Rita C. Carvalho

Role of hippocampal oxidative stress in memory deficits induced by sleep deprivation in mice.

Numerous animal and clinical studies have described memory deficits following sleep deprivation. There is also evidence that the absence of sleep increases brain oxidative stress. The present study investigates the role of hippocampal oxidative stress in memory deficits induced by sleep deprivation in mice. Mice were sleep deprived for 72 h by the multiple platform method-groups of 4-6 animals were placed in water tanks, containing 12 platforms (3 cm in diameter) surrounded by water up to 1 cm beneath the surface. Mice kept in their home cage or placed onto larger platforms were used as control groups. The results showed that hippocampal oxidized/reduced glutathione ratio as well as lipid peroxidation of sleep-deprived mice was significantly increased compared to control groups. The same procedure of sleep deprivation led to a passive avoidance retention deficit. Both passive avoidance retention deficit and increased hippocampal lipid peroxidation were prevented by repeated treatment (15 consecutive days, i.p.) with the antioxidant agents melatonin (5 mg/kg), N-tert-butyl-alpha-phenylnitrone (200 mg/kg) or vitamin E (40 mg/kg). The results indicate an important role of hippocampal oxidative stress in passive avoidance memory deficits induced by sleep deprivation in mice.

Important role of striatal catalase in aging- and reserpine-induced oral dyskinesia.

Tardive dyskinesia, the most serious iatrogenic movement disorder, has been tentatively associated with nigrostriatal dopaminergic supersensitivity and with oxidative stress. It is also suggested that long-term neuroleptic treatment does not cause oral dyskinesia (OD), but interacts with some substrate of brain aging, resulting in the premature emergence of OD, that can occur spontaneously with aging. In order to investigate a possible role of nigrostriatal dopaminergic supersensitivity and of oxidative stress in aging- and reserpine-induced OD, the stereotyped behavior induced by dopaminergic agonists, a functional index of dopaminergic striatal activity, as well as the striatal antioxidant enzymes glutathione peroxidase and catalase were assessed. We demonstrate that, opposite to normotensive Wistar rats (NWR), spontaneously hypertensive rats (SHR) do not develop aging- or reserpine-OD. There were no differences between NWR and SHR in stereotyped behavior or in striatal glutathione peroxidase activity. Adult and old SHR presented higher striatal catalase activity relative to NWR, and aging increased it only in SHR. The catalase inhibitor aminotriazole reverted the absence of aging- and reserpine-induced OD in SHR. Our results suggest an important role of striatal catalase in the development of reserpine- and aging-induced OD.

Behavioral characterization of morphine effects on motor activity in mice

A biphasic effect of morphine on locomotion has been extensively described. Nevertheless, the effects of this opioid on other behavioral parameters have been overlooked. The aim of the present study was to verify the effects of different doses of morphine on motor behaviors observed in an open-field. Adult female mice were injected with saline or morphine (10, 15 and 20 mg/kg, i.p.) and observed in an open-field for quantification of locomotor and rearing frequencies as well as duration of immobility and grooming. The lowest dose of morphine decreased locomotion (and increased immobility duration) while the highest dose increased it. All doses tested decreased rearing and grooming. Thus, the effects of morphine on locomotion do not parallel to its effects on rearing and grooming. Our results indicate that locomotion not always reflects the effect of drugs on motor activity, which can be better investigated when other behavioral parameters are concomitantly taken into account.

Beneficial effects of vitamin C and vitamin E on reserpine-induced oral dyskinesia in rats: critical role of striatal catalase activity.

Oral dyskinesias are implicated in a series of neuropathologies and have been associated to an increase in oxidative stress. Several antioxidants, including vitamin E, decrease reserpine-induced oral dyskinesia (OD) in rodents and we have described a protective role of striatal catalase against the development of OD. The aim of this study was to verify the effects of vitamin C alone or in combination with vitamin E on reserpine-induced OD as well as to determine a possible role of catalase in the antidyskinetic property of these vitamins. Different doses of vitamin C attenuated reserpine-induced increase in OD. A similar treatment with an effective dose of vitamin C concomitant to an effective dose of vitamin E potentiated the antidyskinetic effect of both vitamins when administered alone. The administration of these vitamins alone produced an increase in striatal catalase activity that likewise was potentiated by their combined administration. In addition, the antidyskinetic property of vitamin E and vitamin C was abolished by a concomitant treatment with the catalase inhibitor aminotriazole. These results indicate a beneficial effect of these vitamins and reinforce the critical role of striatal catalase against the development of oral dyskinesias.

Concomitant development of oral dyskinesia and memory deficits in reserpine-treated male and female mice

It has been suggested that reserpine-induced oral dyskinesia in rats may provide a new animal model of tardive dyskinesia. Both cognitive deficits and gender have been associated with the development of tardive dyskinesia. The aim of the present study was to investigate the effects of reserpine administration on the development of orofacial dyskinesia and on plus-maze discriminative avoidance task (DAT-an animal model of associative learning) in male and female mice. Male and female mice received 1.0 mg/kg reserpine or saline subcutaneously on day 1. On days 3, 6 and 8, the frequency of vacuous chewing movements (VCM) was quantified. On day 6, the DAT conditioning was performed, in a modified elevated plus-maze. In one of the enclosed arms, the animals received aversive stimulation (light and noise). On day 8, a test session was performed and the time spent by the animals in each of the enclosed arms was recorded. Our results showed that reserpine-treated male and female mice presented significantly higher VCM when compared with respective control groups in all observation days. On day 6, reserpine-treated female mice presented significantly higher VCM when compared with male mice injected with this drug. The DAT test performed on day 8 showed that the time spent in the aversive arm by saline-treated mice was significantly lower than the time spent in the non-aversive arm. This difference was not observed for reserpine-treated mice. Our results demonstrate the development of reserpine-induced oral dyskinesia in both male and female mice. While this oral dyskinesia is accompanied by a cognitive deficit in both genders, female mice tended to have more severe oral dyskinesia. It is suggested that reserpine-induced oral dyskinesia may provide a quick, simple and efficient mouse model of tardive dyskinesia.

The importance of housing conditions on behavioral sensitization and tolerance to ethanol

The differential outcomes of social isolation and crowding environment on the effects of single or repeated administration of ethanol on open-field behavior were examined in female mice. Whereas housing conditions did not alter the increase in locomotor activity induced by ethanol single administration, behavioral sensitization (a progressive increase of a drug effect following repeated drug administration) to the locomotor activating effect of ethanol was significantly greater in crowded mice as compared to isolated and control groups. Single administration of ethanol significantly decreased rearing frequency and increased immobility duration, there being tolerance to these ethanol behavior effects after repeated treatment. Social isolation attenuated the increase in immobility behavior induced by single administration of ethanol and potentiated the tolerance of ethanol-induced rearing decrease, verified after repeated treatment. These results point out that both sensitization and tolerance to the behavioral effects of ethanol can be critically influenced by housing conditions.

Effects of reserpine on the plus-maze discriminative avoidance task: dissociation between memory and motor impairments.

We investigated the effects of reserpine (0.1-0.5 mg/kg) on the performance of mice in the plus-maze discriminative avoidance task (DAVT), which simultaneously evaluates memory and motor activity. All doses induced memory impairment (increased aversive arm time) but only 0.5 mg/kg reserpine decreased locomotion (entries in enclosed arms). The results suggest that the DAVT evaluation in reserpine-treated mice can be a useful model for studying cognitive deficits accompanied by motor impairments.

Haloperidol (but not ziprasidone) withdrawal potentiates sensitization to the hyperlocomotor effect of cocaine in mice.

One important contributing factor in the high prevalence of drug abuse disorders seen among schizophrenic patients seems to be related to chronic treatment with typical neuroleptics. We have previously demonstrated that withdrawal from long-term treatment with the typical neuroleptic haloperidol, but not the atypical neuroleptic ziprasidone, potentiated the hyperlocomotor effect induced by a single cocaine injection and cocaine-induced conditioned place preference in mice. In the present study we investigated whether withdrawal from long-term treatment with these same neuroleptics would also modify cocaine-induced hyperlocomotion sensitization, which has been proposed as an animal model for the intensification of drug craving in cocaine addiction. Swiss male mice were i.p. treated with haloperidol (1.0 mg/kg) or ziprasidone (4.0 mg/kg) for 15 days. Twenty-four hours after the last injection, animals received an i.p. injection of cocaine (10 mg/kg) for 5 consecutive days, being placed after each injection in the open-field apparatus in order to perform a drug-environment conditioning. Seven days after the last drug-environment conditioning procedure, the animals were challenged with an i.p. injection of cocaine (10 mg/kg), placed in the open-field apparatus and had their locomotor activity quantified. Withdrawal from haloperidol (but not ziprasidone) potentiated cocaine-induced behavioral sensitization. These results are suggested to be a consequence of the development of the dopaminergic supersensitivity phenomenon by long-term treatment with the typical compound. Our findings provide additional support for the use of atypical agents like ziprasidone in the treatment of schizophrenic patients with comorbid substance abuse disorder.

Antidyskinetic effects of risperidone on animal models of tardive dyskinesia in mice

The effects of risperidone, an atypical neuroleptic, were investigated on two animal models of tardive dyskinesia (TD). The repeated administration of reserpine (1.0mg/kg) or haloperidol (2.0mg/kg) induces orofacial movements in mice, which are very similar to those observed in humans presenting TD. The effects of acute or repeated treatment with several doses of risperidone (0.1; 0.5; 2.0 or 4.0) on the expression and development of orofacial movements in reserpine- and haloperidol-treated male mice were investigated. The results showed that risperidone per se did not induce the development of orofacial movements. In addition, this drug was able to attenuate the expression and the development of reserpine-as well as haloperidol-induced orofacial movements. These results are in line with several clinical studies that suggest not only a lower incidence of TD in schizophrenic patients treated with risperidone, but also an antidyskinetic effect of this drug in patients previously treated with classical neuroleptics.

Long-term haloperidol treatment (but not risperidone) enhances addiction-related behaviors in mice: role of dopamine D2 receptors.

The high prevalence of psychostimulant abuse observed in schizophrenic patients may be related to the development of mesolimbic dopaminergic supersensitivity (MDS) or nigrostriatal dopaminergic supersensitivity (NDS) in response to the chronic blockade of dopamine receptors produced by typical neuroleptic treatment. We compared the effects of withdrawal from long‐term administration of the typical neuroleptic haloperidol (Hal) and/or the atypical agent risperidone (Ris) on MDS and NDS, behaviorally evaluated by amphetamine‐induced locomotor stimulation (AILS) and apomorphine‐induced stereotypy (AIS) in mice, respectively. We further evaluated the duration of MDS and investigated the specific role of dopamine D2 receptors in this phenomenon by administering the D2 agonist quinpirole (Quin) to mice withdrawn from long‐term treatment with these neuroleptics. Withdrawal (48 hours) from long‐term (20 days) Hal (0.5 mg/kg i.p.) (but not 0.5 mg/kg Ris i.p.) treatment potentiated both AILS and AIS. Ris co‐administration abolished the potentiation of AILS and AIS observed in Hal‐withdrawn mice. Ten days after withdrawal from long‐term treatment with Hal (but not with Ris or Ris + Hal), a potentiation in AILS was still observed. Only Hal‐withdrawn mice presented an attenuation of locomotor inhibition produced by Quin. Our data suggest that the atypical neuroleptic Ris has a pharmacological property that counteracts the compensatory MDS and NDS developed in response to the chronic blockade of dopamine receptors imposed by Ris itself or by typical neuroleptics such as Hal. They also indicate that MDS may be long lasting and suggest that an upregulation of dopamine D2 receptors in response to long‐term treatment with the typical neuroleptic is involved in this phenomenon.