Effect of antidepressant drugs in mice lacking the norepinephrine transporter.

One of the main theories concerning the mechanism of action of antidepressant drugs (ADs) is based on the notion that the neurochemical background of depression involves an impairment of central noradrenergic transmission with a concomitant decrease of the norepinephrine (NE) in the synaptic gap. Many ADs increase synaptic NE availability by inhibition of the reuptake of NE. Using mice lacking NE transporter (NET-/-) we examined their baseline phenotype as well as the response in the forced swim test (FST) and in the tail suspension test (TST) upon treatment with ADs that display different pharmacological profiles. In both tests, the NET-/- mice behaved like wild-type (WT) mice acutely treated with ADs. Autoradiographic studies showed decreased binding of the beta-adrenergic ligand [3H]CGP12177 in the cerebral cortex of NET-/- mice, indicating the changes at the level of beta-adrenergic receptors similar to those obtained with ADs treatment. The binding of [3H]prazosin to alpha1-adrenergic receptors in the cerebral cortex of NET-/- mice was also decreased, most probably as an adaptive response to the sustained elevation of extracellular NE levels observed in these mice. A pronounced NET knockout-induced shortening of the immobility time in the TST (by ca 50%) compared to WT mice was not reduced any further by NET-inhibiting ADs such as reboxetine, desipramine, and imipramine. Citalopram, which is devoid of affinity for the NET, exerted a significant reduction of immobility time in the NET-/- mice. In the FST, reboxetine, desipramine, imipramine, and citalopram administered acutely did not reduce any further the immobility time shortened by NET knockout itself (ca 25%); however, antidepressant-like action of repeatedly (7 days) administered desipramine was observed in NET-/- mice, indicating that the chronic presence of this drug may also affect other neurochemical targets involved in the behavioral reactions monitored by this test. From the present study, it may be concluded that mice lacking the NET may represent a good model of some aspects of depression-resistant behavior, paralleled with alterations in the expression of adrenergic receptors, which result as an adaptation to elevated levels of extracellular NE.

Effects of tramadol on alpha2-adrenergic receptors in the rat brain.

In recent years, it has been postulated that tramadol, used mainly for the treatment of moderate to severe pain, might display a potential as an antidepressant drug. The present study investigated the effects of acute and repeated tramadol administration on the binding of [3H]RX 821002, a selective alpha2-adrenergic receptor ligand, in the rat brain. Male Wistar rats were used. Tramadol (20 mg/kg, i.p.) administered acutely (single dose), at 24 h after dosing, induced a significant decrease in the alpha2-adrenergic receptors in all brain regions studied. The most pronounced effects were observed in all subregions of the olfactory system, nucleus accumbens and septum, thalamus, hypothalamus, amygdala, and cerebral cortex. Repeated treatment with tramadol (20 mg/kg, i.p., once daily for 21 days) also induced statistically significant downregulation of [3H]RX 821002 binding sites in the rat brain. However, the effect--although statistically significant--was less pronounced than in the group treated acutely with the drug. Since drugs such as mianserin and mirtazapine are potent antagonists of central alpha2-adrenergic receptors and are effective antidepressants, it is tempting to suggest that, in addition to other alterations induced by tramadol, downregulation of these receptors may represent a potential antidepressant efficacy. On the other hand, one should be careful to avoid the treatment of chronic pain with tramadol in patients already receiving antidepressant drugs. Tramadol-induced downregulation of alpha2-adrenergic receptors--when combined with ongoing antidepressant therapy with drugs, which themselves inhibit serotonin reuptake or are antagonists of alpha2-adrenergic receptors--might cause threatening complications.

Long-term exposure of rats to tramadol alters brain dopamine and alpha 1-adrenoceptor function that may be related to antidepressant potency.

The aim of the present study was to determine whether tramadol, which has a potential antidepressant efficacy, evokes, when administered repeatedly, changes similar to the alterations induced by conventional antidepressant drugs. Repeated administration of tramadol (20 mg/kg i.p. for 21 days) enhanced the d-amphetamine-induced locomotor hyperactivity and increased the density of alpha(1)-adrenoceptors in the rat brain cortex, as measured by saturation analysis of [(3)H]prazosin binding. Autoradiographic analysis of [(3)H]7-OH-DPAT and [(3)H]raclopride binding revealed a significant up-regulation of dopamine D2 and D3 receptors in the rat nucleus accumbens upon repeated treatment with tramadol. All the above-mentioned effects induced by repeated administration of tramadol resemble the effects induced by conventional antidepressants. However, tramadol when administered repeatedly did not increase the levels of mRNA encoding for brain-derived neurotrophic factor (BDNF) and its receptor, TrkB. This is what differs tramadol from conventional antidepressants, since neurotrophic effects of these drugs have recently been postulated.

The effect of cocaine sensitization on mouse immunoreactivity.

Recent studies indicate a role of the immune system in the behavioral effects of cocaine in rodents. In the present study, we attempted to find a correlation between the behavioral changes induced by repeated, intermittent administration of cocaine and some immunological consequences of sensitization to cocaine. Male Albino Swiss mice were treated repeatedly (for 5 days) with cocaine (10 or 15 mg/kg, intraperitoneally, ip). On day 9, they received a challenge dose of cocaine (10 or 15 mg/kg). Acute administration of cocaine increased the locomotor activity of mice. In animals treated repeatedly with the higher dose of cocaine, the locomotor hyperactivity induced by a challenge dose of the psychostimulant (15 mg/kg) was ca. twice as high as that after its first administration; in consequence, evidence for behavioral sensitization was obtained. Immune functions were evaluated by measuring the ability of splenocytes to proliferate and to produce cytokines such as interferon-gamma (IFN-gamma), interleukin (IL)-4 and IL-10. Acute cocaine administration significantly decreased proliferation of splenocytes to concanavalin A (Con A) and increased their ability to produce IFN-gamma. Repeated intermittent treatment with cocaine in a dose of 10 mg/kg significantly decreased the thymus weight and the proliferative response of T cells to a suboptimal dose of Con A. Sensitization with the higher dose of cocaine significantly enhanced IFN-gamma production. These data indicate that cocaine sensitization results in the development of a tolerant state to the cocaine-induced suppression of a thymus dependent T-lymphocyte response. It may be suggested that the cocaine sensitization partly depends on the altered balance of cytokine production, e.g. an increase in IFN-gamma production. Since repeated, intermittent use of cocaine by humans leads to psychoses or craving for this drug, our findings also seem to indicate considerable importance of monitoring and correcting immune changes in the therapy of cocaine addiction.

Effects of 5-HT1B receptor ligands microinjected into the ventral tegmental area on the locomotor and sensitizating effects of cocaine in rats.

The present study was aimed at finding out whether 5-HT(1B) receptors located in the ventral tegmental area (VTA) played a role in the locomotor hyperactivity induced by a single dose of cocaine and in the sensitization evoked by repeated exposure to the psychostimulant in rats. Male Wistar rats, implanted bilaterally with cannulae in the VTA, were microinjected with GR 55562 (an antagonist of 5-HT(1B) receptors) or CP 93129 (an agonist of 5-HT(1B) receptors). GR 55562 (0.3-3 microg/side) did not affect locomotor hyperactivity response to a single dose of cocaine (10 mg/kg). CP 93129 in a dose of 1 microg/side (but not lower), which stimulated basal locomotor activity, enhanced the cocaine-induced locomotor hyperactivity. The rats that were treated repeatedly (for 5 days) with cocaine (10 mg/kg) and then challenged with cocaine (10 mg/kg) after 5-day withdrawal period (day 10 of the experiment) showed significantly higher locomotor hyperactivity compared to the effect observed in saline-pretreated and cocaine-challenged rats. GR 55562 (a dose of 3 microg/side, but not lower), administered for 5 days into the VTA just prior to daily cocaine, attenuated cocaine sensitization. When injected for 5 days into the VTA, CP 93129 (0.03-0.1 microg/side) enhanced the development of cocaine sensitization. The enhancing effect of CP 93129 (0.1 microg/side) was blocked by GR 55562 (1 microg/side). To examine the effects of GR 55562 and CP 93129 on the expression of cocaine sensitization, the 5-HT(1B) receptor ligands were given acutely before the challenge dose of cocaine administered on day 10. No change in cocaine sensitization was observed after intra-VTA microinjections of GR 55562 (0.3-3 microg/side) or CP 93129 (0.1-1 microg/side). Our findings suggest that 5-HT(1B) receptors located in the VTA play a permissive role in the development of cocaine sensitization, but are not involved in the locomotor hyperactivity induced by a single dose of cocaine or in the expression of the sensitization to the psychostimulant.

Effects of 5-HT(1B) receptor ligands microinjected into the accumbal shell or core on the sensitization to cocaine in rats.

The present study was designed to find out whether 5-HT(1B) receptors located in subareas of the nucleus accumbens played a role in cocaine sensitization in rats, and whether pharmacological activation of these receptors could modify this drug effect. Male Wistar rats implanted bilaterally with cannulae into the accumbens shell or core were microinjected with GR 55562 (an antagonist of 5-HT(1B) receptors) or CP 93129 (an agonist of 5-HT(1B) receptors). The rats, which were repeatedly (for 5 days) administered with cocaine (10 mg/kg) and then challenged with cocaine (10 mg/kg) after 5-day withdrawal period, showed significantly higher locomotor hyperactivity in comparison with the effect observed in saline-pretreated and cocaine challenged rats. GR 55562 (0.1-10 microg/side), administered for 5 days into the accumbens shell, but not into the core, prior to cocaine dose-dependently attenuated cocaine sensitization. When injected for 5 days into either the accumbens shell or core before cocaine, CP 93129 (0.1-10 microg/side) had no effect on the development of cocaine sensitization. To examine the effects of GR 55562 and CP 93129 on the expression of cocaine sensitization, the drugs were given acutely before a challenge dose of cocaine (10 mg/kg) on day 10. No change in cocaine sensitization was observed after injection of GR 55562 (0.1-10 microg/side) to the either accumbens subregion or after injection of CP 93129 (0.1-10 microg/side) into the core. However, an intra-accumbens shell injection of CP 93129 (10-30 microg/side) increased sensitization to cocaine, and this enhancement was attenuated after local injection of GR 55562 (1 microg/side). Our findings indicate that behavioral sensitization to cocaine may be modulated by 5-HT(1B) receptor ligands in the accumbens shell, but not into the core. They suggest that the inhibition of 5-HT(1B) receptors in the accumbens shell attenuates the development, whereas their pharmacological activation enhances the expression of cocaine sensitization.

Effect of some antipsychotic drugs on immunoreactivity in C57BL/6 mice.

The study examined the effect of some typical and atypical antipsychotic drugs on mouse lymphocyte metabolic and proliferative activity in vitro. The typical antipsychotic drug chlorpromazine (3 x 10(-6), 10(-5) and 10(-4) M), significantly inhibited 3H-thymidine incorporation into C57BL/6 mouse spleen cells stimulated by concanavalin A (Con A), lipopolysaccharide (LPS) or pokeweed mitogen (PWM). Chlorpromazine at concentrations of 10(-5) and 10(-4) M also suppressed the metabolic activity of splenocytes after Con A stimulation. The atypical antipsychotic agent clozapine (10(-4) and 10(-5) M) decreased the proliferative activity of splenocytes after LPS stimulation, but its inhibitory effect after Con A was observed only at higher concentrations. On the other hand, clozapine did not affect the metabolic activity of splenocytes. Sulpiride, a selective dopamine D2 antagonist, at concentrations ranging from 10(-8) to 10(-4) M had no inhibitory effect on the proliferative or metabolic activity of the tested cells. The obtained results indicate that of the three antipsychotic drugs studied, chlorpromazine shows the most potent immunosuppressive effect, clozapine produces a moderate effect and sulpiride is totally inactive. These findings suggest that the choice of antipsychotic drugs should also depend on disturbance of immune system activity, in particular, those occurring in the several forms of psychosis.