RESUMEN
Amyloid-ß oligomers (Aßo) are the most pathologically relevant Aß species in Alzheimer's disease (AD), because they induce early synaptic dysfunction that leads to learning and memory impairments. In contrast, increasing VEGF (Vascular Endothelial Growth Factor) brain levels have been shown to improve learning and memory processes, and to alleviate Aß-mediated synapse dysfunction. Here, we designed a new peptide, the blocking peptide (BP), which is derived from an Aßo-targeted domain of the VEGF protein, and investigated its effect on Aß-associated toxicity. Using a combination of biochemical, 3D and ultrastructural imaging, and electrophysiological approaches, we demonstrated that BP strongly interacts with Aßo and blocks Aß fibrillar aggregation process, leading to the formation of Aß amorphous aggregates. BP further impedes the formation of structured Aßo and prevents their pathogenic binding to synapses. Importantly, acute BP treatment successfully rescues long-term potentiation (LTP) in the APP/PS1 mouse model of AD, at an age when LTP is highly impaired in hippocampal slices. Moreover, BP is also able to block the interaction between Aßo and VEGF, which suggests a dual mechanism aimed at both trapping Aßo and releasing VEGF to alleviate Aßo-induced synaptic damage. Our findings provide evidence for a neutralizing effect of the BP on Aß aggregation process and pathogenic action, highlighting a potential new therapeutic strategy.
Asunto(s)
Enfermedad de Alzheimer , Péptidos beta-Amiloides , Ratones , Animales , Péptidos beta-Amiloides/toxicidad , Péptidos beta-Amiloides/metabolismo , Factor A de Crecimiento Endotelial Vascular/farmacología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Enfermedad de Alzheimer/metabolismo , Memoria/fisiología , Aprendizaje , Sinapsis/metabolismo , Fragmentos de Péptidos/farmacología , Fragmentos de Péptidos/metabolismoRESUMEN
BACKGROUND: The introduction of viral transneuronal tracers in the toolbox of neural tract-tracing methods has been an important addition in the field of connectomics for deciphering circuit-level architecture of the nervous system. One of the added values of viral compared to conventional retrograde tracers, in particular of rabies virus, is to provide a Golgi staining-like view of the infected neurons, revealing the thin dendritic arborizations and the spines that are major post-synaptic seats of neuronal connections. NEWMETHOD: Here, we comparatively illustrate the characteristics of the labeling obtained in the same model system, the basal ganglia circuitry, by different retrograde viral tracing approaches, using the Bartha strain of pseudorabies virus, the SAD and CVS strains of rabies virus and by the conventional retrograde tracer cholera toxin B. To best contrast the differences in the capacity of these tracers to reveal the dendritic morphology in details, we focused on one population of first-order infected neurons in the striatum, which exhibit high spine density, after tracer injection in the substantia nigra. RESULTS AND CONCLUSION: None of the viruses tested allowed to detect as many neurons as with cholera toxin B, but the SAD and CVS strains of rabies virus had the advantage of enabling detailed Golgi-like visualisation of the dendritic trees, the best numerical detection being offered by the transneuronal rCVS-N2c-P-mCherry while poor labeling was provided by rCVS-N2c-M-GFP. Results also suggest that, besides different viral properties, technical issues about constructs and detection methods contribute to apparently different efficiencies among the viral approaches.
Asunto(s)
Herpesvirus Suido 1 , Virus de la Rabia , Animales , Encéfalo , Neuronas , Coloración y EtiquetadoRESUMEN
INTRODUCTION: Indoor microbial toxicity is suspected to cause some building-related symptoms, but supporting epidemiological data are lacking. OBJECTIVE: We examined whether the in vitro toxicity of indoor samples from school buildings was associated with work-related health symptoms (building-related symptoms, BRS). METHODS: Administrators of the Helsinki City Real Estate Department selected 15 schools for the study, and a questionnaire on symptoms connected to work was sent to the teachers in the selected schools for voluntary completion. The cellular toxicity of classroom samples was determined by testing substances extracted from wiped indoor dust and by testing microbial biomass that was cultured on fallout plates. Boar sperm cells were used as indicator cells, and motility loss was the indicator for toxic effects. The effects were expressed as the half maximal effective concentration (EC50) at which >50% of the exposed boar sperm cells were immobile compared to vehicle control. RESULTS: Completed symptom questionnaires were received from 232 teachers [median age, 43 years; 190 (82.3%) women] with a median time of 6 years working at their school. Samples from their classrooms were available and were assessed for cellular toxicity. The Poisson regression model showed that the impact of extracts of surface-wiped school classroom dust on teacher work-related BRS was 2.8-fold (95% CI: 1.6-4.9) higher in classrooms with a toxic threshold EC50 of 6µgml-1 versus classrooms with insignificant EC50 values (EC50 >50µgml-1); P<0.001. The number of symptoms that were alleviated during vacation was higher in school classrooms with high sperm toxicity compared to less toxic sites; the RR was 1.9 (95% CI: 1.1-3.3, P=0.03) for wiped dust extracts. CONCLUSIONS: Teachers working in classrooms where the samples showed high sperm toxicity had more BRS. The boar sperm cell motility inhibition assay appears promising as a tool for demonstrating the presence of indoor substances associated with BRS.
Asunto(s)
Contaminación del Aire Interior/efectos adversos , Exposición Profesional/efectos adversos , Maestros/estadística & datos numéricos , Instituciones Académicas/estadística & datos numéricos , Síndrome del Edificio Enfermo , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Adulto , Microbiología del Aire , Contaminación del Aire Interior/análisis , Animales , Estudios Transversales , Polvo/análisis , Monitoreo del Ambiente , Femenino , Finlandia , Humanos , Masculino , Persona de Mediana Edad , Exposición Profesional/análisis , Síndrome del Edificio Enfermo/epidemiología , PorcinosRESUMEN
Vascular endothelial growth factor (VEGF) is known to be required for the action of antidepressant therapies but its impact on brain synaptic function is poorly characterized. Using a combination of electrophysiological, single-molecule imaging and conditional transgenic approaches, we identified the molecular basis of the VEGF effect on synaptic transmission and plasticity. VEGF increases the postsynaptic responses mediated by the N-methyl-D-aspartate type of glutamate receptors (GluNRs) in hippocampal neurons. This is concurrent with the formation of new synapses and with the synaptic recruitment of GluNR expressing the GluN2B subunit (GluNR-2B). VEGF induces a rapid redistribution of GluNR-2B at synaptic sites by increasing the surface dynamics of these receptors within the membrane. Consistently, silencing the expression of the VEGF receptor 2 (VEGFR2) in neural cells impairs hippocampal-dependent synaptic plasticity and consolidation of emotional memory. These findings demonstrated the direct implication of VEGF signaling in neurons via VEGFR2 in proper synaptic function. They highlight the potential of VEGF as a key regulator of GluNR synaptic function and suggest a role for VEGF in new therapeutic approaches targeting GluNR in depression.
Asunto(s)
Factor A de Crecimiento Endotelial Vascular/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Animales , Conducta Animal , Técnicas de Cultivo de Célula , Potenciales Postsinápticos Excitadores , Miedo , Hipocampo/metabolismo , Ratones , Plasticidad Neuronal/fisiología , Neuronas/metabolismo , Subunidades de Proteína , Receptores de Glutamato/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapsis/metabolismo , Transmisión Sináptica/fisiología , Factor A de Crecimiento Endotelial Vascular/fisiología , Receptor 2 de Factores de Crecimiento Endotelial Vascular/fisiologíaRESUMEN
Activation of group III metabotropic glutamate (mGlu) receptors has been recently highlighted as a potential approach in the treatment of Parkinson's disease (PD). This study evaluates the antiparkinsonian action of systemic administration of the broad-spectrum agonist of group III mGlu receptors, 1-aminocyclopentane-1,3,4-tricarboxylic acid (ACPT-I), and investigates its site of action within the basal ganglia circuitry. Acute injection of ACPT-I reverses haloperidol-induced catalepsy, an index of akinesia in rodents. In a rat model of early PD based on partial bilateral nigrostriatal lesions, chronic (2weeks) administration of ACPT-I is required to efficiently alleviate the akinetic deficit evidenced in a reaction time task. This treatment counteracts the post-lesional increases in the gene expression of cytochrome oxidase subunit I, a metabolic marker of neuronal activity, in the overall subthalamic nucleus and in the lateral motor part of the substantia nigra pars reticulata (SNr) but has no effect in the globus pallidus. Paradoxically, ACPT-I administration in sham animals impairs performance and induces overexpression of cytochrome oxidase subunit I mRNA in the lateral SNr, and has no effect in the subthalamic nucleus or globus pallidus. Altogether, our results provide new evidence for the antiparkinsonian efficiency of group III mGlu receptor agonism, point to the regulation of the overactive subthalamo-nigral connection as a main site of action in an early stage of PD and underline the complex interplay between these receptors and the dopaminergic system to regulate basal ganglia function in control and PD conditions.
Asunto(s)
Agonistas de Aminoácidos Excitadores/farmacología , Trastornos Parkinsonianos/tratamiento farmacológico , Receptores de Glutamato Metabotrópico/agonistas , Sustancia Negra/efectos de los fármacos , Núcleo Subtalámico/efectos de los fármacos , Animales , Ciclopentanos/farmacología , Modelos Animales de Enfermedad , Masculino , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/metabolismo , Trastornos Parkinsonianos/metabolismo , Ratas , Ratas Wistar , Receptores de Glutamato Metabotrópico/metabolismo , Estereoisomerismo , Sustancia Negra/metabolismo , Núcleo Subtalámico/metabolismo , Ácidos Tricarboxílicos/farmacologíaRESUMEN
The presence, quantity and origins of potentially toxic airborne substances were searched in moisture damaged indoor environments, where building related ill health symptoms were suspected and reference sites with no health complaints. Boar spermatozoa were used as the toxicity sensor. Indoor aerosols and dusts were collected from kindergartens, schools, offices and residences (n=25) by electrostatic filtering, vacuuming, wiping from elevated surfaces and from the interior of personal computers. Toxicity was measured from the ethanol or methanol extracts of the dusts and aerosols. EC(50) was expressed as the lowest concentration of the airborne substance that inhibited motility of >50% of the exposed sperm cells compared to vehicle control, within 30 min, 1 day or 3-4 days of exposure. Remarkably toxic aerosols (EC(50) Asunto(s)
Contaminación del Aire Interior/efectos adversos
, Espermatozoides/efectos de los fármacos
, Pruebas de Toxicidad/métodos
, Aerosoles/toxicidad
, Animales
, Técnicas Biosensibles
, Polvo
, Masculino
, Motilidad Espermática/efectos de los fármacos
, Espermatozoides/metabolismo
, Electricidad Estática
, Porcinos
, Agua/efectos adversos
RESUMEN
The types of sodium channels that are expressed by neurons shape the rising phase of action potentials and influence patterns of action potential discharge. With regard to the enteric nervous system (ENS), there is uncertainty about which channels are expressed, and in particular it is unknown whether Na(v)1.7 is present. We designed specific probes for the guinea pig Na(v)1.7 alpha subunit as well as for the other tetrodotoxin (TTX)-sensitive alpha subunits (Na(v)1.1, Na(v)1.2, Na(v)1.3, and Na(v)1.6) in order to perform in situ hybridization (ISH) histochemistry on guinea pig myenteric ganglia. We established that only Na(v)1.7 mRNA and Na(v)1.3 mRNA are expressed in these ganglia. The ISH signal for Na(v)1.7 transcripts was found in seemingly all the myenteric neurons. The expression of the Na(v)1.3 alpha subunit was confirmed by immunohistochemistry in a large proportion (62%) of the myenteric neuron population. This population included enteric sensory neurons. Na(v)1.6 immunoreactivity, absent from myenteric neurons, was detected in glial cells only when a high anti-Na(v)1.6 antibody concentration was used. This suggests that the Na(v)1.6 alpha subunit and mRNA are present only at low levels, which is consistent with the fact that no Na(v)1.6 mRNA could be detected in the ENS by ISH. The fact that adult myenteric neurons are endowed with only two TTX-sensitive alpha subunits, namely, Na(v)1.3 and Na(v)1.7, emphasizes the singularity of the ENS. Both these subunits, known to have slow-inactivation kinetics, are well adapted for generating action potentials from slow excitatory postsynaptic potentials, a mode of synaptic transmission that applies to all ENS neuron types.
Asunto(s)
Sistema Nervioso Entérico/metabolismo , Canales de Sodio/metabolismo , Animales , Encéfalo/citología , Encéfalo/metabolismo , Sistema Nervioso Entérico/citología , Ganglios Espinales/citología , Ganglios Espinales/metabolismo , Cobayas , Inmunohistoquímica , Hibridación in Situ/métodos , Neuronas Aferentes/metabolismo , Sondas de Oligonucleótidos , Subunidades de Proteína/clasificación , Subunidades de Proteína/genética , ARN Mensajero/análisis , Nervio Ciático/citología , Nervio Ciático/metabolismo , Sensibilidad y Especificidad , Canales de Sodio/efectos de los fármacos , Canales de Sodio/genética , Tetrodotoxina , Distribución TisularRESUMEN
In psychiatric and neurological diseases, sleep is often perturbed. Moreover, recent works on humans and animals tend to show that sleep plays a strong role in memory processes. Reciprocally, sleep dynamics following a learning task is modified [Hubert, Nature (London) 02663, 1 (2004), Peigneux, Neuron 44, 535 (2004)]. However, sleep analysis in humans and animals is often limited to the total sleep and wake duration quantification. These two parameters are not fully able to characterize the sleep dynamics. In mammals sleep presents a complex organization with an alternation of slow wave sleep (SWS) and paradoxical sleep (PS) episodes. Moreover, it has been shown recently that these sleep episodes are frequently interrupted by micro-arousal (without awakening). We present here a detailed analysis of the basal sleep properties emerging from the mechanisms underlying the vigilance states alternation in an animal model. These properties present a self-organized critical system signature and reveal the existence of two W, two SWS, and a PS structure exhibiting a criticality as met in sand piles. We propose a theoretical model of the sleep dynamics based on several interacting neuronal populations. This new model of sleep dynamics presents the same properties as experimentally observed, and explains the variability of the collected data. This experimental and theoretical study suggests that sleep dynamics shares several common features with critical systems.
RESUMEN
There is growing experimental evidence for the implication of glutamate-mediated mechanisms both in the pathophysiology of Parkinson's disease and in the development of dyskinesias with long-term administration of L-3,4-dihydroxyphenylalanine (L-DOPA). However, the impact of this treatment on glutamate transmission in the basal ganglia has been poorly investigated. In this study, we examined the effects of 6-hydroxydopamine-induced lesion of nigral dopamine neurons with or without subsequent chronic L-DOPA treatment on several parameters of glutamate system function in the rat striatum and substantia nigra pars reticulata. All the lesioned animals treated with L-DOPA developed severe dyskinesias. Extracellular glutamate levels, measured by microdialysis in freely moving conditions, and gene expression of the glial glutamate transporter GLT1, assessed by in situ hybridization, were unaffected by dopamine lesion or L-DOPA treatment alone, but were both markedly increased on the lesion side of rats with subsequent L-DOPA treatment. No change in the expression of the vesicular glutamate transporters vGluT1 and vGluT2 was measured in striatum. These data show that chronic L-DOPA treatment leading to dyskinesias increases basal levels of glutamate function in basal ganglia. The L-DOPA-induced overexpression of GLT1 may represent a compensatory mechanism involving astrocytes to limit glutamate overactivity and subsequent toxic processes.
Asunto(s)
Ganglios Basales/efectos de los fármacos , Transportador 2 de Aminoácidos Excitadores/biosíntesis , Líquido Extracelular/metabolismo , Ácido Glutámico/biosíntesis , Levodopa/administración & dosificación , Enfermedad de Parkinson/tratamiento farmacológico , Animales , Ganglios Basales/metabolismo , Esquema de Medicación , Líquido Extracelular/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Masculino , Enfermedad de Parkinson/metabolismo , Ratas , Ratas WistarRESUMEN
Microtubules are key cytoskeletal components in the cytoplasm of eukaryotic cells where they have pleiotropic and vital roles in functions such as cell division, trafficking or morphogenesis. Microtubules are especially abundant in neurons. Although microtubules are in many cells dynamic polymers, they exhibit an extreme state of stability in neurons. Previous work has indicated a central role of microtubule associated proteins called STOPs in neuronal microtubule stabilization. We have recently developed STOP null mice. These mice were devoid of stable brain microtubules but to our surprise had nevertheless an apparently normal brain anatomy. However the mice showed synaptic defects affecting different forms of long- and short-term synaptic plasticity. These synaptic defects were associated with severe behavioral defects that showed a remarkable sensitivity to long-term treatment with neuroleptics. We discuss the relationship of the phenotypes observed in STOP null mice with current models of schizophrenia in which the multiple, severe, and neuroleptic sensitive mental disorders caused by the disease are due to a "disease of the synapse".
Asunto(s)
Citoesqueleto/fisiología , Trastornos Mentales/patología , Microtúbulos/fisiología , Neuronas/ultraestructura , Animales , Antipsicóticos/farmacología , Antipsicóticos/uso terapéutico , Modelos Animales de Enfermedad , Humanos , Trastornos Mentales/tratamiento farmacológico , Ratones , Ratones Noqueados , Proteínas Asociadas a Microtúbulos/deficiencia , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/fisiología , Modelos Biológicos , Modelos Neurológicos , Plasticidad Neuronal , Esquizofrenia/tratamiento farmacológico , Esquizofrenia/patología , Tubulina (Proteína)/metabolismoRESUMEN
There is growing evidence that the loss of the nigrostriatal dopaminergic neurones induces an overactivity of the corticostriatal glutamatergic pathway which seems to be central to the physiopathology of parkinsonism. Moreover, glutamatergic mechanisms involving NMDA receptors have been shown to interfere with the therapeutical action of levodopa. Given the key role played by uptake processes in glutamate neurotransmission, this study examined the effects of nigrostriatal deafferentation and of levodopa treatment on the striatal expression of the glutamate transporters GLT1, GLAST and EAAC1 in the rat. No significant changes in striatal mRNA levels of these transporters were detected after either levodopa treatment (100 mg/kg; i.p., twice a day for 21 days) or unilateral lesion of the nigrostriatal pathway by intranigral 6-hydroxydopamine injection. In contrast, animals with the lesion subsequently treated with levodopa showed a selective increase (36%) in GLT1 mRNA levels in the denervated striatum versus controls. These animals also showed increased GLT1 protein expression, as assessed by immunostaining and western blotting. These data provide the first evidence that levodopa therapy may interfere with striatal glutamate transmission through change in expression of the primarily glial glutamate transporter GLT1. We further suggest that levodopa-induced GLT1 overexpression may represent a compensatory mechanism preventing neurotoxic accumulation of endogenous glutamate.
Asunto(s)
Sistema de Transporte de Aminoácidos X-AG/genética , Transportador 2 de Aminoácidos Excitadores/genética , Levodopa/farmacología , Sustancia Negra/metabolismo , Simportadores , Sistema de Transporte de Aminoácidos X-AG/biosíntesis , Animales , Proteínas Portadoras/genética , Desnervación , Dopamina/metabolismo , Dopaminérgicos/farmacología , Encefalinas/genética , Encefalinas/metabolismo , Transportador 1 de Aminoácidos Excitadores , Transportador 2 de Aminoácidos Excitadores/biosíntesis , Transportador 3 de Aminoácidos Excitadores , Femenino , Técnica del Anticuerpo Fluorescente , Proteínas de Transporte de Glutamato en la Membrana Plasmática , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Radiografía , Ratas , Sustancia Negra/diagnóstico por imagen , Sustancia Negra/efectos de los fármacosRESUMEN
It has been shown recently that in mitral cells of the rat olfactory bulb, N-methyl-D-aspartate (NMDA) autoreceptors are activated during mitral cell firing. Here we consider in more details the mechanisms of mitral cell self-excitation and its physiological relevance. We show that both ionotropic NMDA and non-NMDA autoreceptors are activated by glutamate released from primary and secondary dendrites. In contrast to non-NMDA autoreceptors, NMDA autoreceptors are almost exclusively located on secondary dendrites and their activation generates a large and sustained self-excitation. Both intracellularly evoked and miniature NMDA-R mediated synaptic potentials are blocked by intracellular bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) and result from a calcium-dependent release of glutamate. Self-excitation can be produced by a single spike, and trains of spikes result in frequency facilitation. Thus activation of excitatory autoreceptors is a major function of action potentials backpropagating in mitral cell dendrites, which results in an immediate positive feedback counteracting recurrent inhibition and increasing the signal-to-noise ratio of olfactory inputs.
Asunto(s)
Autorreceptores/metabolismo , Dendritas/metabolismo , Ácido Glutámico/metabolismo , Bulbo Olfatorio/metabolismo , Transducción de Señal/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Señalización del Calcio/fisiología , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Antagonistas de Receptores de GABA-A , Técnicas In Vitro , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Bulbo Olfatorio/citología , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , Receptores AMPA/antagonistas & inhibidores , Receptores AMPA/metabolismo , Receptores de Glutamato Metabotrópico/antagonistas & inhibidores , Receptores de Glutamato Metabotrópico/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Receptores de N-Metil-D-Aspartato/metabolismo , Sodio/metabolismoRESUMEN
Recent pathophysiological models of basal ganglia function in Parkinson's disease predict that specific neurochemical changes in the indirect pathway would follow the lack of stimulation of D(2) dopamine receptors. Post mortem studies of the basal ganglia in genetically modified mice lacking functional copies of the D(2) dopamine receptor gene allowed us to test these predictions. When compared with their congenic N(5) wild-type siblings, mice lacking D(2) receptors show an increased expression of enkephalin messenger RNA in the striatum, and an increased activity and expression of cytochrome oxidase I in the subthalamic nucleus, as expected. In addition, D(2) receptor-deficient mice display a reduced expression of glutamate decarboxylase-67 messenger RNA in the globus pallidus, as the basal ganglia model predicts. This reduction contrasts with the lack of change or increase in glutamate decarboxylase-67 messenger RNA expression found in animals depleted of dopamine after lesions of the mesostriatal dopaminergic system. Furthermore, D(2) receptor-deficient mice show a significant decrease in substance P messenger RNA expression in the striatonigral neurons which form the direct pathway. Finally, glutamate decarboxylase-67 messenger RNA expression in the basal ganglia output nuclei was not affected by mutations in the D(2) receptor gene, a fact that could probably be related to the absence of a parkinsonian locomotor phenotype in D(2) receptor-deficient mice. In summary, these findings provide compelling evidence demonstrating that the lack of endogenous stimulation of D(2) receptors is sufficient to produce subthalamic nucleus hyperactivity, as assessed by cytochrome oxidase I histochemistry and messenger RNA expression, and strongly suggest the existence of interactions between the basal ganglia direct and indirect pathways.
Asunto(s)
Globo Pálido/citología , Neostriado/citología , Receptores de Dopamina D2/genética , Sustancia Negra/citología , Núcleo Subtalámico/citología , Animales , Complejo IV de Transporte de Electrones/análisis , Complejo IV de Transporte de Electrones/genética , Femenino , Expresión Génica/fisiología , Globo Pálido/química , Glutamato Descarboxilasa/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neostriado/química , Vías Nerviosas , Neuronas/química , Neuronas/enzimología , Enfermedad de Parkinson/patología , Fenotipo , ARN Mensajero/análisis , Sustancia P/genética , Sustancia Negra/química , Núcleo Subtalámico/químicaRESUMEN
This study compared the effects of the disruption of the two main presumably glutamatergic striatal inputs, the corticostriatal and thalamostriatal pathways, on GLT1 expression in the rat striatum, using in situ hybridization and immunohistochemistry. Unilateral ibotenate-induced thalamic lesion produced no significant changes in striatal GLT1 mRNA labeling and immunostaining as assessed at 5 and 12 days postlesion. In contrast, significant increases in both parameters were measured after bilateral cortical lesion by superficial thermocoagulation. GLT1 mRNA levels increased predominantly in the dorsolateral part of the striatum; there, the increases were significant at 5 (+84%), 12 (+101%), and 21 (+45%) but not at 35 days postlesion. GLT1 immunostaining increased significantly and homogeneously by 17-26% at 12 and 21 days postlesion. The increase in GLT1 expression at 12 days postlesion was further confirmed by western blot analysis; in contrast, a 36% decrease in glutamate uptake activity was measured at the same time point. These data indicate that striatal GLT1 expression depends on corticostriatal but not thalamostriatal innervation. Comparison of our results with previous data showing that cortical lesion by aspiration downregulates striatal GLT1 expression further suggests that differential changes in GLT1 expression, and thus presumably in glial cell function, may occur in the target striatum depending on the way the cortical neurons degenerate.
Asunto(s)
Corteza Cerebral/fisiología , Cuerpo Estriado/fisiología , Proteínas de Transporte de Monosacáridos/metabolismo , Tálamo/fisiología , Vías Aferentes/fisiología , Animales , Autorradiografía , Cuerpo Estriado/metabolismo , Desnervación , Femenino , Transportador de Glucosa de Tipo 1 , Inmunohistoquímica , Hibridación in Situ , Proteínas de Transporte de Monosacáridos/genética , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Distribución TisularRESUMEN
This study examined the effects of unilateral thermocoagulatory cortical lesion on the pattern of neuropeptide Y immunostaining in the rat ipsilateral striatum at 4 and 21 days post-lesion. Light microscopic analysis showed a significant increase in the number of neuropeptide Y-positive neurons vs. control at both time points; paradoxically, the intraneuronal level of labelling significantly decreased at 4 days post-lesion but increased at 21 days post-lesion. Ultrastructural analysis in control condition showed a higher proportion of dendritic versus axonal labelled processes (3.5 ratio); all the neuropeptide Y synaptic terminals formed symmetrical contacts, mostly onto unlabelled dendrites. At 4 days post-lesion, the neuropeptide Y-positive axon density dramatically increased (+576%) without significant change in the labelled dendrite density, vs. control values; the density of neuropeptide Y synaptic terminals increased in parallel by 233%. In addition, a significant proportion of large neuropeptide Y boutons forming asymmetrical synapses onto unlabelled spines were observed. At 21 days post-lesion, densities of neuropeptide Y dendrites, axons, and synaptic terminals increased by 68, 246 and 125%, respectively, vs. control. But, the morphological features of the neuropeptide Y axonal processes and synaptic specializations of the boutons were similar to those observed in control condition. These data (1) raise an important issue regarding the origin of the terminals forming asymmetrical synapses in the striatum, (2) suggest that adaptative changes in the neuropeptide Y neuronal network may be a main component of striatal remodelling resulting from the progressive loss of cortical inputs, and (3) reinforce the view that neuropeptide Y and excitatory amino acid functions may be tightly linked in the striatum.
Asunto(s)
Neostriado/metabolismo , Neuronas/metabolismo , Neuropéptido Y/análisis , Neuropéptido Y/metabolismo , Animales , Femenino , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Microscopía Electrónica , Neostriado/fisiología , Neostriado/ultraestructura , Neuronas/fisiología , Neuronas/ultraestructura , Ratas , Ratas Wistar , Sinapsis/química , Sinapsis/ultraestructura , Factores de TiempoRESUMEN
Striatin, an intraneuronal, calmodulin-binding protein addressed to dendrites and spines, is expressed in the motor system, particularly the striatum and motoneurons. Striatin contains a high number of domains mediating protein-protein interactions, suggesting a role within a dendritic Ca(2+)-signaling pathway. Here, we explored the hypothesis of a direct role of striatin in the motor control of behaving rats, by using an antisense strategy based on oligodeoxynucleotides (ODN). Rats were treated by intracerebroventricular infusion of a striatin antisense ODN (A-ODN) or mismatch ODN (M-ODN) delivered by osmotic pumps over 6 days. A significant decrease in the nocturnal locomotor activity of A-ODN-treated rats was observed after 5 days of treatment. Hypomotricity was correlated with a 60% decrease in striatin content of the striata of A-ODN-treated rats sacrificed on day 6. Striatin thus plays a role in the control of motor function. To approach the cellular mechanisms in which striatin is involved, striatin down-regulation was studied in a comparatively simpler model: purified rat spinal motoneurons which retain their polarity in culture. Treatment of cells by the striatin A-ODN resulted in the impairement of the growth of dendrites but not axon. The decrease in dendritic growth paralleled the loss of striatin. This model allows analysis of the molecular basis of striatin function in the dynamic changes occurring in growing dendrites, and offers clues to unravel its function within spines.
Asunto(s)
Proteínas de Unión a Calmodulina/fisiología , Cuerpo Estriado/fisiopatología , Regulación de la Expresión Génica/efectos de los fármacos , Locomoción/fisiología , Proteínas de la Membrana/fisiología , Trastornos del Movimiento/fisiopatología , Proteínas del Tejido Nervioso/fisiología , Animales , Calmodulina/metabolismo , Proteínas de Unión a Calmodulina/biosíntesis , Proteínas de Unión a Calmodulina/deficiencia , Proteínas de Unión a Calmodulina/genética , Células Cultivadas , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Dendritas/ultraestructura , Depresión Química , Inyecciones Intraventriculares , Masculino , Proteínas de la Membrana/biosíntesis , Proteínas de la Membrana/deficiencia , Proteínas de la Membrana/genética , Neuronas Motoras/efectos de los fármacos , Neuronas Motoras/patología , Trastornos del Movimiento/genética , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/deficiencia , Proteínas del Tejido Nervioso/genética , Oligonucleótidos Antisentido/administración & dosificación , Oligonucleótidos Antisentido/farmacología , Oligonucleótidos Antisentido/toxicidad , Ratas , Ratas Wistar , Médula Espinal/citologíaRESUMEN
The present study examined whether cortical damage in rats may disrupt the integrative processes and motor control involved in the performance of a reaction time (RT) task. To investigate the nature of the deficits in the conditioned task, rats were subjected, after learning, to a coagulation of pia brain surface of varying extent, including the frontal and parietal cortical areas. They were then tested daily for over one month. The behavioural task required the rats to hold a lever down during a variable and random delay and react quickly to the onset of a visual cue by releasing the lever within a RT limit for food reinforcement. Extensive bilateral cortical lesions had no effect on spontaneous motor activity, but severely impaired RT performance. Latencies to release the lever after the cue were dramatically increased during the first postoperative sessions and gradually returned to baseline levels within 3 weeks, whereas less dramatic but long-lasting increase in premature responding (anticipatory response before the visual cue) was observed throughout the testing sessions. More restricted lesions to the frontoparietal cortex produced a similar pattern of incorrect responding with a faster recovery of delayed responses and a strong deficit in premature responding. The major effects of lesions confined to the rostral pole of the frontal cortex were observed on premature responding, however. The present results demonstrate that the impairment in movement initiation is rapidly recovered within 2-3 weeks even after extensive thermocoagulatory lesions of the frontal and parietal areas. This recovery suggests the involvement of adaptive processes developing progressively and probably reflecting the remarkable synaptic plasticity of the extrapyramidal motor output. In contrast, the long-lasting increase in premature responding, supposed to reflect some attentional deficits, may produce anatomofunctional long-term disorganization of subcortical structures such as the basal ganglia. Interestingly enough, these results show that the rat neocortex supports functions very similar to those of primates and provide a good model for studying these higher functions in operant motor procedures that require prior associative learning and appropriate motor coordination.
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Condicionamiento Psicológico/fisiología , Lóbulo Frontal/fisiología , Lóbulo Parietal/fisiología , Animales , Electrocoagulación , Lóbulo Frontal/patología , Masculino , Actividad Motora/fisiología , Lóbulo Parietal/patología , Ratas , Ratas Wistar , Tiempo de Reacción/fisiologíaRESUMEN
Striatin, a 110-kDa protein, is the first member of the tryptophane-aspartate repeat protein family known to bind calmodulin in the presence of Ca2+. We examined the distribution of striatin and its mRNA in the rat central nervous system (CNS) by using immunocytochemistry and in situ hybridization, respectively. Striatin immunostaining and mRNA labeling patterns are generally concordant. Regions showing the most intense staining are the dorsal striatum, nucleus accumbens (anterior and shell parts), olfactory tubercle, red nucleus, subthalamic nucleus, cranial nerve motor nuclei, and layer IX of the spinal cord (motoneurons). Low levels of both striatin and its mRNA are detected in the cerebral cortex, thalamus, septum, amygdala, hippocampus, midbrain and cerebellum. Striatin-immunoreactive neuronal processes are found predominantly in the structures containing striatin-positive neurons, suggesting that these labeled processes represent dendritic arborization rather than axonal processes. Except for the medial forebrain bundle, all axonal fiber tracts examined are devoid of striatin immunolabeling. These data show that the somatodendritic localization of striatin, previously described in the striatum, may be a main feature of the subcellular distribution of this protein throughout the CNS. Although widely distributed in neurons throughout the rat CNS, striatin is expressed prominently in the structures belonging to the motor system, suggesting that this protein may play a preponderant role in motor control.
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Química Encefálica/fisiología , Proteínas de Unión a Calmodulina/análisis , Proteínas de la Membrana/análisis , Proteínas del Tejido Nervioso/análisis , Animales , Diencéfalo/química , Inmunohistoquímica , Hibridación in Situ , Mesencéfalo/química , ARN Mensajero/análisis , Ratas , Ratas Wistar , Rombencéfalo/química , Sensibilidad y Especificidad , Telencéfalo/químicaRESUMEN
This study examined the effects of thalamo-striatal deafferentation on preprotachykinin and preproenkephalin mRNA expression in the rat neostriatum, using quantitative in situ hybridization histochemistry. Unilateral ibotenate-induced intralaminar thalamic lesion produced a significant decrease in preproenkephalin mRNA levels (-27%) restricted to the ipsilateral striatum at 5 days post-lesion. At 12 days post-lesion, significant decreases in striatal preproenkephalin mRNA expression were found on both brain sides. This post-lesional response was more pronounced in the ipsilateral (-32%) than contralateral (-18%) striatum. All these changes were homogeneously distributed between the dorsolateral and ventromedial parts of the striatum. In parallel, no significant change in preprotachykinin mRNA expression was found at either 5 or 12 days after thalamic lesion, when considering the striatum as a whole. However, at 5 days post-lesion, the regional analysis revealed a slight decrease (-17%) in preprotachykinin mRNA expression, confined to the dorsolateral part of the ipsilateral striatum. These results show that thalamic lesion preferentially affects preproenkephalin vs. preprotachykinin gene expression in the striatum, suggesting, at the first site, a predominant influence of thalamo-striatal inputs on the enkephalin-containing striato-pallidal pathway. However, given that the thalamo-striatal projection is strictly ipsilateral, the bilateralization of the down-regulation of preproenkephalin mRNA expression at 12 days post-lesion suggests an involvement of interhemispheric adaptive mechanisms via cortical networks.