Trazodone effects on [H]-paroxetine and alpha(2)-adrenoreceptors in platelets of patients with major depression.

Trazodone is an antidepressant which behaves as a selective 5-HT(2) antagonist and 5-HT reuptake inhibitor. The lack of information on its effects in vivo prompted us to evaluate alpha(2)-adrenoceptors by means of the specific binding of [(3)H]-rauwolscine, and the 5-HT transporter (SERT) by means of the binding of [(3)H]-paroxetine ([(3)H]-Par), in platelets of depressed patients, before and after one month of treatment with trazodone (75-300 mg/day). Twenty-five outpatients of both sexes with a diagnosis of major depression, as assessed by the Structured Clinical Interview for DSM IV, were included in the study. Depressive symptoms were evaluated by means of the Hamilton Rating Scale for Depression: the total score (mean +/- SD) was 20 +/- 6 at baseline (t(0)) and 7 +/- 4 after one month of treatment (t(1)). Platelet membranes, [(3)H]- rauwolscine and [(3)H]-Par bindings were carried out according to standardized protocols. The results showed that the B(max) values of [(3)H]-Par were statistically lower at t(1) than at t(0) (733 +/- 30 vs 1471 +/- 99, P < 0.001), while the K(d) and the [(3)H]-rauwolscine binding parameters remained unchanged. The findings of this study suggest that in vivo trazodone modifies the number of the SERT proteins and that, perhaps, most of its antidepressant properties are related to this activity.

Male axillary extracts modify the affinity of the platelet serotonin transporter and impulsiveness in women.

The presence of functional pheromones in axillary extracts in humans is still matter of debate. Scattered data suggest that unidentified human axillary compounds with pheromonal activity may influence mood and this may occur, perhaps, through the modulation of the serotonin (5-HT) system that has been linked to mood by several findings. Therefore, the aim of this study was to assess the possible changes of a peripheral marker of the 5-HT system, i.e., the platelet 5HT transporter, and of some psychological tests, in a group of women who were exposed to male axillary extracts (group 1). A matched group of women who underwent an exposure to a neutral solution, were used as control subjects (group 2). The 5-HT transporter was evaluated by means of the specific binding of (3)H-paroxetine ((3)H-Par) to platelet membranes, as well as by means of (3)H-5-HT reuptake in whole platelets, at baseline (T0) and 1h after the stimulation (T1). The following tests were used: the "Experiences in Close Relationships" questionnaire (ECR), the latest version of the Barratt Impulsiveness Scale (BIS-11) and the Structured Clinical Interview for Mood Spectrum, self-reported version. The dissociation constant (Kd) of (3)H-Par binding showed a significant decrease at T1 only in the women exposed to male axillary extracts, as compared with baseline values, while the Bmax and (3)H-5-HT reuptake parameters did not show any change in both groups. The correlation analyses showed that at T0, the Kd values correlated significantly and positively with the factor of motor impulsiveness in all subjects. Two factors of the BIS-11, in particular, the attentional and the motor impulsiveness were significantly lower at T1 in the group 1. Further, at T1 and still in the group 1, a significant and positive correlation was measured between the Kd values and two ECR attachment styles, the secure and preoccupied, as well as with the ECR anxiety scale. Taken together, these findings suggest that the application of male axillary extracts to women may modify the affinity of their platelet 5-HT transporter, as well as of some impulsiveness and romantic attachment characteristics. The substances responsible for this effect remain to be identified.

Impulsivity, gender, and the platelet serotonin transporter in healthy subjects.

The present study explored the possible relationships between impulsivity, gender, and a peripheral serotonergic marker, the platelet serotonin (5-HT) transporter (SERT), in a group of 32 healthy subjects. The impulsivity was measured by means of the Barratt Impulsivity Scale, version 11 (BIS-11), a widely used self-report questionnaire, and the platelet SERT was evaluated by means of the specific binding of (3)H-paroxetine ((3)H-Par) to platelet membranes, according to standardized protocols. The results showed that women had a higher BIS-11 total score than men, and also higher scores of two factors of the same scale: the motor impulsivity and the cognitive complexity. The analysis of the correlations revealed that the density of the SERT proteins, as measured by the maximum binding capacity (B(max)) of (3)H-Par, was significantly and positively related to the cognitive complexity factor, but only in men. Men showed also a significant and negative correlation with the dissociation constant, Kd, of ((3)H-Par) binding, and the motor impulsivity factor. These findings suggest that women are generally more impulsive than men, but that the 5-HT system is more involved in the impulsivity of men than in that of women.

Latest advancements on serotonin and dopamine transporters in lymphocytes.

Different data show that circulating lymphocytes possess functional serotonin and dopamine transporters (SERT and DAT, respectively). This papers aims to review most of the available literature on this topic, while highlighting the possible role of SERT and DAT, as well as that of their substrates including antidepressants on the immune system.

Alterations of the dopamine transporter in resting lymphocytes of patients with different psychotic disorders.

The aim of our study was to investigate and compare the dopamine (DA) transporter (DAT) in resting lymphocytes of 20 psychotic patients and 20 healthy control subjects, by means of both the binding parameters (Bmax and Kd) of 3H-WIN 35,428, and the reuptake parameters (Vmax and Km) of 3H-DA. The results showed that both the Bmax of 3H-WIN 35,428 binding and the Vmax of 3H-DA reuptake of the patients were significantly lower than those of healthy subjects, while the Kd or Km did not show any change. These findings, while indicating a reduced density of the lymphocyte DAT proteins, provide further support of the role of DA in psychoses and suggest that DA alterations may not be limited to brain structures.

[(3)H]-YM-09151-2 binding sites in human brain postmortem.

The controversial and limited data on the distribution of dopamine (DA) receptors of type 4 (D(4)) in the human brain prompted us to explore their density and pharmacological characteristics in the prefrontal cortex, striatum and hippocampus, through a series of binding assays. Brain samples were taken during autopsy from seven subjects. Tissue homogenates were incubated with increasing concentration of [(3)H]-YM-09151-2, a D(2)-like receptor antagonist, and L-745,870 and/or sulpiride to define the non-specific binding, while PPAP was used to block sigma receptors. The results showed a low density of D(4) receptors in the hippocampus only, with a preponderance of D(2)/D(3) and sigma receptors in the prefrontal cortex and striatum. In conclusion, these findings underline that it is possible to label D(4) receptors by means of [(3)H]-YM-09151-2, provided that D(2), D(3) and sigma receptors are blocked.

Presence and characterization of the dopamine transporter in human resting lymphocytes.

The paucity of information on the presence of the dopamine transporter (DAT) in blood cells, prompted us to explore it in human resting lymphocytes by means of the binding of 3H-WIN 35,428, a compound which is currently considered the most selective ligand for labelling this protein, and by means of the specific reuptake of 3H-dopamine (3H-DA). Lymphocytes were obtained by 15 healthy subjects. The results showed the presence of a specific and saturable binding of 3H-WIN 35,428, which labelled one site only. A specific 3H-DA reuptake was also measured. The pharmacological characterization of both binding and reuptake was overlapping. These findings would indicate that human resting lymphocytes carry the DAT, whose functions in periphery are still unknown.

Presence of D4 dopamine receptors in human prefrontal cortex: a postmortem study.

OBJECTIVE: The aim of our study was to explore the presence and the distribution of D4 dopamine receptors in postmortem human prefrontal cortex, by means of the binding of [3H]YM-09151-2, an antagonist that has equal affinity for D2, D3 and D4 receptors. It was therefore necessary to devise a unique assay method in order to distinguish and detect the D4 component. METHOD: Frontal cortex samples were harvested postmortem, during autopsy sessions, from 5 subjects. In the first assay, tissue homogenates were incubated with increasing concentrations of [3H]YM-09151-2, whereas L-745870, which has a high affinity for D4 and a low affinity for D2/D3 receptors, was used as the displacer. In the second assay, raclopride, which has a high affinity for D2/D3 receptors and a low affinity for D4 receptors, was used to block D2/D3. The L-745870 (500 nM) was added to both assays in order to determine the nonspecific binding. RESULTS: Our experiments revealed the presence of specific and saturable binding of [3H]YM-09151-2. The blockade of D2 and D3 receptors with raclopride ensured that the D4 receptors were labeled. The mean maximum binding capacity was 88 +/- 25 fmol/mg protein, and the dissociation constant was 0.8 +/- 0.4 nM. DISCUSSION AND CONCLUSIONS: Our findings, although not conclusive, suggest that the density of D4 receptors is low in the human prefrontal cortex.

Presence of D4 dopamine receptors in human prefrontal cortex: a postmortem study

OBJECTIVE: The aim of our study was to explore the presence and the distribution of D4 dopamine receptors in postmortem human prefrontal cortex, by means of the binding of [³H]YM-09151-2, an antagonist that has equal affinity for D2, D3 and D4 receptors. It was therefore necessary to devise a unique assay method in order to distinguish and detect the D4 component. METHOD: Frontal cortex samples were harvested postmortem, during autopsy sessions, from 5 subjects. In the first assay, tissue homogenates were incubated with increasing concentrations of [³H]YM-09151-2, whereas L-745870, which has a high affinity for D4 and a low affinity for D2/D3 receptors, was used as the displacer. In the second assay, raclopride, which has a high affinity for D2/D3 receptors and a low affinity for D4 receptors, was used to block D2/D3. The L-745870 (500 nM) was added to both assays in order to determine the nonspecific binding. RESULTS: Our experiments revealed the presence of specific and saturable binding of [³H]YM-09151-2. The blockade of D2 and D3 receptors with raclopride ensured that the D4 receptors were labeled. The mean maximum binding capacity was 88 ± 25 fmol/mg protein, and the dissociation constant was 0.8 ± 0.4 nM. DISCUSSION AND CONCLUSIONS: Our findings, although not conclusive, suggest that the density of D4 receptors is low in the human prefrontal cortex.

OBJETIVO: O objetivo deste estudo foi quantificar a presença e a distribuição de receptores dopaminérgicos do tipo 4 (D4) no córtex cerebral humano em amostras post-mortem através do bloqueio com ³H-YM-09151-2 - um antagonista com afinidade equivalente pelos receptores D2, D3 e D4 - e do desenvolvimento de um método para a detecção específica do componente D4. MÉTODO: Foram obtidas amostras de córtex cerebral de cinco cadáveres. Em um primeiro ensaio, os homogeneizados de tecido cerebral foram incubados em concentrações crescentes de ³H-YM-09151-2, enquanto que o L-745,870, ligante que apresenta grande afinidade pelo receptor D4 e baixa afinidade por D2 e D3, foi utilizado como controle. Em um segundo ensaio, a racloprida, que apresenta alta afinidade por receptores D2 e D3, mas baixa afinidade por D4, foi usada para bloquear D2 e D3. O L-745,870 foi adicionado em ambos os ensaios para determinar o bloqueio não específico. RESULTADOS: Os resultados do experimento demonstraram a presença de um bloqueio específico e saturável com ³H-YM-09151-2. O bloqueio de receptores D2 e D3 com racloprida confirmou que apenas os receptores D4 livres foram avaliados. A Bmax (média ± DP) foi de 88 ± 25 fmol/mg de proteínas, enquanto que a Kd (média ± DP) foi de 0,8 ± 0,4 nM. DISCUSSÃO E CONCLUSÕES: Tais achados, ainda que não definitivamente conclusivos, sugerem a presença de uma baixa densidade de receptores D4 no córtex pré-frontal humano.

Effects of A1 adenosine receptor stimulation on the expression of genes involved in calcium homeostasis.

We investigated whether A(1) adenosine receptor stimulation affects expression of genes involved in calcium homeostasis, including sarcolemmal L-type Ca(2+) channel, Na(+)/Ca(2+) exchanger, sarcoplasmic reticulum (SR) Ca(2+)-ATPase, phospholamban, or ryanodine receptor. Three models of A(1) stimulation were used: i) an acute model, i.e. isolated perfused rat hearts treated for 120 min with 15 nM R-phenylisopropyladenosine (R-PIA), an A(1) receptor agonist; ii) a subacute model, i.e. rats treated with 1.5 mg/kg R-PIA e.v. and sacrificed after 24 h; iii) a transgenic model, i.e. mice overexpressing A(1) adenosine receptors. In all models gene expression was determined by RT-PCR, and oxalate-supported Ca(2+) uptake, representing SR Ca(2+) uptake, was measured in the crude homogenate. Significant increase in the expression of the phospholamban gene was observed in each model of A(1) stimulation, while the expression of the other four genes was not significantly modified. In the acute model, SR Ca(2+) uptake was unaffected, however in the subacute and transgenic models uptake rate was significantly reduced. In parallel experiments, hearts obtained from the subacute model demonstrated a significant reduction in irreversible tissue injury from 30 min of ischemia and 120 min of reperfusion. Increased resistance to ischemia has already been reported also in our transgenic model. In conclusion, A(1) adenosine receptor stimulation up-regulates phospholamban gene expression, which leads within 24 h to a reduced rate of SR Ca(2+) uptake. Changes in Ca(2+) homeostasis might contribute to the delayed cardioprotective effect of adenosine.

Decreased density of peripheral benzodiazepine receptors in psychiatric patients after a suicide attempt.

To date, two main types of benzodiazepine (BDZ) receptors have been identified: one of these is the so-called central receptor which is found mainly in the cortex, limbic areas and cerebellum, and the other is known as the peripheral receptor, which is found in the kidneys, lungs, ovaries, testes, adrenal glands and blood cells, but is present also in the central nervous system (CNS), in particular in glial cells. Although for some time the peripheral BDZ receptor has been considered an acceptor site with no pharmacological activity, recent data have suggested that it may be involved in a variety of actions, such as the response to stress. The presence of these receptors in blood platelets, which are considered a reliable, peripheral mirror of the same structures located in the SNC, prompted us to evaluate them in a group of psychiatric patients after a suicide attempt, as compared with healthy control subjects, by means of the specific binding of 3H-PK 11195. Suicide, with no doubt, may be considered one of the most stressful situations occurring to humans. The results showed the presence of a significant decrease in the density of 3H-PK 11195 binding sites in the patients, as compared with healthy control subjects. This finding may represent a non-specific indicator of a condition of stress, since peripheral BDZ receptors are modulated by stress and hormones, or it may result more from an abnormal metabolism of steroid substances which could play a pivotal role in the development of vulnerability towards suicide.

Changes in peripheral benzodiazepine receptors in patients with bipolar disorder.

Peripheral benzodiazepine (BDZ) receptors were investigated by means of the binding of the specific ligand (3)H-PK 11195 to platelet membranes in patients suffering from bipolar disorder and in healthy controls. The results showed that the density (Bmax) of peripheral BDZ receptors was significantly higher in patients than in control subjects, with no change in the dissociation constant. No correlation with demographic or clinical features was observed. These findings would suggest that alterations of peripheral BDZ receptors are present in patients suffering from bipolar disorder, but it is premature to conclude whether they may be related to the pathophysiology of the disorder, or are secondary to changes occurring in other systems, such as those regulating the stress response.

Platelet alpha2-adrenoreceptors in obsessive-compulsive disorder.

The need for new therapeutic targets in obsessive-compulsive disorder (OCD) prompted us to investigate the putative involvement of the norepinephrine system by means of platelet alpha(2)-adrenoreceptors in a group of 20 OCD patients and healthy control subjects, matched for sex and age. Platelet membranes were prepared according to standard protocols, and the alpha(2)-adrenoreceptors were measured by means of the specific binding of [(3)H]rauwolscine, a highly selective antagonist for this receptor subtype. The results, which showed no difference between patients and controls in the binding parameters of [(3)H]rauwolscine, suggest that the role of alpha(2)-adrenoreceptors, as reflected by the platelet model, is quite limited in OCD and may, perhaps, be restricted purely to some symptoms or dimensions such as motricity, as suggested by the higher density of alpha(2)-adrenoreceptors found in patients concomitantly affected by motor tics.

Normal and obsessional jealousy: a study of a population of young adults.

BACKGROUND: Jealousy is a heterogenous emotion ranging from normality to pathology. Several problems still exist in the distinction between normal and pathological jealousy. AIM OF THE STUDY: With the present study, we aimed to contribute to the definition of the boundary between obsessional and normal jealousy by means of a specific self-report questionnaire developed by us. METHOD: The questionnaire called "Questionnaire on the Affective Relationships" (QAR) and consisting of 30 items, was administered to 400 university students of both sexes and to 14 outpatients affected by obsessive-compulsive disorder (OCD) whose main obsession was jealousy. The total scores and single items were analysed and compared. RESULTS: Two hundred and forty-five, approximately 61% of the questionnaires, were returned. The statistical analyses showed that patients with OCD had higher total scores than healthy subjects; in addition, it was possible to identify an intermediate group of subjects, corresponding to 10% of the total, who were concerned by jealousy thoughts around the partner, but at a lower degree than patients, and that we called "healthy jealous subjects" because they had no other psychopathological trait. Significant differences were also observed for single items in the three groups. CONCLUSIONS: Our study showed that 10% of a population of university students, albeit normal, have jealousy thoughts around the partner, as emerged by the specific questionnaire developed by us. This instrument permitted to clearly distinguish these subjects from patients with OCD and healthy subjects with no jealousy concern.

Decreased lymphocyte 3H-paroxetine binding in obsessive-compulsive disorder.

The pathophysiology of obsessive-compulsive disorder (OCD) is mainly focused on the serotonin (5-HT) system and transporter. The presence of this structure in blood lymphocytes prompted us to investigate it, by means of the specific binding of (3)H-paroxetine ((3)H-PAR), in a group of drug-free OCD patients as compared with healthy control subjects matched for sex and age. Lymphocyte membranes and (3)H-PAR binding were carried out according to standard protocols. The results showed that the patients had a statistically significant lower density of (3)H-PAR-binding sites than the control subjects. On one hand, this finding confirms previous data of an abnormal platelet 5-HT transporter in OCD, on the other it provides the possibility to explore the regulation of this structure in this and other disorders, since lymphocytes are nucleate cells.

Jealousy and subthreshold psychopathology: a serotonergic link.

BACKGROUND: A few studies suggest that different neurotransmitters may play a role in the expression of jealousy. Our study aimed to explore the serotonergic system by means of the specific binding of (3)H-paroxetine ((3)H-Par) to platelet membranes of healthy subjects with and without excessive jealousy concerns, according to a specific self-administered questionnaire [Questionnaire for affective relationships (QAR)]. SUBJECTS AND METHODS: The study sample includes 21 subjects concerned by jealousy thoughts and 21 control subjects without jealousy concerns, as shown by their QAR scores. Subjects of the first group were administered a battery of self-report instruments designed to detect the presence of typical, atypical and subthreshold psychopathology. Platelet membranes and (3)H-Par binding were carried out according to standardized protocols. RESULTS: Subjects with excessive jealousy concerns had a reduced density of (3)H-Par binding as compared with control subjects without jealousy concerns and had one or more psychiatric spectrum conditions. CONCLUSIONS: Our findings suggest that excessive jealousy is associated with various forms of psychopathology and may be underlain by alteration of the serotonergic system, as reflected by the lower density of the platelet serotonin transporter.

Decreased inhibitory activity of PKC in OCD patients after six months of treatment.

We investigated 5-HT reuptake and protein kinase of type C (PKC) activation in platelets of 14 OCD patients at baseline and after six months of treatment with different serotonin (5-HT) reuptake inhibitors (SRIs). The results showed that all SRIs provoked a significant increase in both the maximal velocity (V(max)) and the Michaelis-Menten constant (K(m)) of 5-HT reuptake, as compared with baseline values. The activation of PKC by means of 4-beta-12-tetradecanoylphorbol 13-acetate provoked a significant decrease in V(max) values, but the effect was not as evident as at baseline. These findings could indicate that, in OCD patients, SRIs increase the rate of reuptake and decrease the inhibitory effect of PKC and that the two phenomena may be linked, the first perhaps depending upon the second.