AMPA receptors in post-mortem brains of Cloninger type 1 and 2 alcoholics: a whole-hemisphere autoradiography study.

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, [(3)H] 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 [(3)H] 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 [(3)H] 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.

Endogenous cannabinoids in amygdala and hippocampus in post-mortem brains of Cloninger type 1 and 2 alcoholics.

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.

Decreased GABA(A) benzodiazepine binding site densities in postmortem brains of Cloninger type 1 and 2 alcoholics.

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.

mGluR1/5 receptor densities in the brains of alcoholic subjects: a whole-hemisphere autoradiography study.

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.

Whole-hemisphere autoradiography of 5-HT1B receptor densities in postmortem alcoholic brains.

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.

Endogenous cannabinoids in post-mortem brains of Cloninger type 1 and 2 alcoholics.

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-gamma-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.

5-HT(1A) receptors in the frontal cortical brain areas in Cloninger type 1 and 2 alcoholics measured by whole-hemisphere autoradiography.

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-HT(1A) receptors are the binding sites for anxiolytic drug buspirone. We aimed to investigate the alteration in the density of 5-HT(1A) receptors, that may also alter the effect of serotonin in the pACC in alcoholics. METHODS: The density of the serotonin receptor 5-HT(1A) 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-HT(1A) (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-HT(1A) 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-HT(1A) 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.

Striatal dopaminergic terminals in type 1 and type 2 alcoholics measured with [3H]dihydrotetrabenazine and human whole hemisphere autoradiography.

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.

Autoradiographic characterization of alpha(2C)-adrenoceptors in the human striatum.

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.

Cortical dopamine D(1) receptors in type 1 and type 2 alcoholics measured with human whole hemisphere autoradiography.

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.

Correlation between the SERT binding densities in hypothalamus and amygdala in Cloninger type 1 and 2 alcoholics.

UNLABELLED: 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 [3H]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 (chi2 = 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.

Amygdala serotonin transporters in alcoholics measured by whole hemisphere autoradiography.

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 [(3)H]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.

Distribution of immunoreactive prolyl oligopeptidase in human and rat brain.

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.

Visualization of the cortical dopamine transporter in type 1 and 2 alcoholics with human whole hemisphere autoradiography.

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.

Lower serotonin transporter binding in caudate in alcoholics.

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.

Nucleus accumbens serotonin transporters in alcoholics measured by whole-hemisphere autoradiography.

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.

Topiramate add-on in treatment-resistant schizophrenia: a randomized, double-blind, placebo-controlled, crossover trial.

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.

Striatal dopamine D1 receptors in type 1 and 2 alcoholics measured with human whole hemisphere autoradiography.

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.

Dopamine and alcoholism: neurobiological basis of ethanol abuse.

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.