La intoxicación con cobre disminuye la sobrevida e induce alteraciones neurológicas en Drosophila melanogaster / Copper intoxication decreases lifespan and induces neurologic alterations in Drosophila melanogaster

La enfermedad de Wilson, es un trastorno hereditario autosómico recesivo causado por mutaciones del gen de la trifosfatasa de adenosina (ATP7B). Dicha mutación ocasiona intoxicación con cobre, generando manifestaciones clínicas por los efectos tóxicos del metal, principalmente a nivel del hígado y el encéfalo. Recientemente se han desarrollado modelos genéticos de la enfermedad para su estudio clínico. Sin embargo, la utilidad de los mismos es limitada por el hecho de que en tales modelos no se observan manifestaciones neurológicas. El presente estudio tuvo como objetivo desarrollar un modelo de la enfermedad de Wilson en Drosophila melanogaster. Inicialmente se evaluó el efecto de la suplementación con concentraciones de 31 µM y 47 µM de cobre en la sobrevida. Posteriormente se realizaron estudios de conducta para determinar si existían alteraciones en el desempeño motor asociadas al tratamiento con la dosis de 47 µM de cobre. Los resultados obtenidos sugieren que el tratamiento con cobre disminuye la viabilidad de la Drosophila. La disminución de la sobrevida estuvo asociada a un aumento y una disminución de los registros de actividad motora en las etapas tempranas y tardías de la intoxicación respectivamente. Por último, se evaluó el papel del sistema de neurotransmisión dopaminérgico sobre las alteraciones conductuales inducidas por el cobre. El tratamiento con el precursor de la dopamina, L-dopa, indujo un aumento de la actividad motora similar al inducido por el cobre. Por el contrario, el tratamiento con Flufenazina, un antagonista de los receptores dopaminérgicos D2, fue capaz de impedir las alteraciones conductuales en todas las edades evaluadas. Estos resultados sugieren que la Drosophila melanogaster podría ser empleada como modelo para el estudio de posibles intervenciones con potencial terapéutico en la enfermedad de Wilson.

Wilson disease is a hereditary disorder caused by mutations of the ATP7B gene, which leads to intoxication with copper as a result of an unbalance of copper homeostasis. The clinical manifestations resulting from this intoxication are related to the affectation of liver and the encephalon in most cases. Several animal models are currently available for the study of the malady. However, in such models no neurological symptoms are observed, which limits their use for the study of pathogenic effects of this disease on the central nervous system. The aim of the present study was to evaluate if copper feeding could induce a disease state in Drosophila melanogaster to model Wilson disease. The effect of the feeding of copper at the doses of 31 µM and 47 µM on the survival was initially evaluated. Next, behavioral experiments were conducted to determine whether the motor performance was altered by the 47 µM concentration. The results suggest that copper treatment decreases the viability of the flies. In addition, the decrease of viability was associated to an increase and decrease of spontaneous motor activity at early and late stages of the intoxication, respectively. Finally, the role of the dopaminergic neurotransmission system on the observed motor alterations was evaluated. The dopamine precursor L-dopa increased motor activity. In contrast, D2 receptor antagonist, Fluphenazine, was able to block both the increase and decrease of motor activity scores induced by copper. These results suggest that Drosophila melanogaster could be used as a model organism for the study of possible interventions with potential neuroprotective effects in Wilson disease.

influence of different storage conditions on the stability of amitriptyline and fluphenazine in biological specimens

The aim of this research is to study the effect of different storage conditions [different temperatures and formalin preservation] on the stability of amitriptyline and fluphenazine in some biological samples. The LD[50] of amitriptyline and fluphenazine were administrated orally to rabbits which were sacrificed two hours after administration of the drugs. The tissues were stored at different conditions for six months. U.V. Spectrophotometer was used for estimation of the drugs at different periods. The results revealed that both amitriptyline and fluphenazine were rapidly declined in samples stored at room temperature [25 - 38°C]. It could not be detected in brain and liver samples at the end of three weeks, in the kidney at the end of four weeks and in plasma at the end of six weeks. While fluphenazine could not be detected in the brain at the end of three weeks, in the kidney and liver at the end of four weeks and in the plasma at the end of six weeks. At fridge temperature [5°C], amitriptyline could not be detected in brain and liver at the end of four weeks, in kidney samples at the end of six weeks. While fluphenazine could not be detected in brain at the end of four weeks, in kidney and liver at the end of six weeks, in plasma both of them couldn't be detected at the end of eight weeks. At freezer temperature [-20°C], amitriptyline could be detected up to the end of six months of the storage in the different samples with different relative recovery percent 80%, 75%, 69.57%, and 63.00% [for plasma, kidney, brain and liver samples respectively]. While fluphenazine could be detected up to the end four months of the storage in the plasma, kidney, and liver with different relative recovery percent [19.77%, 13.88%, and 11.56% respectively]. In brain fluphenazine could be detected up to the end of twelve weeks with a relative recovery percent of 15.76%. In samples preserved in 10% formalin solution, amitriptyline could be detected up to the end of six months of the storage in the different samples with high relative recovery percents [90.81%, 90.18%, 87.84% and 86.14%] for the kidney, plasma, brain, and liver respectively. While fluphenazine could not be detected in brain samples at the end of six weeks. It could not be detected in liver, kidney, and plasma at the end of eight weeks of the storage. In conclusion amitriptyline is stable in tissues stored at freezer [-20°C] and that preserved in formalin solution. While fluphenazine is stable in tissues stored at freezer [-20°C] for sometime, but it is not stable in the samples stored at room temperature, fridge temperature, and in samples preserved in formalin solution

Chronic effect of antidopaminergic drugs or estrogen on male Wistar rat lactotrophs and somatotrophs

The aim of the present study was to evaluate the effect of antidopaminergic agents on the somatotrophs in the presence of hyperprolactinemia. Adult male Wistar rats were divided into 6 groups: a control group and five groups chronically treated (60 days) with haloperidol, fluphenazine, sulpiride, metoclopramide or estrogen. Somatotrophs and lactotrophs were identified by immunohistochemistry and the data are reported as percent of total anterior pituitary cells counted. The drugs significantly increased the percentage of lactotrophs: control (mean + or - SD) 21.3 + or - 4.4, haloperidol 27.8 + or - 2.2, fluphenazine 34.5 + or - 3.6, sulpiride 32.7 + or - 3.5, metoclopramide 33.4 + or - 5.5 and estrogen 42.4 + or - 2.8. A significant reduction in somatotrophs was observed in animals treated with haloperidol (23.1 + or - 3.0), fluphenazine (22.1 + or - 1.1) and metoclopramide (24.2 + or - 3.0) compared to control (27.3 + or - 3.8), whereas no difference was observed in the groups treated with sulpiride (25.0 + or - 2.2) and estrogen (27.1 + or - 2.8). In the groups in which a reduction occurred, this may have simply been due to dilution, secondary to lactotroph hyperplasia. In view of the duplication of the percentage of prolactin-secreting cells, when estrogen was applied, the absence of a reduction in the percent of somatotrophs suggests a replication effect on this cell population. These data provide additional information about the direct or indirect effect of drugs which, in addition to interfering with the dopaminergic system, may act on other pituitary cells as well as on the lactotrophs.

Effect of estrogen and neuroleptics on prolactin secretion and immunoreactive prolactin cells

The use of estrogen and dopamine receptor antagonists is associated with elevated prolactin levels and, in rats, chronic estrogen treatment is also associated with lactotroph proliferation. In this study, haloperidol, fluphenazine, sulpiride and metoclopramide, alone or combined with estradiol, were administered to Wistar rats. Pituitary weight, serum prolactin levels and percent of immunoreactive prolactin cells in the anterior pituitary glands were determined at the end of 60 days of treatment. The pituitary weight of rats treated with estrogen alone or in combination with other drugs was significantly higher than the control group. The serum prolactin level was higher than the upper confidence limit in all but three of the 90 treated rats. While in the control group the percent of immunoreactive prolactin cells was 20 per cent, administration of the neuroleptic drugs and metoclopramide increased this percent to approximately 30 per cent, and estrogen alone or in combination with one of the neuroleptic drugs increased it to approximately 40 per cent. The results presented here demonstrate the elationship between prolactin secretion and prolactin cell number when different neuroleptics and related drugs are used.

Embarazo en usuarias de decanoato de flufenazina: descripción de 22 casos / Pregnancy and flufenazine decanoate

Generalmente en las pacientes usuarias de neurolépticos de depósito se mantiene un estricto control de la fertilidad, en caso de ocurrencia de embarazo se recomienda la suspensión del fármaco de depósito puesto que nose ha establecido su inocuidad durante la gestación. Se presentan 22 pacientes usuarias de decanoato de flufenazina que presentaron 25 gestaciones. Se trata de pacientes en control en la Clínica de Psicofármacos del Servicio de Psiquiatría del Hospital de Valdivia entre 1979 y 1989. Se analizan la historia psiquiátrica, indicaciones farmacológicas, antecedentes, evolución de la gestación y parto, condición del recién nacido, reagudizaciones y evolución posterior de las pacientes. De acuerdo a las experiencias analizadas se sugieren medidas terapéuticas en caso de ocurrencia de gestación en pacientes usuarias de neuroléptico de depósito

Participaçäo colinérgica nos efeitos das drogas antipsicóticas na febre devida a um pirogênio bacteriano / Cholinergic participation in effects of antipsichotics drugs in the fever elicited by one bacterial pyrogen

Os resultados mostraram que a associaçäo de atropina às drogas antipsicóticas produziu reduçäo da resposta febril, quando clorpromazina e flufenazina foram os antipsicóticos utilizados. A associaçäo de atropina ao haloperidol e ao pimozide potencializou a resposta febril

Possível envolvimento dopaminérgico na resposta febril induzida pelo lipopolissacarídeo de E. coli, no coelho / Dopaminergic involvement in the fever response induced in rabbits by lipopolysaccharide from E. coli

Drogas neurolépticas como a clorpromazina (CLOR), flufenazina (Flu), haloperidol (HAL) e pinozide (Pim), conhecidas por atuarem como antipsicóticas e bloqueadoras de receptores dopaminérgicos centrais, inibiram parcialmente a resposta febril induzida pelo lipopolissacarídeo de E. coli (LPS), um pirogênio bacteriano, em coelhos. Os presentes resultados sugerem que os receptores dopaminérgicos centrais podem estar envolvidos nessa resposta febril, muito embora essa mobilizaçäo näo represente o principal mecanismo