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1.
Toxicol Sci ; 170(1): 157-166, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30923810

RESUMO

3, 4-Methylenedioxymethamphetamine (MDMA) is a hallucinogenic amphetamine derivative. The acute effects of MDMA are hyperthermia, hyperactivity, and behavioral changes, followed by long-term serotonergic neurotoxicity in rats and primates. However, the underlying mechanisms of MDMA neurotoxicity remain elusive. We reported that pretreatment of rats with Ro 4-1284, a reversible inhibitor of the vesicular monoamine transporter 2 (VMAT2), reduced MDMA-induced hyperactivity in rats, abolished the hyperthermic response, and the long-term neurotoxicity. Current studies focused on the effects of co- and/or postinhibition of VMAT2 on the acute and chronic effects of MDMA and on the dose-response relationship between MDMA-induced elevations in body temperature and subsequent reductions in indolamine concentrations. Sprague Dawley rats were treated with MDMA (20, 25, or 27.5 mg/kg sc), and either co- and/or posttreatment with the VMAT2 inhibitor (10 mg/kg ip). Rats simultaneously treated with Ro 4-1284 and MDMA exhibited a more rapid increase in body temperature compared to just MDMA. However, the duration of the elevated body temperature was significantly shortened (approximately 3 h vs approximately 8 h, respectively). A similar body temperature response was observed in rats posttreated (7 h after MDMA) with Ro 4-1284. Despite decreases in the area under the curve (Δtemp X time) of body temperature caused by Ro 4-1284, there were no significant differences in the degree of indolamine depletion between any of the MDMA-treated groups. The results suggest that the neuroprotective effects of VMAT2 inhibition is likely due to the indirect monoamine depleting effects of the Ro 4-1284 pretreatment, rather than by the direct inhibition of VMAT2 function.


Assuntos
2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , N-Metil-3,4-Metilenodioxianfetamina/toxicidade , Síndromes Neurotóxicas/prevenção & controle , Proteínas Vesiculares de Transporte de Monoamina/antagonistas & inibidores , Animais , Temperatura Corporal/efeitos dos fármacos , Febre/induzido quimicamente , Febre/tratamento farmacológico , Indóis/metabolismo , Masculino , Síndromes Neurotóxicas/etiologia , Ratos , Ratos Sprague-Dawley
2.
ACS Chem Neurosci ; 6(11): 1802-12, 2015 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-26322962

RESUMO

Tonic dopamine (DA) levels influence the activity of dopaminergic neurons and the dynamics of fast dopaminergic transmission. Although carbon fiber microelectrodes and fast-scan cyclic voltammetry (FSCV) have been extensively used to quantify stimulus-induced release and uptake of DA in vivo and in vitro, this technique relies on background subtraction and thus cannot provide information about absolute extracellular concentrations. It is also generally not suitable for prolonged (>90 s) recordings due to drift of the background current. A recently reported, modified FSCV approach called fast-scan controlled-adsorption voltammetry (FSCAV) has been used to assess tonic DA levels in solution and in the anesthetized mouse brain. Here we describe a novel extension of FSCAV to investigate pharmacologically induced, slowly occurring changes in tonic (background) extracellular DA concentration, and phasic (stimulated) DA release in brain slices. FSCAV was used to measure adsorption dynamics and changes in DA concentration (for up to 1.5 h, sampling interval 30 s, detection threshold < 10 nM) evoked by drugs affecting DA release and uptake (amphetamine, l-DOPA, pargyline, cocaine, Ro4-1284) in submerged striatal slices obtained from rats. We also show that combined FSCAV-FSCV recordings can be used for concurrent study of stimulated release and changes in tonic DA concentration. Our results demonstrate that FSCAV can be effectively used in brain slices to measure prolonged changes in extracellular level of endogenous DA expressed as absolute values, complementing studies conducted in vivo with microdialysis.


Assuntos
Corpo Estriado/metabolismo , Dopamina/metabolismo , Técnicas Eletroquímicas/métodos , Espaço Extracelular/metabolismo , Técnicas de Cultura de Tecidos/métodos , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Anfetamina/farmacologia , Animais , Carbono , Fibra de Carbono , Cocaína/farmacologia , Corpo Estriado/efeitos dos fármacos , Dopaminérgicos/farmacologia , Estimulação Elétrica/instrumentação , Estimulação Elétrica/métodos , Técnicas Eletroquímicas/instrumentação , Espaço Extracelular/efeitos dos fármacos , Levodopa/farmacologia , Microeletrodos , Pargilina/farmacologia , Ratos Wistar , Técnicas de Cultura de Tecidos/instrumentação
3.
Toxicol Sci ; 143(1): 209-19, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25370842

RESUMO

3,4-(±)-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a ring-substituted amphetamine derivative with potent psychostimulant properties. The neuropharmacological effects of MDMA are biphasic in nature, initially causing synaptic monoamine release, primarily of serotonin (5-HT). Conversely, the long-term effects of MDMA manifest as prolonged depletions in 5-HT, and reductions in 5-HT reuptake transporter (SERT), indicative of serotonergic neurotoxicity. MDMA-induced 5-HT efflux relies upon disruption of vesicular monoamine storage, which increases cytosolic 5-HT concentrations available for release via a carrier-mediated mechanism. The vesicular monoamine transporter 2 (VMAT2) is responsible for packaging monoamine neurotransmitters into cytosolic vesicles. Thus, VMAT2 is a molecular target for a number of psychostimulant drugs, including methamphetamine and MDMA. We investigated the effects of depressed VMAT2 activity on the adverse responses to MDMA, via reversible inhibition of the VMAT2 protein with Ro4-1284. A single dose of MDMA (20 mg/kg, subcutaneous) induced significant hyperthermia in rats. Ro4-1284 (10 mg/kg, intraperitoneal) pretreatment prevented the thermogenic effects of MDMA, instead causing a transient decrease in body temperature. MDMA-treated rats exhibited marked increases in horizontal velocity and rearing behavior. In the presence of Ro4-1284, MDMA-mediated horizontal hyperlocomotion was delayed and attenuated, whereas rearing activity was abolished. Finally, Ro4-1284 prevented deficits in 5-HT content in rat cortex and striatum, and reduced depletions in striatal SERT staining, 7 days after MDMA administration. In summary, acute inhibition of VMAT2 by Ro4-1284 protected against MDMA-mediated hyperthermia, hyperactivity, and serotonergic neurotoxicity. The data suggest the involvement of VMAT2 in the thermoregulatory, behavioral, and neurotoxic effects of MDMA.


Assuntos
Encéfalo/efeitos dos fármacos , Alucinógenos , N-Metil-3,4-Metilenodioxianfetamina , Síndromes Neurotóxicas/prevenção & controle , Neurônios Serotoninérgicos/efeitos dos fármacos , Serotonina/metabolismo , Proteínas Vesiculares de Transporte de Monoamina/antagonistas & inibidores , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Regulação da Temperatura Corporal/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/fisiopatologia , Modelos Animais de Doenças , Febre/induzido quimicamente , Febre/fisiopatologia , Febre/prevenção & controle , Hipercinese/induzido quimicamente , Hipercinese/prevenção & controle , Hipercinese/psicologia , Masculino , Atividade Motora/efeitos dos fármacos , Síndromes Neurotóxicas/metabolismo , Síndromes Neurotóxicas/fisiopatologia , Síndromes Neurotóxicas/psicologia , Ratos Sprague-Dawley , Neurônios Serotoninérgicos/metabolismo , Neurônios Serotoninérgicos/patologia , Fatores de Tempo , Proteínas Vesiculares de Transporte de Monoamina/metabolismo
4.
Neuroscience ; 101(4): 1063-9, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11113355

RESUMO

The vesicular monoamine transporter in the brain can sequester the neurotoxin 1-methyl-4-phenylpyridinium into synaptic vesicles and protect catecholamine-containing neurons from degeneration. Mouse nigrostriatal dopaminergic neurons, and to a lesser extent locus coeruleus noradrenergic neurons, are vulnerable to toxicity produced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. The present study sought to determine whether pharmacological inactivation of the vesicular monoamine transporter in the brain would enhance the degeneration of substantia nigra dopaminergic neurons and locus coeruleus noradrenergic neurons in 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine-treated animals. Mice were treated subacutely with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine alone, or in combination with vesicular monoamine transporter inhibitors (tetrabenazine or Ro4-1284), and 10-24 days later striatal dopamine and cortical norepinephrine levels were measured using chromatographic methods. In the same animals, substantia nigra and locus coeruleus catecholaminergic neurons were counted using tyrosine hydroxylase immunohistochemical staining with computer imaging techniques. Mice in which pharmacological blockage of the vesicular monoamine transporter enhanced the effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in the depletion of striatal dopamine concentrations also exhibited enhanced degeneration of substantia nigra dopaminergic neurons. In the same animals, however, vesicular monoamine transporter blockade did not enhance the effects of 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine in the locus coeruleus noradrenergic system. These data are consistent with the hypothesis that the vesicular monoamine transporter can protect catecholamine-containing neurons from 1-methyl-4-phenylpyridinium-induced degeneration by sequestration of the toxin within brain vesicular monoamine transporter-containing synaptic vesicles. Since the amount of vesicular monoamine transporter in locus coeruleus neurons is more than in substantia nigra neurons, and because 1-methyl-4-phenylpyridinium is sequestered within locus coeruleus neurons to a far greater extent than within substantia nigra neurons, it may be that a greater amount of vesicular monoamine transporter inhibition is required for 1-methyl-4-phenylpyridinium to be toxic to locus coeruleus neurons than to substantia nigra dopaminergic neurons.


Assuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Glicoproteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana Transportadoras , Mesencéfalo/fisiopatologia , Degeneração Neural/induzido quimicamente , Degeneração Neural/fisiopatologia , Neurônios/fisiologia , Neuropeptídeos , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Contagem de Células , Sinergismo Farmacológico , Locus Cerúleo/fisiologia , Masculino , Mesencéfalo/patologia , Camundongos , Degeneração Neural/patologia , Neurônios/patologia , Norepinefrina/fisiologia , Tetrabenazina/farmacologia , Proteínas Vesiculares de Transporte de Aminas Biogênicas , Proteínas Vesiculares de Transporte de Monoamina
5.
Neuropsychobiology ; 41(3): 154-7, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-10754430

RESUMO

Dopamine depletion induced by administration of Ro4-1284 produces a condition of rapid and repeated eye blinking in rats. This condition mimics the human disorder, blepharospasm, which often accompanies parkinsonism and other dopamine deficiency disorders. When given a 3-week course of a compound (SR-3) developed from a specific ratio of two free polyunsaturated fatty acids - linoleic acid and alpha-linolenic acid - the eye blinking rate following administration of Ro4-1284 is reduced to saline and no drug control levels. These results suggest a favorable prospect for essential fatty acids in general, and SR-3 in particular, to provide an improved therapeutic option for the clinical management of benign essential blepharospasm.


Assuntos
Blefarospasmo/tratamento farmacológico , Blefarospasmo/prevenção & controle , Ácidos Graxos Essenciais/uso terapêutico , Ácidos Linolênicos/uso terapêutico , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol , Animais , Blefarospasmo/induzido quimicamente , Dopamina/deficiência , Dopamina/metabolismo , Combinação de Medicamentos , Ratos , Ratos Sprague-Dawley , Resultado do Tratamento , Gravação em Vídeo
6.
J Pharmacol Exp Ther ; 293(2): 336-42, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10773000

RESUMO

Dopamine neurons from various animal species differ in sensitivity to the neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or 1-methyl-4-phenylpyridinium (MPP(+)). Compared with striatal vesicles isolated from mice, those from rats have a higher density of the brain vesicular monoamine transporter (VMAT2) and a greater ability to sequester MPP(+), suggesting a larger storage capacity for MPP(+) in rat vesicles. In the present study, we examined whether striatal VMAT2-containing vesicles might provide protection against the neurotoxic effects of MPP(+) in vivo. Dose-response curves for striatally infused MPP(+) were determined in animals pretreated with or without a VMAT2 inhibitor. Ro 4-1284 administration (10 mg/kg i.p.; VMAT2 inhibitor) produced a 5-fold leftward shift in the MPP(+) dose-response curve and a significant lowering of the EC(50) concentration for MPP(+)-induced damage. These findings provide evidence for a substantial accumulation of MPP(+) in VMAT2-containing vesicles in vivo in the rat striatum and support the hypothesis that MPP(+) sequestration in vesicles can provide protection against its toxic actions. In mice, VMAT2 inhibition did not reliably enhance toxicity produced by a striatal infusion of MPP(+) or by systemic administration of MPTP. These data suggest that vesicular sequestration of MPP(+) may be of less importance in mice than in rats as relates to protection from the toxin. The present results also reveal that although VMAT2 inhibition enhanced striatal MPP(+) toxicity in the rat, the potency of MPP(+) in the rat striatum was less than that in mouse striatum. This implies that there are other factors that either exacerbate MPP(+) toxicity in the mouse or attenuate MPP(+) toxicity in rats.


Assuntos
1-Metil-4-fenilpiridínio/toxicidade , Química Encefálica/efeitos dos fármacos , Glicoproteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana Transportadoras , Neostriado/metabolismo , Neuropeptídeos , Neurotoxinas/toxicidade , Neurotransmissores/antagonistas & inibidores , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Relação Dose-Resposta a Droga , Masculino , Camundongos , Microdiálise , Neostriado/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Especificidade da Espécie , Vesículas Sinápticas/efeitos dos fármacos , Vesículas Sinápticas/metabolismo , Fatores de Tempo , Proteínas Vesiculares de Transporte de Aminas Biogênicas , Proteínas Vesiculares de Transporte de Monoamina
7.
J Neurochem ; 74(4): 1656-65, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10737624

RESUMO

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.


Assuntos
Corpo Estriado/citologia , Dopamina/fisiologia , Malonatos/farmacologia , Proteínas de Membrana Transportadoras , Neurônios/metabolismo , Neuropeptídeos , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Inibidores da Captação Adrenérgica/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/fisiologia , Radioisótopos de Carbono , Células Cultivadas , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Dopamina/farmacocinética , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Radicais Livres/metabolismo , Ácido Láctico/metabolismo , Masculino , Glicoproteínas de Membrana/metabolismo , Camundongos , Microdiálise , Degeneração Neural/induzido quimicamente , Degeneração Neural/metabolismo , Neurônios/efeitos dos fármacos , Doença de Parkinson/metabolismo , Terminações Pré-Sinápticas/efeitos dos fármacos , Vesículas Sinápticas/metabolismo , Tetrabenazina/farmacologia , Trítio , Proteínas Vesiculares de Transporte de Aminas Biogênicas , Ácido gama-Aminobutírico/metabolismo , Ácido gama-Aminobutírico/farmacocinética
8.
Eur J Pharmacol ; 370(1): 23-6, 1999 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-10323276

RESUMO

Tramadol is a centrally acting opioid analgesic whose mechanism of action could also involve an increase in central serotoninergic transmission. Thus, tramadol inhibits synaptosomal serotonin (5-hydroxytryptamine, 5-HT) reuptake and induces tritium release from [3H]5-HT-preloaded slices. We investigated the effect of (+/-)-tramadol in release studies with superfused rat brain cortex synaptosomes preloaded with [3H]5-HT. Tramadol had no releasing effect up to 30 microM, whereas at 10 microM tramadol significantly inhibited by 45% D-fenfluramine-induced [3H]5-HT release. At 100 microM, tramadol showed a slight releasing effect in the absence or in the presence of pargyline, which was not augmented in synaptosomes pre-exposed to Ro 04-1284 (2-ethyl-1,3,4,6,7,11b-hexahydro-3-isobutyl-9,10-dimethoxy-2H-benzo [a]quinolizin-2-ol hydrochloride), a reserpine-like compound that enhances cytoplasmic 5-HT levels. In summary, (+/-)-tramadol behaved as a classical 5-HT uptake blocker (like citalopram) and not as a substrate of the 5-HT carrier with indirect 5-HT mimetic properties (like D-fenfluramine).


Assuntos
Analgésicos Opioides/farmacologia , Córtex Cerebral/efeitos dos fármacos , Inibidores de Captação de Serotonina/farmacologia , Serotonina/metabolismo , Sinaptossomos/efeitos dos fármacos , Tramadol/farmacologia , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Células Cultivadas , Córtex Cerebral/metabolismo , Fenfluramina/farmacologia , Masculino , Ratos , Sinaptossomos/metabolismo
9.
Eur J Pharmacol ; 365(1): 27-34, 1999 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-9988119

RESUMO

Ro4-1284 (2-Ethyl-1,3,4,6,7,11b-hexahydro-3-isobutyl-9,10-dimethoxy-2H-benzo[a] quinolizin-2-ol hydrochloride), a benzoquinolizine, is a potent dopamine depletion agent whose acute and chronic administration results in a (1) deterioration of learning in the Morris Water Maze and passive avoidance tasks, (2) decrease in locomotion and rearing, (3) intense hypothermia, and (4) decrease in the percentage of polyunsaturated fatty acids and an increase in the level of cholesterol in neuronal membranes. Pretreatment with a specific mixture of free polyunsaturated fatty acids prevents most of the behavioral, physiological, and biochemical effects of Ro4-1284 except for rearing. We propose that the dopamine-mediated functions tested in this study are dependent on the interaction of intact dopamine D1 and D2 receptors. Rearing, which is controlled only by dopamine D1 receptors, remained, therefore, unaffected. Our hypothesis is that SR-3 exerts its beneficial effects by normalizing the structure and function of the neuronal membrane and by restoring dopamine D2 receptor functions.


Assuntos
2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Ácidos Graxos Insaturados/farmacologia , Deficiências da Aprendizagem/prevenção & controle , Análise de Variância , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Temperatura Corporal/efeitos dos fármacos , Colesterol/metabolismo , Ácidos Graxos Insaturados/metabolismo , Deficiências da Aprendizagem/induzido quimicamente , Aprendizagem em Labirinto/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
10.
Brain Res ; 815(1): 21-8, 1999 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-9974118

RESUMO

Ponto-geniculo-occipital (PGO) waves are spontaneously occurring field potentials recorded in the dorsal lateral geniculate nucleus (LGN) just prior to and during rapid eye movement (REM) sleep. Facilitated discharge rates of LGN neurons are associated with PGO waves. In kittens during the critical period of visual system development, both visual experience and PGO waves appear capable of influencing the course of development through activity-dependent mechanisms. Retinal innervation of LGN segregates into eye-specific laminae and is critical to supporting the role of binocular visual experience in development. We sought to determine whether neuronal activity associated with PGO waves also exhibits lamina specificity. PGO wave-related discharges were examined in LGN neurons identified as to lamina location in adult cats administered urethane anesthesia and the reserpine-like compound, RO4-1284. Spontaneous activity of LGN neurons was related to the occurrence of PGO-like waves in all cells studied. No factors could be found that differentiated lamina location and PGO wave-related discharges. We conclude that the PGO wave influence on neuronal activity in the visual system is fundamentally different from that derived from visual experience. The implications of this difference for the role of the two sources of activation in the control of neural activity in development are discussed.


Assuntos
Corpos Geniculados/citologia , Neurônios/fisiologia , Lobo Occipital/citologia , Ponte/citologia , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Fatores Etários , Animais , Gatos , Feminino , Corpos Geniculados/fisiologia , Vias Neurais , Lobo Occipital/fisiologia , Ponte/fisiologia , Sono REM/fisiologia , Córtex Visual/citologia , Córtex Visual/fisiologia
11.
J Neurosci ; 18(6): 1979-86, 1998 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-9482784

RESUMO

Amphetamine (AMPH) inhibits uptake and causes release of dopamine (DA) from presynaptic terminals. AMPH can act on both vesicular storage of DA and directly on the dopamine transporter (DAT). To assess the relative importance of these two processes, we have examined the releasing actions of AMPH in mice with a genetic deletion of the DAT. The sequence of actions of AMPH has been determined by following the real time changes of DA in the extracellular fluid of intact tissue with fast scan cyclic voltammetry. In striatal slices from wild-type mice, AMPH causes a gradual (approximately 30 min) increase in extracellular DA, with a concomitant disappearance of the pool of DA available for depolarization-evoked release. Conversely, in slices from mice lacking the DAT, although a similar disappearance of electrically stimulated DA release occurs, extracellular DA does not increase. Similarly, microdialysis measurements of DA after AMPH in freely moving animals show no change in mice lacking the DAT, whereas it increases 10-fold in wild-type mice. In contrast, redistribution of DA from vesicles to the cytoplasm by the use of a reserpine-like compound, Ro4-1284, does not increase extracellular DA in slices from wild-type animals; however, subsequent addition of AMPH induces rapid (<5 min) release of DA. Thus, the DAT is required for the releasing action, but not the vesicle-depleting action, of AMPH on DA neurons, and the latter represents the rate-limiting step in the effects of AMPH. Furthermore, these findings suggest that in the absence of pharmacological manipulation, such as the use of amphetamine, endogenous cytoplasmic DA normally does not reach sufficient concentrations to reverse the DAT.


Assuntos
Anfetamina/farmacologia , Proteínas de Transporte/genética , Estimulantes do Sistema Nervoso Central/farmacologia , Glicoproteínas de Membrana , Proteínas de Membrana Transportadoras , Camundongos Knockout/genética , Proteínas do Tecido Nervoso , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Dopamina/metabolismo , Antagonistas de Dopamina/farmacologia , Proteínas da Membrana Plasmática de Transporte de Dopamina , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microdiálise , Sulpirida/farmacologia , Tetrabenazina/farmacologia
12.
J Nucl Med ; 37(12): 2011-6, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8970525

RESUMO

UNLABELLED: Para-[18F]fluorobenzylguanidine ([18F]PFBG) is a newly developed tracer for imaging myocardial sympathetic neuronal innervation. This study investigated the uptake and retention mechanisms of [18F]PFBG in perfused, isolated rat heart. METHODS: Fluorine-18-PFBG was administered to working rat hearts within the perfusion medium at a constant activity concentration (1.5-2 MBq/liter) for 8 min, followed by a washout period (50 min). External scintillation probes with coincidence detection circuitry were used to measure myocardial radioactivity. Six groups of hearts (n = 6, except in Group 6) were studied: (Group 1) control; (Group 2) 100 nM desipramine (DMI); (Group 3) 0.8 microM SKF550; (Group 4) DMI + SKF550; (Group 5) SKF550 + 1.0 microM Ro 4-1284; and (Group 6) SKF550 with DMI chase at 30 min (n = 4). RESULTS: Groups 2, 3 and 4 showed a mean reduction of 19% (uptake-1 blockade), 58% (uptake-2 blockade) and 95% (uptake-1 and uptake-2 blockade) in uptake rates, respectively, compared with control (p < 0.01). A further 33% reduction in the uptake rate was noted with vesicular transport inhibition (Group 5 compared with 3, p = 0.054). Biphasic clearance consisting of rapid (T1/2 = 5.32 +/- 1.1 min) and slow (T1/2 = 35.2 +/- 9.6 min) components were noted in control hearts. The rapid (T1/2 = 1.6 +/- 0.3 min) and slow (T1/2 = 10.9 +/- 1.4 min) clearance rates were accelerated (p < 0.0001) in Group 5 compared to control. DMI chase conditions (Group 6) caused an inhibition of [18F]PFBG washout (p = 0.004) suggesting a role for reverse transport through the uptake-1 carrier. CONCLUSION: Fluorine-18-PFBG is specifically accumulated by sympathetic nerve terminals. However, further work is recommended in humans to evaluate the potential implications of specific extraneuronal uptake of [18F]PFBG through the uptake-2 mechanism.


Assuntos
Fluorbenzenos , Guanidinas , Coração/diagnóstico por imagem , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Desipramina/farmacologia , Feminino , Fluorenos/farmacologia , Fluorbenzenos/farmacocinética , Guanidinas/farmacocinética , Coração/efeitos dos fármacos , Coração/inervação , Hemodinâmica/efeitos dos fármacos , Técnicas In Vitro , Cintilografia , Ratos , Ratos Sprague-Dawley , Sistema Nervoso Simpático/diagnóstico por imagem , Sistema Nervoso Simpático/metabolismo
13.
Neuropharmacology ; 35(11): 1627-34, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-9025111

RESUMO

Fast cyclic voltammetry (FCV) was used to measure electrically stimulated monoamine efflux in the rat ventral lateral geniculate nucleus (vLGN). The electrochemical characteristics of the released species resembled 5-HT but not dopamine or noradrenaline. Amine efflux was abolished by the sodium channel blocker tetrodotoxin (0.1 microM), Ro 4-1284 (1.0 microM), the fast-acting reserpine analogue, and removal of Ca2+ from the superfusate. Amine efflux was unaffected by the monoamine oxidase inhibitor clorgyline (0.1 microM). Of paroxetine (0.1 microM), desipramine (50 nM) and vanoxerine (0.5 microM), selective blockers of 5-HT, noradrenaline and dopamine uptake respectively, only paroxetine increased monoamine efflux (to 194 +/- 25%, mean +/- SEM) and prolonged the removal half-life (to 638 +/- 105%). The non-specific 5-HT1 antagonist methiothepin (0.2 microM) increased 5-HT efflux on long (20 pulses at 20 Hz) but not short trains (20 pulses at 100 Hz). When tested on pseudo-one-pulse stimulations (5 pulses, 100 Hz), the selective 5-HT1A agonist 8-OHDPAT (1.0 microM) had no effect. CP 93129 (0.3 microM), the selective 5-HT1B agonist, decreased 5-HT efflux to 37 +/- 4% of control and was antagonised by the 5-HT1B blocker isamoltane (0.5 microM) and by the 5-HT1D/B antagonist GR 127935 (50 nM). The preferential 5-HT1D agonist sumatriptan (0.5 microM) also decreased 5-HT efflux, to 55 +/- 6% and was antagonised by GR 127935 (50 nM) but not isamoltane (0.5 microM). These results suggest that 5-HT released in the vLGN can be measured by FCV. Furthermore, released 5-HT is taken up by the 5-HT transporter and may be under the influence of 5-HT1B and 5-HT1D autoreceptors.


Assuntos
Autorreceptores/metabolismo , Corpos Geniculados/metabolismo , Receptores de Serotonina/metabolismo , Serotonina/metabolismo , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Clorgilina/farmacologia , Estimulação Elétrica , Meia-Vida , Masculino , Microeletrodos , Inibidores da Monoaminoxidase/farmacologia , Ratos , Ratos Wistar , Inibidores de Captação de Serotonina/farmacologia , Tetrodotoxina/farmacologia
14.
Microvasc Res ; 50(2): 215-22, 1995 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8538501

RESUMO

A method providing a biochemical index for the evaluation of promoters or inhibitors of angiogenesis in the chick chorioallantoic membrane (CAM) is here described and validated. This method is based on the determination of collagenous protein synthesis which takes place during new vessel formation. Validation was done by comparing collagenous protein synthesis to morphological methods of determining vascular density either by counting the number of vessels intersecting three concentric rings or by computer-assisted image analysis. Five compounds which promote or inhibit angiogenesis in the CAM were used for this purpose. The protein kinase C activator 4-beta-phorbol 12-myristate 13-acetate and alpha-thrombin increased collagenous protein synthesis and these results correlated with those obtained by using the two morphological methods mentioned above. Similarly, the inhibitors of angiogenesis, Ro318220, tricyclodecan-9-yl xanthate (D609), and 8,9-dihydroxy-7-methyl-benzo[b]quinolizinium bromide (GPA1734), reduced collagenous protein synthesis and vascular density (determined by image analysis or by counting the number of vessels intersecting three concentric rings) to a comparable degree. These results indicate that collagenous protein synthesis can be used as a reliable, reproducible, and unbiased index of angiogenesis in the chick chorioallantoic membrane.


Assuntos
Colágeno/biossíntese , Neovascularização Fisiológica/efeitos dos fármacos , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/análogos & derivados , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Antineoplásicos/farmacologia , Biomarcadores/análise , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Embrião de Galinha , Córion/irrigação sanguínea , Norbornanos , Ésteres de Forbol/farmacologia , Tiocarbamatos , Tionas/farmacologia , Trombina/farmacologia
15.
Lab Invest ; 71(4): 575-82, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7526036

RESUMO

BACKGROUND: The formation of a basement membrane is the last step in the development of a new blood vessel. Matrigel, a laminin-rich reconstituted basement membrane matrix induces the differentiation of endothelial cells into capillary-like structures. EXPERIMENTAL DESIGN: The effect of inhibitors of basement membrane collagen synthesis, tricyclodecan-9-yl xanthate (D609) and 8,9-dihydroxy-7-methyl-benzo[b] quinolizinium bromide (GPA 1734), was investigated on endothelial cell tube formation on Matrigel in vitro and in an angiogenesis assay in C57 black mice in vivo. RESULTS: D609 and GPA 1734 caused a dose-dependent decrease in tube formation in vitro with complete inhibition at 50 micrograms/ml for D609 and 15 micrograms/ml for GPA 1734. The inhibitory effect on capillary tube formation by both agents was reversible. Tube formation correlated well with collagenous protein biosynthesis. Parallel studies on endothelial cells cultured on plastic indicate that cell viability, proliferation, attachment, and morphology were not affected by the presence of these collagen inhibitors at doses that blocked tube formation and collagen biosynthesis. D609 and GPA 1734 also inhibited endothelial cell infiltration in response to SIKVAV in an in vivo angiogenesis model system. CONCLUSIONS: These results indicate that newly synthesised collagen is a prerequisite for expression of the endothelial cell phenotype for tube formation and that prevention of collagenous protein biosynthesis inhibits tube formation and angiogenesis in vivo.


Assuntos
2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/análogos & derivados , Antineoplásicos/farmacologia , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Colágeno/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Laminina , Neovascularização Patológica , Proteoglicanas , Tionas/farmacologia , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Membrana Basal/metabolismo , Membrana Basal/ultraestrutura , Adesão Celular/efeitos dos fármacos , Adesão Celular/fisiologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Feminino , Humanos , Camundongos , Norbornanos , Fenótipo , Tiocarbamatos
16.
Gen Pharmacol ; 25(5): 1039-43, 1994 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7835622

RESUMO

1. The reserpine-like agent, Ro4-1284 (2-hydroxy-2ethyl-3-isobutyl-9,10-dimethoxy-1,2,3,4,6,7-hexahydro- 11b-[H] benzo (a)quinolizine) releases [3H]noradrenaline ([3H]NA) from prelabelled superior cervical ganglion (cell bodies) and nictitating membrane (nerve endings) of the cat. 2. The potency of Ro 4-1284 29.0 microM was higher in the cell bodies than in the nerve endings. 3. In both tissues, exposure to the reserpine-like agent Ro 4-1284 induced a selective increase in the spontaneous outflow of [3H]DOPEG, while the [3H]OMDA metabolites to the release induced by Ro 4-1284 was very small. 4. The desamination is the preferential way of the metabolic inactivation of the [3H]NA released by the reserpine-like agent in both parts of the noradrenergic neuron.


Assuntos
2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Membrana Nictitante/efeitos dos fármacos , Norepinefrina/metabolismo , Gânglio Cervical Superior/efeitos dos fármacos , Animais , Gatos , Feminino , Técnicas In Vitro , Masculino , Membrana Nictitante/metabolismo , Gânglio Cervical Superior/metabolismo
17.
Naunyn Schmiedebergs Arch Pharmacol ; 348(5): 466-71, 1993 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8114945

RESUMO

The aim of the present work was to study the influence of tissue morphological characteristics on the neuronal release (and by inference the distribution) of tritiated and endogenous noradrenaline. Rat vas deferens and dog spleen capsule were loaded with 0.2 mumol/l 3H-noradrenaline, after inhibition of the noradrenaline metabolizing enzymes. Some preparations were washed out under control conditions (spontaneous efflux) and others were washed out in the presence of the releasing agents: 40 mumol/l of Ro 4-1284 (a reserpine-like compound), 100 mmol/l potassium or 100 mumol/l tyramine. The fractional rate of loss (efflux/tissue content) of each amine was determined and the ratio "endogenous amine/3H-noradrenaline" in the efflux and in the tissue were also calculated. The results showed no preferential release of one of the amines in the spleen capsule, whereas a preferential release of tritiated noradrenaline was observed in the vas deferens. The smooth muscle layer in the vas deferens was much thicker and more compact than that of the spleen capsule. The 3H-sorbitol space was smaller in the former than in the latter. We conclude that the morphological characteristics of the tissues contribute to the differences in 3H-noradrenaline distribution in the adrenergic varicosities of these preparations.


Assuntos
Norepinefrina/metabolismo , Baço/metabolismo , Ducto Deferente/metabolismo , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Cães , Técnicas In Vitro , Masculino , Microscopia Eletrônica , Músculo Liso/anatomia & histologia , Músculo Liso/metabolismo , Músculo Liso/ultraestrutura , Norepinefrina/farmacocinética , Ratos , Ratos Wistar , Sorbitol , Baço/anatomia & histologia , Ducto Deferente/anatomia & histologia , Ducto Deferente/ultraestrutura
18.
J Neurochem ; 61(2): 481-9, 1993 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8101557

RESUMO

A single dose of x-irradiation was applied on the cephalic end of newborn rats, and the alterations in the noradrenergic afferents to the cerebellum were studied 180 days later. A net increase in the noradrenaline content of cerebellum was found (122% of nonirradiated controls). The response of noradrenaline content to reserpine injection (0.9 mg/kg, i.p.) was similar in exposed and control rats. Likewise, the 3H release induced by Ro 4-1284 from cerebellar cortex slices labeled with [3H]noradrenaline was unmodified by x-rays, although a mild increase in the spontaneous efflux of 3H was found. The retention of 3H by the slices was reduced in exposed animals (58% of controls). Both the in vitro activity of tyrosine hydroxylase and the accumulation of L-3,4-dihydroxyphenylalanine (L-DOPA) were not significantly different between x-treated rats and controls. In contrast, monoamine oxidase activity was markedly reduced in x-irradiated cerebellum (38% of controls). The x-ray-induced decrease in cerebellar weight (-60%) resulted in marked increases in noradrenaline concentration (223%), tyrosine hydroxylase activity per milligram of protein (206%), and 3H retention (50%). The accumulation of L-DOPA per gram of tissue was also increased at every time considered. These data indicate that x-irradiation at birth produces a cerebellar loss not completely shared by the noradrenergic afferents, and a permanent imbalance between the noradrenergic afferent input and its target cells might eventually result.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Vias Aferentes/metabolismo , Animais Recém-Nascidos , Cerebelo/efeitos da radiação , Norepinefrina/fisiologia , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Envelhecimento , Animais , Córtex Cerebelar/efeitos dos fármacos , Córtex Cerebelar/metabolismo , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Feminino , Técnicas In Vitro , Cinética , Levodopa/metabolismo , Masculino , Ratos , Ratos Wistar , Reserpina/farmacologia , Trítio , Tirosina 3-Mono-Oxigenase/metabolismo
19.
J Cell Biochem ; 52(1): 92-106, 1993 May.
Artigo em Inglês | MEDLINE | ID: mdl-8320279

RESUMO

Type IV collagen-degrading activity was expressed in homogenates of Lytechinus pictus embryos during embryogenesis. Activity was concentrated 1,600-fold by ammonium sulfate fractionation, ion exchange, and gel chromatography and could not be activated further upon trypsin or organomercurial treatment. This enzyme activity could also degrade gelatin but had no affinity for type I, III, and V collagens. Activity was inhibited by addition of excess type IV collagen or gelatin, but was unaffected by addition of excess amounts of non-collagenous proteins of the extracellular matrix. Chelators such as 1,10-phenanthroline or Na2EDTA reduced activity to control levels. Inhibitors of plasmin and of serine and thiol proteases were without effect. Type IV collagen-degrading activity first became apparent at the stage of early mesenchyme blastula. It then increased by a small increment and remained stable up to the stage of late mesenchyme blastula, coinciding with first detection of collagen synthesis and the appearance of the archenteron. Thereafter, a sharp increase in activity was observed, concurrently with remodelling of the archenteron. Maximum activity was attained at prism stage and was retained throughout to pluteus-larva stage. The specific inhibitors of collagen biosynthesis 8,9-dihydroxy-7-methyl-benzo[b]quinolizinium bromide and tricyclodecane-9-yl xanthate arrested sea urchin embryo development at early blastula, prevented the invagination of the archenteron, and reverted the expression of type IV collagen-degrading activity to non-detectable levels. Removal of the inhibitors allowed embryos to gastrulate and express type IV collagen-degrading activity.


Assuntos
Colágeno/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/análogos & derivados , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Cromatografia em Gel , Colágeno/biossíntese , Gástrula/efeitos dos fármacos , Gástrula/metabolismo , Norbornanos , Ouriços-do-Mar/embriologia , Tiocarbamatos , Tionas/farmacologia
20.
J Pharmacol Exp Ther ; 265(1): 103-11, 1993 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8473998

RESUMO

The effects of reversible inhibitors of monoamine oxidase-A (moclobemide, Ro 41-1049, both 20 mg/kg, i.p., and brofaromine, 10 mg/kg, i.p.) on the outflow of dopamine (DA) and its metabolites (3,4-dihydroxyphenylacetic acid and homovanillic acid) as well as of 5-hydroxyindoleacetic acid was investigated by trans-striatal microdialysis in rats. These drugs markedly increased the level of DA in the dialysis fluid by 100% of basal values and concomitantly reduced the output of 3,4-dihydroxy-phenylacetic acid and homovanillic acid by 90%. The presence of tetrodotoxin in the perfusion fluid decreased the basal DA outflow and virtually abolished the rise in DA efflux after moclobemide administration. On the other hand, tetrodotoxin did not counteract the DA outflow induced by Ro 4-1284 (1 mg/kg, i.p.), a tetrabenazine derivative which rapidly releases DA from vesicles and causes a massive increase in the concentration of extravesicular amine. The injection of Ro 4-1284 30 min after moclobemide, brofaromine or Ro 41-1049 induced a 6-fold increase in DA outflow, which was accompanied by a transient increase in 3,4-dihydroxyphenylacetic acid levels. This latter effect was more marked for moclobemide than for the other two reversible inhibitors tested and was not observed in rats given the irreversible inhibitor clorgyline (5 mg/kg, i.p.). These results support the view that a large increase in the concentration of endogenous substrates in the cytosol might displace reversible monoamine oxidase-A inhibitors from the enzyme active sites. Therefore, the microdialysis technique seems to be a reliable in vivo method for assessing the degree of reversibility of monoamine oxidase inhibitors.


Assuntos
Benzamidas/farmacologia , Núcleo Caudado/efeitos dos fármacos , Citosol/metabolismo , Dopamina/metabolismo , Inibidores da Monoaminoxidase/farmacologia , Piperidinas/farmacologia , 2-etil-1,3,4,6,7,11b-hexaidro-3-isobutil-9,10-dimetoxi-2H-benzo(a)quinolizin-2-ol/farmacologia , Animais , Monoaminas Biogênicas/metabolismo , Núcleo Caudado/metabolismo , Masculino , Moclobemida , Ratos , Serotonina/metabolismo , Tetrodotoxina/farmacologia
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