Joanna Listos

Effects of sildenafil treatment on the development of tolerance to diazepam-induced motor impairment and sedation in mice

We studied the effects of sildenafil, a selective inhibitor of PDE5, on the development and the expression of tolerance to diazepam (DZ)-induced motor impairment and sedation in mice. DZ-induced motor incoordination was assessed by the rotarod and chimney tests, and DZ-induced sedation was examined using a photocell apparatus. Sildenafil treatment enhanced the development of tolerance to the motor impairing effects, but not to the sedative effects, of DZ. Sildenafil treatment did not affect the expression of tolerance to DZ-induced motor impairment and sedation in mice. Our results suggest that sildenafil treatment, at least in part, affects the development of DZ tolerance.

Adenosine receptor agonists attenuate the development of diazepam withdrawal-induced sensitization in mice

In the present study, the effects of adenosine agonists on the development of sensitization to withdrawal signs precipitated after sporadic treatment with diazepam, in mice, were investigated. To obtain the sensitization, the animals were divided into groups: continuously and sporadically treated with diazepam (15.0 mg/kg, s.c.). The adenosine receptor agonists (CPA, CGS 21,680 and NECA) were administered in sporadically diazepam treated mice during two diazepam-free periods. Concomitant administration of pentetrazole (55.0 mg/kg, s.c.) with flumazenil (5.0 mg/kg, i.p.) after the last injection of diazepam or vehicle, induced the withdrawal signs, such as clonic seizures, tonic convulsion and death episodes. The major finding of our experiments is attenuation of withdrawal signs in sensitized mice, inducing by all adenosine agonists. Only higher dose of CPA produced significantly decreased the number of withdrawal incidents, while both used doses of CGS 21,680 and NECA produced more clear effects. These results support the hypothesis that adenosinergic system is involved in the mechanisms of sensitization to the benzodiazepine withdrawal signs, and adenosine A(2A) receptors play more important role in that process.

Effects of NOS inhibitors on the benzodiazepines-induced memory impairment of mice in the modified elevated plus-maze task.

The aim of the present study was to examine the effects of nitric oxide synthase (NOS) inhibitors on responses, elicited by benzodiazepines (BZs) in a modified elevated plus-maze task in mice. It was shown that acute doses of diazepam (DZ; 1 and 2 mg/kg) and flunitrazepam (FNZ; 0.05, 0.1 and 0.2 mg/kg) significantly increased the time of transfer latency (TL2) in a retention trial, thus confirming memory impairing effects of BZs. l-NAME (N(G)-nitro-l-arginine methyl ester; 200 mg/kg), a non-selective inhibitor of NOS, and 7-NI (7-nitroindazole; 40 mg/kg), a selective inhibitor of NOS, further intensified DZ-induced memory impairment. On the other hand, L-NAME (50, 100 and 200 mg/kg) and 7-NI (10, 20 and 40 mg/kg) prevented FNZ-induced memory compromising process. The results of this study indicated that suppressed NO synthesis enhanced DZ-induced but prevented FNZ-induced memory impairment. Taken together, these findings could suggest NO involvement in BZs-induced impairment of memory processes. The precise mechanism of these controversial effects, however, remains elusive.

The effect of perinatal lead exposure on dopamine receptor D2 expression in morphine dependent rats.

The aim of this study was to investigate the behavioral and molecular effects of pre- and postnatal lead (Pb) exposure on the expression of morphine withdrawal and tolerance in adult rats. Rats were orally treated with 0.1% (1000ppm) lead acetate from conception, through gestation, up to postnatal day (PND) 28. Subsequently, behavioral experiments were conducted on adult (PND 60) male rats. To assess behavioral effects of morphine dependence in Pb-exposed rats two experimental models were used: naloxone-precipitated withdrawal signs and the assessment of morphine tolerance to antinociceptive effect in the tail-immersion test. Morphine withdrawal and tolerance were more expressed in Pb-exposed morphine administered rats than in morphine administered rats. In the case of morphine withdrawal signs the analysis of protein (Western blotting) and mRNA (RT PCR) expression revealed significantly higher dopamine D2 receptor (D2R) expression in prefrontal cortex, but not in striatum and hippocampus, in Pb-exposed morphine administered rats than in morphine administered rats. Differently, in the case of morphine tolerance the significant upregulation of D2R protein and mRNA expression in hippocampus, but not in prefrontal cortex or striatum, was demonstrated in Pb-exposed and morphine administered rats in comparison with morphine administered. These findings suggest that in morphine withdrawal and tolerant rats the perinatal Pb-exposure can affect D2R expression in brain region-specific manner. Immunohistochemical assessment of D2R expression in hippocampus showed translocation of D2R from membrane-cytoplasm in control rats to nucleus in morphine administered rats. Perinatal Pb-exposure did not induce the changes in the localization of D2R irrespective of morphine effect.

Effects of perinatal exposure to lead (Pb) on purine receptor expression in the brain and gliosis in rats tolerant to morphine analgesia.

The aim of the present study was to investigate the molecular effects of perinatal exposure to lead (Pb) on protein and mRNA expression of purine receptors: P2X4, P2X7, adenosine receptor A1; and astrocytes (GFAP mRNA expression) and on microglia activation (Iba1 mRNA expression) in several structures of the mesolimbic system (striatum, hippocampus, prefrontal cortex) in rats expressing tolerance to the antinociceptive effect of morphine. Rat mothers were orally treated with 0.1% lead acetate from conception, through gestation, and postnatally, as well as to offspring up to day (PND) 28; subsequently molecular studies were conducted on adult (PND 60) male rats. Morphine tolerance developed more strongly in rats perinatally exposed to Pb. The analysis revealed a significant up-regulation of protein and mRNA P2X4 receptor expression in the striatum and prefrontal cortex but not in the hippocampus; P2X7 protein and mRNA receptor expression in the striatum and hippocampus, but not in the prefrontal cortex; A1 protein receptor expression in all investigated structures and A1 mRNA expression in the striatum and hippocampus; Iba1 mRNA expression in the striatum and hippocampus; and GFAP mRNA expression in the striatum and prefrontal cortex. Immunohistochemical analysis has also revealed significant alterations. Strong expressions of P2X4, P2X7, A1 receptors, astrocytes and microglia activation were observed in the hippocampus in Pb and/or morphine treated rats. The higher expression of purine receptors and glial cell activation are important markers of neuroinflammatory processes. Therefore, we conclude that Pb-induced neuroinflammation may be responsible for the intensification of morphine tolerance in the Pb-treated rats. Additionally, the dysregulation of A1 adenosine receptors, mainly in the hippocampus, may also be involved in the intensification of morphine tolerance in Pb-treated rats. Our study demonstrates the significant participation of environmental factors in addictive process; additionally, it shows the necessity of modification of addictive disorder with neuroprotective agents.

Influence of a low dose of silver nanoparticles on cerebral myelin and behavior of adult rats.

Nanoscale particles have large surface to volume ratio that significantly enhances their chemical and biological reactivity. Although general toxicity of nano silver (nanoAg) has been intensively studied in both in vitro and in vivo models, its neurotoxic effects are poorly known, especially those of low-dose exposure. In the present study we assess whether oral administration of nanoAg influences behavior of exposed rats and induces changes in cerebral myelin. We examine the effect of prolonged exposure of adult rats to small (10nm) citrate-stabilized nanoAg particles at a low dose of 0.2mg/kg b.w. (as opposed to the ionic silver) in a comprehensive behavioral analysis. Myelin ultrastructure and the expression of myelin-specific proteins are also investigated. The present study reveals slight differences with respect to behavioral effects of Ag(+)- but not nanoAg-treated rats. A weak depressive effect and hyperalgesia were observed after Ag(+) exposure whereas administration of nanoAg was found to specifically increase body weight and body temperature of animals. Both nanoAg and Ag(+) induce morphological disturbances in myelin sheaths and alter the expression of myelin-specific proteins CNP, MAG and MOG. These results suggest that the CNS may be a target of low-level toxicity of nanoAg.

Effects of NMDA antagonists on the development and expression of tolerance to diazepam-induced motor impairment in mice

The goal of the study was to investigate the effects of ketamine and memantine on the development and expression of tolerance to diazepam (DZ)-induced motor impairment in mice. DZ-induced motor incoordination was assessed by the rotarod and chimney tests. It was found that (a) ketamine, at the dose of 5mg/kg (but not 2.5mg/kg), decreased the expression, but not the development, of tolerance to the motor impairing effects of DZ, (b) memantine, at the doses of 5 and 10mg/kg decreased both the development and expression of DZ tolerance in the rotarod test (also in the chimney test but at the higher dose of 10mg/kg) and (c) ketamine and memantine alone had no effect, either in the rotarod or the chimney test in mice. Those findings provided behavioral evidence that the glutamatergic system could contribute an important role in the development and/or expression of tolerance to DZ in mice.

Adenosinergic system is involved in development of diazepam tolerance in mice

In the present study the effect of adenosinergic system on the development of diazepam tolerance to motor disturbances in mice was investigated. Diazepam tolerance was obtained by administration of diazepam at a dose of 5.0 mg/kg, s.c. for ten consecutive days. On the 1st and the 10th day of the experiment motor impairments were measured in two behavioural tests: rota-rod and chimney test. We showed that acute diazepam injection produced significant motor impairments in mice and that effect was decreased by repeated diazepam treatment, confirming the development of tolerance to the motor impairing effect of diazepam. We demonstrated that adenosine A(1) and/or A(2A) receptor agonists: CPA (0.025 and 0.05 mg/kg, i.p.), CGS 21680 (0.1 and 0.2 mg/kg, i.p.), NECA (0.005 and 0.01 mg/kg, i.p.) pretreatment with diazepam were able to attenuate the development of diazepam tolerance and adenosine receptor antagonists: DPCPX (1.0 and 3.0 mg/kg, i.p.), DMPX (3.0 and 6.0 mg/kg, i.p.) and caffeine (10.0 and 20.0mg/kg, i.p.) induced the opposite effect. The most apparent effects were obtained by non-selective agonist (NECA) and antagonist (caffeine) of adenosine receptors. We conclude that adenosinergic system plays an important role in mechanisms underlying the development of benzodiazepine tolerance.

Divergent effects of l-arginine-NO pathway modulators on diazepam and flunitrazepam responses in NOR task performance

The goal of the study was an evaluation of the degree, in which nitric oxide (NO) is involved in the benzodiazepines (BZs)-induced recognition memory impairment in rats. The novel object recognition (NOR) test was used to examine recognition memory. The current research focused on the object memory impairing effects of diazepam (DZ; 0.5 and 1mg/kg, sc) and flunitrazepam (FNZ; 0.1 and 0.2mg/kg; sc) in 1-hour delay periods in rats. It was found that acute ip injection of L-arginine (L-arg; 250 and 500 mg/kg; ip), 5 min before DZ administration (0.5mg/kg, sc) prevented DZ-induced memory deficits. On the other hand, it was also proven that L-arg (125, 250 and 500 mg/kg; ip) did not change the behaviour of rats in the NOR test, following a combined administration with FNZ at a threshold dose (0.05 mg/kg; sc). It was also found that 7-nitroindazole (7-NI; 10, 20 and 40 mg/kg; ip) induced amnesic effects in DZ in rats, submitted to the NOR test, following a combined administration of 7-NI with a threshold dose of DZ (0.25mg/kg; sc). However, following a combined administration of 7-NI (10, 20 and 40 mg/kg; ip) with FNZ (0.1mg/kg; sc), it was observed that 7-NI inhibited the amnesic effects of FNZ on rats in the NOR test. Those findings led us to hypothesize that NO synthesis suppression may induce amnesic effects of DZ, while preventing FNZ memory impairment in rats, submitted to NOR tasks.

Effects of chronic flunitrazepam treatment schedule on therapy-induced sedation and motor impairment in mice

BACKGROUND The aim of the present study was to examine whether different treatment schedules could be associated with tolerance development to the ataxic and sedative effects of flunitrazepam in mice. METHODS Effects of repeated flunitrazepam administration were studied in the rotarod and the chimney test for motor coordination and in a photocell apparatus for locomotor activity in mice. Flunitrazepam doses varied in particular types of injections or in different experiment duration periods. RESULTS Repeated flunitrazepam administration (1 mg/kg, sc and 2 mg/kg, ip) for 8 consecutive days induced tolerance to the motor impairing effects of flunitrazepam in mice, both in the rotarod and the chimney test. In turn, no tolerance developed to sedative flunitrazepam effects, regarding either dose level, injection type or treatment duration. CONCLUSIONS Those findings confirmed the previous observations that tolerance to benzodiazepines was not simultaneous for each pharmacological property of the drugs. Interestingly enough, an acute dose of flunitrazepam (1 mg/kg, sc) in our study enhanced locomotor activity of mice.