RESUMO
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.
Assuntos
Córtex Cerebral/fisiologia , Corpo Estriado/fisiologia , Proteínas de Transporte de Monossacarídeos/metabolismo , Tálamo/fisiologia , Vias Aferentes/fisiologia , Animais , Autorradiografia , Corpo Estriado/metabolismo , Denervação , Feminino , Transportador de Glucose Tipo 1 , Imuno-Histoquímica , Hibridização In Situ , Proteínas de Transporte de Monossacarídeos/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
Previous studies have shown that the striatum expresses very low levels of Na+/Cl(-)-dependent "orphan" transporter Rxt1 transcripts but contains high levels of protein. This study investigated the origin of Rxt1 expression in rat striatum. Striatal Rxt1 contents assessed by immunocytochemistry or western blotting were found to be significantly reduced after corticostriatal denervation but not after striatal or thalamic lesion with kainic acid or selective 6-hydroxydopamine-induced nigrostriatal deafferentation. Corticostriatal neurons retrogradely labeled by intrastriatal fluorogold injections were shown to express Rxt1 mRNA. Combination of anterograde biotin-dextran amine labeling of the corticostriatal pathway with Rxt1 immunogold detection at the ultrastructural level demonstrated the presence of Rxt1 in about one-third of the corticostriatal synaptic terminals and in numerous unidentified synaptic terminals. All the Rxt1-positive terminals formed asymmetrical contacts on spines. These data provide evidence that striatal Rxt1 immunoreactivity is mainly of extrinsic origin and more specifically associated with the corticostriatal pathway. Rxt1 appears as a selective presynaptic marker of synapses formed by presumably excitatory amino acid afferents, but it segregates a subclass of these synapses, thereby revealing a functional heterogeneity among excitatory amino acid systems.
Assuntos
Proteínas de Transporte/genética , Córtex Cerebral/citologia , Corpo Estriado/citologia , Proteínas de Membrana Transportadoras , Terminações Nervosas/química , Proteínas do Tecido Nervoso/genética , Vias Aferentes , Animais , Autorradiografia , Biotina/análogos & derivados , Química Encefálica , Córtex Cerebral/química , Corpo Estriado/química , Denervação , Dextranos , Agonistas de Aminoácidos Excitatórios , Corantes Fluorescentes , Hibridização In Situ , Ácido Caínico , Masculino , Microscopia Eletrônica , Terminações Nervosas/ultraestrutura , Oxidopamina , Proteínas da Membrana Plasmática de Transporte de Neurotransmissores , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Substância Negra/citologia , Simpatolíticos , Sinapses/química , Tálamo/citologiaRESUMO
Striatin, a recently isolated rat brain calmodulin-binding protein belonging to the WD-repeat protein family, is thought to be part of a calcium signal transduction pathway presumably specific to excitatory synapses, at least in the striatum. This study was aimed to specify the cellular and subcellular localization of striatin, and to determine the possible synaptic relationships between the two main excitatory afferent pathways, arising from the cerebral cortex and the thalamus, and the striatin-containing elements, in the rat striatum. Anterograde tract-tracing by means of biotinylated dextran amine injection in the frontoparietal cerebral cortex or the parafascicular nucleus of the thalamus was combined with immunogold detection of striatin. Striatin-immunoreactivity was confined to the neuronal somatodendritic compartment, including spines. Whereas 90-95% of the striatal neurons were striatin-positive, only about 50% of the sections of dendritic spines engaged in asymmetrical synaptic contacts exhibited striatin labelling. Among the sections of striatin-immunopositive dendritic spines, the number of immunogold particles ranged from one to more than seven, indicating an heterogeneity of the spine labelling. Moreover, within each class of spines presenting at least two silver-gold particles, the distribution of the particles varied from a clear-cut alignment under the postsynaptic densities (24-33% of spines) to a location distant from the synaptic area. In the cell bodies and dendrites, striatin labelling was usually not associated with the cytoplasmic membrane nor with the postsynaptic densities. In the striatum ipsilateral to the tracer injections, only 34.8% of the synaptic contacts formed by corticostriatal afferents involved striatin-positive elements (slightly labelled dendritic spines), whereas 56.7% of the synaptic contacts formed by thalamostriatal boutons were made on striatin-positive targets (mostly dendrites). In both cases, striatin labelling was usually not associated with the postsynaptic density. Most of the immunoreactive dendritic spines were in contact with unidentified afferents. These data reveal that striatin is expressed in the vast majority of the cell bodies of striatal spiny neurons, but is heterogeneously distributed among the dendritic spines of those neurons. Data also indicate a preferential relationship between striatin-containing structures and afferents from the parafascicular thalamic nucleus with respect to the frontoparietal cerebral cortex. But, at the dendritic spine level, striatin may be involved in signal transduction mechanisms involving as yet unidentified excitatory afferents to striatal neurons.
Assuntos
Proteínas de Ligação a Calmodulina/metabolismo , Corpo Estriado/metabolismo , Neurônios/fisiologia , Tálamo/metabolismo , Vias Aferentes/metabolismo , Animais , Corpo Estriado/citologia , Corpo Estriado/ultraestrutura , Feminino , Microscopia Eletrônica , Terminações Nervosas/ultraestrutura , Fibras Nervosas/metabolismo , Neurônios Aferentes/ultraestrutura , Ratos , Ratos Wistar , Tálamo/citologiaRESUMO
This study examined at the ultrastructural level the putative relationships between afferent fibres coming from the parafascicular nucleus of the thalamus and neuropeptide Y (NPY)-containing neurones in the rat striatum. Experiments used a combination of anterograde transport of the biotin dextran amine to label the thalamo-striatal pathway and immunogold labelling to reveal the NPY-containing neurones at the electron microscopic level. Examination of sections from three animals failed to demonstrate thalamic terminals in synaptic contact with NPY-immunoreactive dendrites or cell bodies, although both types of labelled elements were frequently involved in synaptic complex with unlabelled profiles. These results strongly suggest that striatal NPY interneurones are not under the direct influence of the main component of the thalamo-striatal system.
Assuntos
Vias Aferentes/ultraestrutura , Corpo Estriado/ultraestrutura , Neuropeptídeo Y/metabolismo , Tálamo/ultraestrutura , Animais , Feminino , Imuno-Histoquímica , Microscopia Eletrônica , Ratos , Ratos WistarRESUMO
Kinetic parameters of 3H-choline, 3H-GABA and 3H-dopamine (DA) uptakes in striatal homogenates containing nerve endings were determined 2 to 3 weeks after kainic acid injection into the ipsilateral "centre médian"-parafascicular complex area of the thalamus in the rat. Results showed a marked decrease in 3H-choline uptake concomitant with a selective decrease in Vmax. Data also showed a large decrease in 3H-GABA uptake resulting from a decreased affinity of uptake sites for their substrate. These data were associated with the previously described decrease in choline acetyltransferase and increase in glutamic acid decarboxylase apparent activity, respectively. An apparent marked increase in 3H-DA uptake was likewise measured, mainly related to an increase in Vmax. Determination of serotonin (5HT) and 5-hydroxyindole acetic acid (5HIAA) endogenous contents showed in the deafferented striatum a decrease in 5HT concentrations associated with an increase in 5HIAA levels. Taken together, all these changes in neurotransmitter markers suggest that, directly through the thalamostriatal pathway or indirectly, the thalamus can exert a complex influence on striatal cholinergic and GABAergic neuronal functions as well as on the activity of dopaminergic and serotoninergic striatal afferent fibers.