexual behaviour once the copulatory sequence is initiated. In this study, male rats received low doses of haloperidol (30 or 60 microg/kg) before the investigation of sexual behaviour in five consecutive days and the mating test was repeated after withdrawal periods of four and five days. Haloperidol dose-dependently reduced intromission frequency, and this effect was maintained for four days after withdrawal. Ejaculation latency was reduced in all groups, including controls. The results indicate that at low doses haloperidol dose-dependently reduces intromission frequency, and the effect of a repeated dosage may persist several days after cessation of medication.

The iodine-123 labelled selective ligand N-(3-iodoprop-2E-enyl)-2-beta-carbomethoxy-3beta-(4-methylphenyl) nortropane ([123I]PE2I) was evaluated as a probe for in vivo dopamine transporter imaging in the human brain. Six healthy subjects were imaged with a high-resolution single-photon emission tomography scanner. Striatal radioactivity peaked at 1 h after injection. The background radioactivity was low. The volume of distribution in the striatum was 94+/-24 ml/ml. The results were compared with those of [123I]beta-CIT imaging. There was no significant uptake of [123I]PE2I in serotonin-rich regions such as the midbrain, hypothalamus and anterior gingulus, suggesting that in vivo binding is specific for the dopamine transporter. One main polar metabolite of [123I]PE2I was found in plasma, and the parent plasma concentration decayed rapidly. Radiation exposure to the study subject is 0.022+/-0.004 mSv/MBq (effective dose). The preliminary results suggest that [123I]PE2I is a selective SPET ligand for imaging striatal dopamine transporter density.

The dopaminergic system is critically involved in reward mechanisms mediating the reinforcing effects of ethanol (di Chiara and Imperato 1988). However, it has remained unclear if the alcohol-abuse-related dopaminergic deficit is specifically associated with a certain dopamine (DA) receptor subtype. One of these DA D1–D5 receptors (Hartman and Lanau 1997), the DA D3 receptor, is expressed primarily in the regions of the brain that are thought to influence cognitive, emotional and behavioural function (Sokoloff and Schwartz 1995). However, the role of the DA D3 receptor in alcoholics has been difficult to study because of its low abundance and the lack of selective ligands, and to our knowledge, there is only one published study on the issue (Tupala et al. 2003a). The aim of this study was to compare DA D3 receptors in the anterior cingulate and frontal and temporal cortices of nine type 1 alcoholics, eight type 2 alcoholics and ten healthy controls by using [H]PD 128907 as a radioligand in postmortem human whole-hemisphere autoradiography. The procedure has been previously described in detail (Tupala et al. 2003a). Type 1 alcoholics are characterised by late onset, social dependency and alternating binges and abstinence, whereas type 2 alcoholics are characterised by early onset with impulsive and antisocial behaviour (Cloninger 1995). [H]PD 128 907 [(+)-(4aR,10bR)-3,4,4a, 10b-tetrahydro-4-propyl-2H,5H-(1)benzopyrano-(4,3b)-1, 4-oxazin-9-ol] binds specifically with a high affinity to DA D3 receptors, with 18to 40-fold selectivity for DA D3 over D2 receptors (Pugsley et al. 1995), whereas the more commonly used older ligand [H]7-OH-DPAT has only 7to 24fold selectivity using different assay conditions (Gonzalez and Sibley 1995; Pugsley et al. 1995). In addition, because we have previously shown striatal DA transporter (DAT) binding to correlate between the mesolimbic nucleus accumbens (NAC) and dorsal striatum (caudate and putamen) and a similar phenomenon has been shown in one singlephoton emission tomography (SPET) study in type 1 alcoholics, but not in controls (Repo et al. 1999; Tupala et al. 2001), we also evaluated the correlations of [H]PD 128707 binding between the cortical and subcortical areas by comparing the present results with our earlier studies on the NAC and substantia nigra (SN) to test whether the relative individual levels of these receptors are maintained in different brain areas in these three groups. In addition, we compared the present results with our earlier findings on cortical DA D2 receptors with [ I]epidepride in these same subjects (Tupala et al. 2004). Our results show that, in spite of the very low densities, it is possible to visualise DA D3 receptors in cortical areas of human whole-hemisphere cryosections. The DA D3 receptors were at the same level in all groups (1.75–2.33± 0.42–0.81 fmol/mg, mean±SD) and no statistically significant differences were found. In addition, the calculated effect sizes (Cohen 1988) were also quite low, thus supporting the results. There was some tendency towards posE. Tupala (*) Department of Forensic Psychiatry, Niuvanniemi Hospital, University of Kuopio, 70240 Kuopio, Finland e-mail: erkki.tupala@niuva.fi Tel.: +358-44-5597317 Fax: +358-17-203494

We have shown that alcoholic patients have a lower number of dopamine transporters in the nucleus accumbens, which mediates the rewarding effects of addictive drugs. Thus, certain dopaminergic agents may be beneficial in the treatment of alcohol withdrawal and in the long-term treatment of alcoholism with selective use.