RESUMO
NMDA type glutamate receptors (NMDARs) are best known for their role in synaptogenesis and synaptic plasticity. Much less is known about their developmental role before neurons form synapses. We report here that VEGF, which promotes migration of granule cells (GCs) during postnatal cerebellar development, enhances NMDAR-mediated currents and Ca(2+) influx in immature GCs before synapse formation. The VEGF receptor Flk1 forms a complex with the NMDAR subunits NR1 and NR2B. In response to VEGF, the number of Flk1/NR2B coclusters on the cell surface increases. Stimulation of Flk1 by VEGF activates Src-family kinases, which increases tyrosine phosphorylation of NR2B. Inhibition of Src-family kinases abolishes the VEGF-dependent NR2B phosphorylation and amplification of NMDAR-mediated currents and Ca(2+) influx in GCs. These findings identify VEGF as a modulator of NMDARs before synapse formation and highlight a link between an activity-independent neurovascular guidance cue (VEGF) and an activity-regulated neurotransmitter receptor (NMDAR).
Assuntos
Cerebelo/citologia , Neurônios/ultraestrutura , Receptores de N-Metil-D-Aspartato/fisiologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Quinases da Família src/metabolismo , Indutores da Angiogênese , Animais , Cálcio/metabolismo , Camundongos , Complexos Multiproteicos , Fosforilação , Receptores de Neurotransmissores , Sinapses , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Oxytocin is a neuromodulator with antidepressant-like effects. In vitro, oxytocin is rapidly cleaved by insulin-regulated aminopeptidase (IRAP). Oxytocin metabolites are known to exert strong central activities that are different from the effects of the parent molecule. Our goal is to investigate in vivo whether IRAP deletion modifies the antidepressant-like effects of oxytocin. Male and female C57Bl/6 mice, IRAP wild-type (IRAP(+/+)) and knock-out (IRAP(-/-)) mice were injected subcutaneously with saline, oxytocin or oxytocin combined with angiotensin IV. One hour after injection, immobility was timed during a 5 min forced swim that was preceded by an open field to study locomotor behaviour. Oxytocin induced antidepressant-like effects in male (0.25 mg/kg oxytocin) and female (0.15 mg/kg oxytocin) C57Bl/6 mice subjected to the forced swim test. Oxytocin did not influence locomotor behaviour in mice, as shown with the open field. These findings were reproduced in transgenic male (aged 3-6 months) and female (aged 12-18 months) IRAP(+/+) mice. However, the major findings of our study were that the antidepressant-like effect was reversed in angiotensin IV treated IRAP(+/+) mice and was completely absent in age- and gender-matched IRAP(-/-) mice. The lack of an antidepressant-like effect of oxytocin in young male and middle-aged female IRAP(-/-) mice attributes an important role to IRAP in mediating this effect.
Assuntos
Aminopeptidases/metabolismo , Antidepressivos/uso terapêutico , Depressão/tratamento farmacológico , Ocitocina/uso terapêutico , Fatores Etários , Angiotensina II/análogos & derivados , Angiotensina II/farmacologia , Animais , Antidepressivos/farmacologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/enzimologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Cistinil Aminopeptidase/deficiência , Cistinil Aminopeptidase/genética , Depressão/sangue , Depressão/patologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Comportamento Exploratório/efeitos dos fármacos , Feminino , Imipramina/uso terapêutico , Resposta de Imobilidade Tônica/efeitos dos fármacos , Insulina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Ocitocina/sangue , Ocitocina/farmacologiaRESUMO
L-Theanine, an ethylamide derivate of glutamate found in abundance in green tea, has been shown to exert beneficial actions in animal models for several neurological disorders. We here investigated for the first time the effect of L-theanine intake on seizure susceptibility using acute pilocarpine and pentylenetetrazol (PTZ) mouse models for studying, respectively, limbic seizures or primarily generalized seizures. Moreover, we studied the effect of l-theanine intake on extracellular hippocampal and cortical glutamate and gamma-aminobutyric acid (GABA) levels, using in vivo microdialysis. Feeding mice with a 4% L-theanine solution significantly decreased their susceptibility to pilocarpine-induced seizures whereas susceptibility to PTZ-induced seizures was increased. The latter effect was linked to decreased extracellular GABA concentrations in frontal cortex.
Assuntos
Glutamatos/farmacologia , Convulsões/tratamento farmacológico , Ácido gama-Aminobutírico/metabolismo , Animais , Modelos Animais de Doenças , GABAérgicos/metabolismo , Glutamatos/administração & dosagem , Ácido Glutâmico/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Microdiálise , Pentilenotetrazol/efeitos adversos , Pilocarpina/efeitos adversos , Convulsões/induzido quimicamente , Chá/químicaRESUMO
System x(c)- exchanges intracellular glutamate for extracellular cystine, giving it a potential role in intracellular glutathione synthesis and nonvesicular glutamate release. We report that mice lacking the specific xCT subunit of system x(c)- (xCT(-/-)) do not have a lower hippocampal glutathione content, increased oxidative stress or brain atrophy, nor exacerbated spatial reference memory deficits with aging. Together these results indicate that loss of system x(c)- does not induce oxidative stress in vivo. Young xCT(-/-) mice did however display a spatial working memory deficit. Interestingly, we observed significantly lower extracellular hippocampal glutamate concentrations in xCT(-/-) mice compared to wild-type littermates. Moreover, intrahippocampal perfusion with system x(c)- inhibitors lowered extracellular glutamate, whereas the system x(c)- activator N-acetylcysteine elevated extracellular glutamate in the rat hippocampus. This indicates that system x(c)- may be an interesting target for pathologies associated with excessive extracellular glutamate release in the hippocampus. Correspondingly, xCT deletion in mice elevated the threshold for limbic seizures and abolished the proconvulsive effects of N-acetylcysteine. These novel findings sustain that system x(c)-) is an important source of extracellular glutamate in the hippocampus. System x(c)(-) is required for optimal spatial working memory, but its inactivation is clearly beneficial to decrease susceptibility for limbic epileptic seizures.
Assuntos
Sistema y+ de Transporte de Aminoácidos/fisiologia , Espaço Extracelular/metabolismo , Ácido Glutâmico/metabolismo , Hipocampo/fisiologia , Sistema Límbico/fisiologia , Memória de Curto Prazo/fisiologia , Estresse Oxidativo/fisiologia , Convulsões/fisiopatologia , Percepção Espacial/fisiologia , Animais , Western Blotting , Encéfalo/anatomia & histologia , DNA/genética , Eletroencefalografia , Genótipo , Glutationa/metabolismo , Imuno-Histoquímica , Masculino , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microdiálise , Atividade Motora/fisiologia , Neuroglia/fisiologia , Desempenho Psicomotor/fisiologia , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Convulsões/induzido quimicamente , Convulsões/genéticaRESUMO
BACKGROUND: For successful translation to clinical stroke studies, the Stroke Therapy Academic Industry Round Table criteria have been proposed. Two important criteria are testing of therapeutic interventions in conscious animals and the presence of a co-morbidity factor. We chose to work with hypertensive rats since hypertension is an important modifiable risk factor for stroke and influences the clinical outcome. We aimed to compare the susceptibility to ischemia in hypertensive rats with those in normotensive controls in a rat model for induction of ischemic stroke in conscious animals. METHODS: The vasoconstrictor endothelin-1 was stereotactically applied in the vicinity of the middle cerebral artery of control Wistar Kyoto rats (WKYRs) and spontaneously hypertensive rats (SHRs) to induce a transient decrease in striatal blood flow, which was measured by the laser Doppler technique. Infarct size was assessed histologically by cresyl violet staining. Sensory-motor functions were measured at several time points using the neurological deficit score. Activation of microglia and astrocytes in the striatum and cortex was investigated by immunohistochemistry using antibodies against CD68/Iba-1 and glial fibrillary acidic protein. RESULTS AND CONCLUSIONS: The SHRs showed significantly larger infarct volumes and more pronounced sensory-motor deficits, compared to the WKYRs at 24 h after the insult. However, both differences disappeared between 24 and 72 h. In SHRs, microglia were less susceptible to activation by lipopolysaccharide and there was a reduced microglial activation after induction of ischemic stroke. These quantitative and qualitative differences may be relevant for studying the efficacy of new treatments for stroke in accordance to the Stroke Therapy Academic Industry Round Table criteria.
Assuntos
Isquemia Encefálica/metabolismo , Regulação para Baixo/fisiologia , Hipertensão/metabolismo , Lipopolissacarídeos/toxicidade , Microglia/metabolismo , Microglia/patologia , Acidente Vascular Cerebral/metabolismo , Animais , Isquemia Encefálica/genética , Isquemia Encefálica/patologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Corpo Estriado/patologia , Regulação para Baixo/efeitos dos fármacos , Endotelina-1/toxicidade , Predisposição Genética para Doença/etiologia , Hipertensão/complicações , Hipertensão/patologia , Lipopolissacarídeos/administração & dosagem , Masculino , Microglia/efeitos dos fármacos , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Acidente Vascular Cerebral/genética , Acidente Vascular Cerebral/patologiaRESUMO
BACKGROUND: Stroke remains one of the most common diseases with a serious impact on quality of life but few effective treatments exist. Mild hypothermia (33°C) is a promising neuroprotective therapy in stroke management. This study investigated whether a delayed short mild hypothermic treatment is still beneficial as neuroprotective strategy in the endothelin-1 (Et-1) rat model for a transient focal cerebral ischemia. Two hours of mild hypothermia (33°C) was induced 20, 60 or 120 minutes after Et-1 infusion. During the experiment the cerebral blood flow (CBF) was measured via Laser Doppler Flowmetry in the striatum, which represents the core of the infarct. Functional outcome and infarct volume were assessed 24 hours after the insult. In this sub-acute phase following stroke induction, the effects of the hypothermic treatment on apoptosis, phagocytosis and astrogliosis were assessed as well. Apoptosis was determined using caspase-3 immunohistochemistry, phagocytic cells were visualized by CD-68 expression and astrogliosis was studied by glial fibrillary acidic protein (GFAP) staining. RESULTS: Cooling could be postponed up to 1 hour after the onset of the insult without losing its positive effects on neurological deficit and infarct volume. These results correlated with the caspase-3 staining. In contrast, the increased CD-68 expression post-stroke was reduced in the core of the insult with all treatment protocols. Hypothermia also reduced the increased levels of GFAP staining, even when it was delayed up to 2 hours after the insult. The study confirmed that the induction of the hypothermia treatment in the Et-1 model does not affect the CBF. CONCLUSIONS: These data indicate that in the Et-1 rat model, a short mild hypothermic treatment delayed for 1 hour is still neuroprotective and correlates with apoptosis. At the same time, hypothermia also establishes a lasting inhibitory effect on the activation of astrogliosis.
Assuntos
Apoptose/efeitos dos fármacos , Isquemia Encefálica , Endotelina-1/efeitos adversos , Gliose/etiologia , Hipotermia Induzida/métodos , Estatística como Assunto , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Apoptose/fisiologia , Infarto Encefálico/etiologia , Isquemia Encefálica/induzido quimicamente , Isquemia Encefálica/complicações , Isquemia Encefálica/terapia , Caspase 3/metabolismo , Contagem de Células , Circulação Cerebrovascular/efeitos dos fármacos , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/terapia , Fluxometria por Laser-Doppler , Masculino , Transtornos dos Movimentos/etiologia , Exame Neurológico , Distribuição Aleatória , Ratos , Ratos Wistar , Fatores de TempoRESUMO
Malfunctioning of system x(c)(-), responsible for exchanging intracellular glutamate for extracellular cystine, can cause oxidative stress and excitotoxicity, both important phenomena in the pathogenesis of Parkinson's disease (PD). We used mice lacking xCT (xCT(-/-) mice), the specific subunit of system x(c)(-), to investigate the involvement of this antiporter in PD. Although cystine that is imported via system x(c)(-) is reduced to cysteine, the rate-limiting substrate in the synthesis of glutathione, deletion of xCT did not result in decreased glutathione levels in striatum. Accordingly, no signs of increased oxidative stress could be observed in striatum or substantia nigra of xCT(-/-) mice. In sharp contrast to expectations, xCT(-/-) mice were less susceptible to 6-hydroxydopamine (6-OHDA)-induced neurodegeneration in the substantia nigra pars compacta compared to their age-matched wild-type littermates. This reduced sensitivity to a PD-inducing toxin might be related to the decrease of 70% in striatal extracellular glutamate levels that was observed in mice lacking xCT. The current data point toward system x(c)(-) as a possible target for the development of new pharmacotherapies for the treatment of PD and emphasize the need to continue the search for specific ligands for system x(c)(-).
Assuntos
Sistema y+ de Transporte de Aminoácidos/deficiência , Dopamina/fisiologia , Neurônios/efeitos dos fármacos , Oxidopamina/toxicidade , Envelhecimento/fisiologia , Sistema y+ de Transporte de Aminoácidos/fisiologia , Animais , Corpo Estriado/metabolismo , Cistina/metabolismo , Glioma/metabolismo , Ácido Glutâmico/metabolismo , Glutationa/metabolismo , Camundongos , Camundongos Knockout , Doenças Neurodegenerativas/prevenção & controle , Neurônios/metabolismo , Substância Negra/metabolismo , Células Tumorais CultivadasRESUMO
Neuropeptides appear to be of importance when the central nervous system (CNS) is challenged, such as during high-frequency firing and pathologic conditions. Potential advantages of treatments that target neuropeptide systems in comparison to classical neurotransmitter systems and ion channels revolve around the subject of efficacy as well as the reduced likelihood of side effects, thus making them attractive candidates for the development of new clinical applications for various disorders. The number of neuropeptides linked to epilepsy is on the rise, reflecting the increased interest of researchers in this domain. Ghrelin has only very recently been introduced into the field of epilepsy, and has already led to contradictory clinical publications. There is a great paucity with regard to what mechanism of action is utilized by ghrelin to inhibit seizures. In this review we disclose how we can better understand the mechanism ghrelin uses to prevent seizures, which indirectly could give an insight to researchers who are studying ghrelin in other fields of research.
Assuntos
Anticonvulsivantes/uso terapêutico , Epilepsia/tratamento farmacológico , Grelina/uso terapêutico , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/fisiologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Epilepsia/patologia , Humanos , Camundongos , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Neuropeptídeos/uso terapêutico , Receptores de Grelina/genética , Receptores de Grelina/metabolismoRESUMO
Vagus nerve stimulation (VNS) is an effective adjunctive treatment for medically refractory epilepsy. In this study, we measured VNS-induced changes in hippocampal neurotransmitter levels and determined their potential involvement in the anticonvulsive action of VNS, to elucidate the mechanism of action responsible for the seizure suppressing effect of VNS in an animal model for limbic seizures. We used in vivo intracerebral microdialysis to measure VNS-induced changes in hippocampal extracellular concentrations of noradrenaline, dopamine, serotonin and GABA in freely moving, male Wistar rats. During the same experiment, the effect of VNS on pilocarpine-induced limbic seizures was assessed using video-EEG monitoring. The involvement of VNS-induced increases in hippocampal noradrenaline in the mechanims of action of VNS was evaluated by blocking hippocampal α(2)-receptors. VNS produced a significant increase in hippocampal noradrenaline concentration (69 ± 16% above baseline levels). VNS also increased the latency between pilocarpine infusion and the onset of epileptiform discharges, and reduced the duration and severity of pilocarpine-induced limbic seizures. A strong positive correlation was found between the noradrenergic and anticonvulsive effects of VNS. Blockade of hippocampal α(2 -receptors reversed the seizure-suppressing effect of VNS. VNS induces increases in extracellular hippocampal noradrenaline, which are at least partly responsible for its seizure-suppressing effect in a model for limbic seizures, and constitute a potential biomarker for the efficacy of VNS in temporal lobe epilepsy.
Assuntos
Hipocampo/metabolismo , Norepinefrina/metabolismo , Convulsões/patologia , Convulsões/terapia , Estimulação do Nervo Vago/métodos , Antagonistas Adrenérgicos alfa/farmacologia , Antagonistas Adrenérgicos alfa/uso terapêutico , Animais , Benzazepinas/farmacologia , Benzazepinas/uso terapêutico , Modelos Animais de Doenças , Eletroencefalografia , Hipocampo/efeitos dos fármacos , Masculino , Microdiálise/métodos , Agonistas Muscarínicos/efeitos adversos , Pilocarpina/efeitos adversos , Ratos , Ratos Wistar , Convulsões/induzido quimicamente , Estatística como Assunto , Gravação em Vídeo/métodosRESUMO
Several studies have revealed that manipulation of the renin angiotensin system results in reduced progression of nigrostriatal damage in different animal models of Parkinson's disease. In the present work, the effect of daily treatment of rats with the angiotensin II (Ang II) type 1 (AT(1) ) receptor antagonist candesartan (3 mg/kg per day, s.c.) initiated 7 days before the intrastriatal injection of 6-hydroxydopamine (6-OHDA) was investigated by means of tyrosine hydroxylase-positive cell counts in the substantia nigra, and dopamine and 3,4-dihydroxyphenylacetic acid measurements in the striatum. In this experimental set-up, candesartan protected dopaminergic neurons of the nigrostriatal tract against the neurotoxin-induced cell death. However, the beneficial effects of AT(1) receptor blockade were not confirmed when treatment was started 24 h after the lesion, suggesting that candesartan interferes with the early events of the 6-OHDA-induced cell death. Stimulation of the AT(1) receptor with Ang II increased the formation of hydroxyl radicals in the striatum of intact rats as measured by the in vivo microdialysis salicylate trapping technique. This Ang II-induced production of reactive oxygen species was suppressed by candesartan perfusion. Furthermore, the Ang II-induced production of reactive oxygen species was nicotinamide adenine dinucleotide phosphate - oxidase and protein kinase C dependent as it could be blocked in the presence of apocynin, an nicotinamide adenine dinucleotide phosphate - oxidase inhibitor, and chelerythrine, an inhibitor of protein kinase C. Together, these data further support the hypothesis that Ang II might contribute in an early stage to the neurotoxicity of 6-OHDA by reinforcing the cascade of oxidative stress.
Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Benzimidazóis/farmacologia , Neurônios Dopaminérgicos/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Oxidopamina/farmacologia , Tetrazóis/farmacologia , Animais , Compostos de Bifenilo , Contagem de Células , Morte Celular/efeitos dos fármacos , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Corpo Estriado/patologia , Dopamina/metabolismo , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Masculino , Microdiálise , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Substância Negra/efeitos dos fármacos , Substância Negra/metabolismo , Substância Negra/patologiaRESUMO
BACKGROUND: Stroke is an important cause of morbidity and mortality and few therapies exist thus far. Mild hypothermia (33°C) is a promising neuroprotective strategy to improve outcome after ischemic stroke. However, its complete mechanism of action has not yet been fully elaborated. This study is the first to investigate whether this neuroprotection occurs through modulation of the neuroinflammatory response after stroke in a time-dependent manner. METHODS: The Endothelin-1 (Et-1) model was used to elicit a transient focal cerebral ischemia in male Wistar rats. In this model, the core and penumbra of the insult are represented by the striatum and the cortex respectively. We assessed the effects of 2 hours of hypothermia, started 20 minutes after Et-1 injection on neurological outcome and infarct volume. Furthermore, pro- and anti-inflammatory cytokine expression was determined using ELISA. Microgliosis and astrogliosis were investigated using CD-68 and GFAP staining respectively. All parameters were determined 8, 24, 72 hours and 1 week after the administration of Et-1. RESULTS: Et-1 infusion caused neurological deficit and a reproducible infarct size which increased up to 3 days after the insult. Both parameters were significantly reduced by hypothermia. The strongest reduction in infarct volume with hypothermia, at 3 days, corresponded with increased microglial activation. Reducing the brain temperature affected the stroke induced increase in interleukin-1ß and tumor necrosis factor α in the striatum, 8 hours after its induction, but not at later time points. Transforming growth factor ß increased as a function of time after the Et-1-induced insult and was not influenced by cooling. Hypothermia reduced astrogliosis at 1 and 3 days after stroke onset. CONCLUSIONS: The beneficial effects of hypothermia after stroke on infarct volume and functional outcome coincide with a time-dependent modulation of the cytokine expression and gliosis.
Assuntos
Citocinas/metabolismo , Endotelina-1/farmacologia , Gliose , Hipotermia/metabolismo , Ataque Isquêmico Transitório/induzido quimicamente , Ataque Isquêmico Transitório/metabolismo , Ataque Isquêmico Transitório/patologia , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Gliose/induzido quimicamente , Gliose/metabolismo , Gliose/patologia , Interleucina-1beta/metabolismo , Ataque Isquêmico Transitório/fisiopatologia , Masculino , Ratos , Ratos Wistar , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
In the past, antidepressants have been thought to possess proconvulsant properties. This assumption remains controversial, however, because anticonvulsant effects have been attributed to certain antidepressants. To date, it remains unclear which antidepressants can be used for the treatment of patients with epilepsy with depression. The present study was designed to determine the anticonvulsant and/or proconvulsant effects of three antidepressants (citalopram, reboxetine, bupropion) against pilocarpine- and pentylenetetrazole-induced acute seizures in larval zebrafish and mice. In zebrafish, all antidepressants were anticonvulsant in the pentylenetetrazole model. In addition, citalopram was anticonvulsant in the zebrafish pilocarpine model, whereas reboxetine and bupropion were without significant effect. In mice all three antidepressants increased some thresholds for pentylenetetrazole-induced convulsive-like behaviors at varying doses, whereas thresholds for pilocarpine-induced convulsive-like behaviors were generally lowered, particularly at the highest doses tested. In general we conclude that the convulsant liability of antidepressants is model and concentration dependent.
Assuntos
Antidepressivos/uso terapêutico , Convulsivantes/toxicidade , Convulsões/induzido quimicamente , Convulsões/tratamento farmacológico , Análise de Variância , Animais , Animais Geneticamente Modificados , Comportamento Animal/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Interações Medicamentosas , Proteínas de Fluorescência Verde/genética , Masculino , Camundongos , Pentilenotetrazol/toxicidade , Pilocarpina/toxicidade , Convulsões/mortalidade , Convulsões/fisiopatologia , Peixe-ZebraRESUMO
Noradrenaline exerts inhibitory effects on seizure susceptibility. Subtype selective agonists and antagonists were used to identify the anticonvulsant hippocampal adrenoreceptors. Intrahippocampal dialysis was used for administration of all compounds, including pilocarpine for limbic seizure induction, and as the neurotransmitter sampling tool. The noradrenaline reuptake inhibitor maprotiline mediated anticonvulsant effects, associated with dose-dependent increases in extracellular hippocampal noradrenaline, dopamine and GABA levels. At high concentrations, maprotiline produced proconvulsant effects associated with high levels of noradrenaline, dopamine and glutamate. Maprotiline's anticonvulsant effect was blocked by administration of either a selective α(2) - and ß(2) -antagonist. α(2) -Antagonist administration with maprotiline was associated with a further increase in noradrenaline and dopamine from maprotiline alone; whereas ß(2) -antagonist administered with maprotiline inhibited the dopamine increases produced by maprotiline. α(1A) -Antagonism blocked the GABA-ergic but not the anticonvulsive effect of maprotiline. These results were confirmed as combined but not separate α(2) - and ß(2) -adrenoreceptor stimulation, using selective agonists, inhibited limbic seizures. Interestingly, α(1A) -receptor stimulation and α(1D) -antagonism alone also inhibited seizures associated with respectively significant hippocampal GABA increases and glutamate decreases. The main findings of this study are that (i) increased hippocampal noradrenergic neurotransmission inhibits limbic seizures via combined α(2) - and ß(2) -receptor activation and (ii) α(1A) - and α(1D) -adrenoreceptors mediate opposite effects on hippocampal excitability.
Assuntos
Agonistas Adrenérgicos/farmacologia , Antagonistas Adrenérgicos/farmacologia , Hipocampo/metabolismo , Sistema Límbico/metabolismo , Receptores Adrenérgicos/metabolismo , Convulsões/metabolismo , Agonistas Adrenérgicos/uso terapêutico , Antagonistas Adrenérgicos/uso terapêutico , Animais , Hipocampo/química , Hipocampo/efeitos dos fármacos , Sistema Límbico/química , Sistema Límbico/efeitos dos fármacos , Masculino , Subunidades Proteicas/agonistas , Subunidades Proteicas/antagonistas & inibidores , Subunidades Proteicas/metabolismo , Ratos , Ratos Wistar , Receptores Adrenérgicos alfa 1/metabolismo , Receptores Adrenérgicos alfa 2/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Convulsões/tratamento farmacológicoRESUMO
Intracerebroventricular (i.c.v.) administration of angiotensin IV (Ang IV) or Leu-Val-Val-haemorphin 7 (LVV-H7) improves memory performance in normal rats and reverses memory deficits in rat models for cognitive impairment. These memory effects were believed to be mediated via the putative 'AT4 receptor'. However, this binding site was identified as insulin-regulated aminopeptidase (IRAP). Correspondingly, Ang IV and LVV-H7 were characterised as IRAP inhibitors. This study investigates whether and how IRAP may be involved in the central effects of Ang IV and LVV-H7. We determined the effects of i.c.v. administration of Ang IV or LVV-H7 on hippocampal neurotransmitter levels using microdialysis in rats. We observed that Ang IV modulates hippocampal acetylcholine levels, whereas LVV-H7 does not. This discrepancy was reflected in the observation that Ang IV binds with micromolar affinity to the AT1 receptor whereas no binding affinity was observed for LVV-H7. Correspondingly, we demonstrated that the AT1 receptor is involved in the effects of Ang IV on hippocampal neurotransmitter levels and on spatial working memory in a plus maze spontaneous alternation task. However, the AT1 receptor was not involved in the spatial memory facilitating effect of LVV-H7. Finally, we demonstrated that Ang IV did not diffuse to the hippocampus following i.c.v. injection, suggesting an extrahippocampal site of action. We propose that AT1 receptors are implicated in the neurochemical and cognitive effects of Ang IV, whereas LVV-H7 may mediate its effects via IRAP.
Assuntos
Angiotensina II/análogos & derivados , Hemoglobinas/administração & dosagem , Hipocampo/efeitos dos fármacos , Memória de Curto Prazo/efeitos dos fármacos , Neurotransmissores/metabolismo , Fragmentos de Peptídeos/administração & dosagem , Receptor Tipo 1 de Angiotensina/fisiologia , Angiotensina II/administração & dosagem , Angiotensina II/farmacocinética , Animais , Relação Dose-Resposta a Droga , Hipocampo/diagnóstico por imagem , Hipocampo/metabolismo , Injeções Intraventriculares/métodos , Isótopos de Iodo/farmacocinética , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Microdiálise/métodos , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Wistar , Percepção Espacial/efeitos dos fármacos , Distribuição Tecidual/efeitos dos fármacos , Tomografia Computadorizada de Emissão de Fóton Único/métodosRESUMO
Neuroinflammation is a key element in the ischemic cascade after cerebral ischemia that results in cell damage and death in the subacute phase. However, anti-inflammatory drugs do not improve outcome in clinical settings suggesting that the neuroinflammatory response after an ischemic stroke is not entirely detrimental. This review describes the different key players in neuroinflammation and their possible detrimental and protective effects in stroke. Because of its inhibitory influence on several pathways of the ischemic cascade, hypothermia has been introduced as a promising neuroprotective strategy. This review also discusses the influence of hypothermia on the neuroinflammatory response. We conclude that hypothermia exerts both stimulating and inhibiting effects on different aspects of neuroinflammation and hypothesize that these effects are key to neuroprotection.
Assuntos
Isquemia Encefálica/patologia , Hipotermia Induzida , Inflamação/patologia , Inflamação/terapia , Acidente Vascular Cerebral/patologia , Animais , Anti-Inflamatórios/uso terapêutico , Isquemia Encefálica/complicações , Isquemia Encefálica/imunologia , Isquemia Encefálica/fisiopatologia , Quimiocinas/imunologia , Citocinas/imunologia , Proteínas HMGB/imunologia , Humanos , Inflamação/etiologia , Inflamação/imunologia , Integrinas/imunologia , Metaloproteinases da Matriz/imunologia , Fármacos Neuroprotetores/uso terapêutico , Espécies Reativas de Oxigênio/imunologia , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/imunologia , Acidente Vascular Cerebral/fisiopatologiaRESUMO
BACKGROUND AND PURPOSE: Insulin-like growth factor I (IGF-I) exerts neuroprotective effects in both white and gray matter under different detrimental conditions. The purpose of this review is to collect the evidence whether IGF-I is a candidate neuroprotective drug in patients with acute ischemic stroke. RESULTS: IGF-I was found to be neuroprotective in animal models of focal brain ischemia when given >or=2 hours after the insult. Different routes of administration (eg, cerebroventricular, intravenous, and intranasal) were found to be effective. In addition to inhibition of apoptosis and reduction of the infarct volume, IGF-I also improved neurological outcome. Furthermore, there are strong indications that IGF-I can also stimulate the regeneration of neural tissue. CONCLUSIONS: Additional studies are required to reveal the neuroprotective mechanisms of IGF-I in detail and to elucidate the role of IGF-binding proteins. Preclinical studies in relevant animal models for studying stroke (ie, hypertensive, diabetic, or aged animals) should be done testing different doses and routes of IGF-I administration and different combinations of IGF-I and IGF-binding proteins.
Assuntos
Isquemia Encefálica/tratamento farmacológico , Fator de Crescimento Insulin-Like I/farmacologia , Fármacos Neuroprotetores/farmacologia , Acidente Vascular Cerebral/tratamento farmacológico , Doença Aguda , Animais , Modelos Animais de Doenças , HumanosRESUMO
BACKGROUND: Efficient protection of dopaminergic neurons against a subsequent 6-hydroxydopamine lesion by glial cell line-derived neurotrophic factor (GDNF) gene delivery has been demonstrated. By contrast, the neurorestorative effects of GDNF administered several weeks after the toxin have been less characterized. In particular, whether these were permanent or dependent on the continuous presence of GDNF remains elusive. METHODS: A tetracycline-inducible adeno-associated virus (AAV)-1 vector expressing human GDNF cDNA was administered unilaterally in the rat striatum 5 weeks after 6-hydroxydopamine. Rats were treated with doxycycline (dox) or untreated from the day of vector injection until sacrifice (4 or 14 weeks). A sub-group was dox-treated for 7 weeks then untreated until 14 weeks. The motor behavior was assessed by amphetamine-induced rotations and spontaneous forelimb asymmetry. The amounts of tyrosine hydroxylase (TH), serine-40-phosphorylated TH (S40-TH) and aromatic amino acid decarboxylase (AADC) proteins were compared by western blotting and the dopamine levels quantified by high-performance liquid chromatography. RESULTS: Dox-dependent behavioral improvements were demonstrated 4 weeks post-vector injection. At later time points, spontaneous partial recovery was observed in all rats, but no further improvement was found in dox-treated animals. TH levels were significantly increased in dox-treated rats at all time points. By contrast, striatal dopamine and S40-TH were increased at 4 weeks, but not 14 weeks, and AADC remained unchanged. Dox withdrawal after 7 weeks, resulted in TH levels comparable to the controls at 14 weeks. CONCLUSIONS: Delayed GDNF gene delivery only transiently improved dopaminergic function. Over the long term, TH was more abundant, but not functional, and the increase was lost when GDNF gene expression was switched off.
Assuntos
Fator Neurotrófico Derivado de Linhagem de Célula Glial/administração & dosagem , Doença de Parkinson Secundária/metabolismo , Doença de Parkinson Secundária/terapia , Adrenérgicos/administração & dosagem , Adrenérgicos/efeitos adversos , Animais , Dependovirus/genética , Dopamina/análise , Dopamina/biossíntese , Doxiciclina/administração & dosagem , Feminino , Expressão Gênica/efeitos dos fármacos , Técnicas de Transferência de Genes , Vetores Genéticos/administração & dosagem , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/biossíntese , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Humanos , Atividade Motora/efeitos dos fármacos , Oxidopamina/administração & dosagem , Oxidopamina/efeitos adversos , Doença de Parkinson Secundária/induzido quimicamente , Ratos , Fatores de Tempo , Tirosina 3-Mono-Oxigenase/análise , Tirosina 3-Mono-Oxigenase/biossínteseRESUMO
The IRAP ligands Angiotensin IV (Ang IV) and LVV-haemorphin 7 (LVV-H7) enhance performance in a range of memory paradigms in normal rats and ameliorate memory deficits in rat models for amnesia. The mechanism by which these peptides facilitate memory remains to be elucidated. In recent in vitro experiments, we demonstrated that Ang IV and LVV-H7 potentiate activity-evoked glucose uptake into hippocampal neurons. This raises the possibility that IRAP ligands may facilitate memory in hippocampus-dependent tasks through enhancement of hippocampal glucose uptake. Acute intracerebroventricular (i.c.v.) administration of 1nmol Ang IV or 0.1nmol LVV-H7 in 3 months-old Sprague-Dawley rats enhanced spatial working memory in the plus maze spontaneous alternation task. Extracellular hippocampal glucose levels were monitored before, during and after behavioral testing using in vivo microdialysis. Extracellular hippocampal glucose levels decreased significantly to about 70% of baseline when the animals explored the plus maze, but remained constant when the animals were placed into a novel control chamber. Ang IV and LVV-H7 did not significantly alter hippocampal glucose levels compared to control animals in the plus maze or control chamber. Both peptides had no effect on hippocampal blood flow as determined by laser Doppler flowmetry, excluding that either peptide increased the hippocampal supply of glucose. We demonstrated for the first time that Ang IV and LVV-H7 enhance spatial working memory in the plus maze spontaneous alternation task but no in vivo evidence was found for enhanced hippocampal glucose uptake or blood flow.
Assuntos
Angiotensina II/análogos & derivados , Hemoglobinas/farmacologia , Hipocampo/efeitos dos fármacos , Memória/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Percepção Espacial/efeitos dos fármacos , Vasoconstritores/farmacologia , Angiotensina II/farmacologia , Animais , Cateterismo , Circulação Cerebrovascular/efeitos dos fármacos , Espaço Extracelular/metabolismo , Fluxômetros , Glucose/metabolismo , Hipocampo/irrigação sanguínea , Hipocampo/metabolismo , Injeções Intraventriculares , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/fisiologia , Microdiálise , Atividade Motora/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Percepção Espacial/fisiologia , Comportamento Espacial/efeitos dos fármacos , Comportamento Espacial/fisiologiaRESUMO
The neurobiological relationships between epilepsy and depression are receiving increased experimental attention. A key role for limbic monoamines in depression has been established and we recently showed the importance of hippocampal monoamines in limbic seizure control. We here studied whether anticonvulsant compounds are antidepressant and can elevate hippocampal dopamine (DA) or serotonin (5-HT) levels determined by in vivo microdialysis in rats. We used assessment of seizure severity in the focal pilocarpine model, antidepressant-like activity within the rat forced swim and the mouse tail suspension tests, and locomotor activity in an open field as behavioural tests. We studied the tricyclic antidepressant imipramine, the selective 5-HT reuptake inhibitor citalopram and the selective DA reuptake blocker GBR-12909. These compounds with combined antidepressant-anticonvulsant properties all directly enhanced extracellular hippocampal DA or 5-HT levels. Since glutamate-mediated hyperexcitability in temporal lobe regions seems to be involved in disturbed emotional behaviour, we next investigated possible antidepressant effects and hippocampal DA or 5-HT modulations exerted by selective ionotropic and metabotropic glutamate receptor ligands with anticonvulsant properties. Combined anticonvulsant-antidepressant activities of the NMDA antagonist MK-801 and the mGluR group I antagonists (AIDA, MPEP) were also associated with locally elicited increases in hippocampal DA and/or 5-HT levels. This study highlights that the hippocampus is an important site of action of combined anticonvulsant-antidepressant and monoamine enhancing effects.
Assuntos
Anticonvulsivantes/farmacologia , Antidepressivos/farmacologia , Dopamina/metabolismo , Hipocampo/metabolismo , Serotonina/metabolismo , Animais , Antidepressivos Tricíclicos/farmacologia , Cromatografia Líquida , Citalopram/farmacologia , Inibidores da Captação de Dopamina/farmacologia , Antagonistas GABAérgicos/farmacologia , Elevação dos Membros Posteriores , Hipocampo/efeitos dos fármacos , Indicadores e Reagentes , Masculino , Camundongos , Microdiálise , Atividade Motora/efeitos dos fármacos , Piperazinas/farmacologia , Ratos , Ratos Wistar , Receptores de Glutamato/efeitos dos fármacos , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Convulsões/tratamento farmacológico , Convulsões/fisiopatologia , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Natação/psicologiaRESUMO
Microdialysis in intact and denervated striatum of unilaterally 6-hydroxydopamine (6-OHDA) lesioned rats was used to investigate whether CR 3394, N-[2-(3,5-dimethyl-1-adamantyl)ethyl]acetamidine, an adamantane derivative with preferential selectivity for the NR2B subunit of the NMDA receptor, has dopamine releasing properties in vivo. We also investigated whether this NMDA antagonist can potentiate the effects of L-Dopa on extracellular dopamine in these animals. After systemic injection, there was no significant effect of CR 3394 on extracellular dopamine, at all doses studied (1, 5 and 20 mg/kg i.p.), in either intact or in denervated striatum. On the other hand, striatal perfusion with 100 microM of the compound elicited release of dopamine in intact, but not in denervated striatum. In denervated striatum of the 6-OHDA-lesioned rats, CR 3394 (5 mg/kg) significantly enhanced the dopamine release induced by L-Dopa administration (25 mg/kg i.p.) in combination with benserazide (10 mg/kg i.p.). In particular, the onset of action of L-Dopa was potentiated. However, when combined with a subthreshold dose of L-Dopa (5 mg/kg), the effects of CR 3394 were lost. We conclude that CR 3394, like other NR2B receptor antagonists, has dopamine releasing properties in vivo. It enhances the effects of suprathreshold doses of L-Dopa in the denervated striatum, but not of low doses of L-Dopa. Therefore, future studies are necessary to establish the potential of selective NR2B receptor antagonists as L-Dopa-sparing agents.