RESUMO
Glutamate is the major excitatory neurotransmitter in the vertebrate central nervous system. During synaptic activity, glutamate is released and binds to specific membrane receptors and transporters activating, in the one hand, a wide variety of signal transduction cascades, while in the other hand, its removal from the synaptic cleft. Extracellular glutamate concentrations are maintained within physiological levels mainly by glia glutamate transporters. Inefficient clearance of this amino acid is neurotoxic due to a prolonged hyperactivation of its postsynaptic receptors, exacerbating a wide array of intracellular events linked to an ionic imbalance, that results in neuronal cell death. This process is known as excitotoxicity and is the underlying mechanisms of an important number of neurodegenerative diseases. Therefore, it is important to understand the regulation of glutamate transporters function. The transporter activity can be regulated at different levels: gene expression, transporter protein targeting and trafficking, and post-translational modifications of the transporter protein. The identification of these mechanisms has paved the way to our current understanding the role of glutamate transporters in brain physiology and will certainly provide the needed biochemical information for the development of therapeutic strategies towards the establishment of novel therapeutic approaches for the treatment and/or prevention of pathologies associated with excitotoxicity insults. This article is part of the issue entitled 'Special Issue on Neurotransmitter Transporters'.
Assuntos
Sistema X-AG de Transporte de Aminoácidos/genética , Sistema X-AG de Transporte de Aminoácidos/fisiologia , Regulação da Expressão Gênica/genética , Regulação da Expressão Gênica/fisiologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Sistema X-AG de Transporte de Aminoácidos/biossíntese , Animais , Glutamatos/fisiologia , Humanos , Neurotransmissores/fisiologiaRESUMO
Glutamate, the main excitatory amino acid in the central nervous system, elicits its functions through the activation of specific membrane receptors that are expressed in neurons and glial cells. The re-cycling of this amino acid is carried out mostly through a continuous interplay between neurons and glia cells, given the fact that the removal of glutamate from the synaptic cleft depends mainly on glial glutamate transporters. Therefore, a functional and physical interaction between membrane transporters links glutamate uptake, transformation to glutamine and its release to the extra-synaptic space and its uptake to the pre-synaptic terminal. This sequence of events, best known as the glutamate/glutamine shuttle is central to glutamatergic transmission. In this sense, the uptake process triggers a complex series of biochemical cascades that modify the physiology of glial cells in the immediate, short and long term so as to be capable to take up, transform and release these amino acids in a regulated amount and in an appropriate time frame to sustain glutamatergic neurotransmission. Among the signaling cascades activated in glial cells by glutamate transporters, a sustained Na(+) and Ca(2+) influx, protein posttranslational modifications and gene expression regulation at the transcriptional and translational levels are present. Therefore, it is clear that the pivotal role of glial cells in the context of excitatory transmission has been constantly underestimated.
Assuntos
Sistema X-AG de Transporte de Aminoácidos/metabolismo , Membrana Celular/metabolismo , Ácido Glutâmico/fisiologia , Proteínas de Membrana Transportadoras/metabolismo , Neuroglia/metabolismo , Transmissão Sináptica/genética , Transmissão Sináptica/fisiologia , Sistema X-AG de Transporte de Aminoácidos/biossíntese , Sistema X-AG de Transporte de Aminoácidos/genética , Animais , Perfilação da Expressão Gênica , Humanos , Proteínas de Membrana Transportadoras/biossíntese , Proteínas de Membrana Transportadoras/genéticaRESUMO
Glutamate, the major excitatory amino acid, activates a wide variety of signal transduction cascades. Ionotropic and metabotropic glutamate receptors are critically involved in long-term synaptic changes, although recent findings suggest that the electrogenic Na(+)-dependent glutamate transporters, responsible for its removal from the synaptic cleft participate in the signaling transactions triggered by this amino acid. Glutamate transporters are profusely expressed in glia therefore most of its uptake occurs in this cellular compartment. In the cerebellar cortex, Bergmann glial cells enwrap glutamatergic synapses and participate in the recycling of its neurotransmitter through the glutamate/glutamine shuttle. It has long been acknowledged that glutamatergic transmission in the cerebellar molecular layer results in cGMP accumulation within Bergmann glia cells. In this context, we decided to investigate a plausible role of the nitric oxide/cGMP-signaling pathway in the regulation of Bergmann glia glutamate transporters. To this end, the well-established model of primary cultures of chick cerebellar Bergmann glial cells was used. Confluent monolayers were exposed to the nitric oxide donor, sodium nitroprusside, or to the non-hydrolysable cGMP analog dbcGMP and the [(3)H] D-aspartate uptake activity measured. An increase in uptake activity, related to an augmentation in VMax, was detected with both treatments. The signaling cascade includes NO/cGMP/PKG and Ca(2+) influx through the Na(+)/Ca(2+) exchanger and might be related to the plasma membrane glutamate transporters turnover. Interestingly enough, an inhibitor of the cGMP dependent protein kinase was capable to abolish the sodium nitroprusside induced Ca(2+) influx. These results provide an insight into the physiological role of cGMP in the cerebellum.
Assuntos
Sistema X-AG de Transporte de Aminoácidos/biossíntese , GMP Cíclico/fisiologia , Neuroglia/metabolismo , Óxido Nítrico/fisiologia , Sistema X-AG de Transporte de Aminoácidos/genética , Animais , Ácido Aspártico/metabolismo , Sinalização do Cálcio/fisiologia , Células Cultivadas , Embrião de Galinha , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , Cultura Primária de Células , Transdução de Sinais/fisiologiaRESUMO
Thyroid hormone (T(3)) regulates the growth and differentiation of rat cerebellar astrocytes. Previously, we have demonstrated that these effects are due, at least in part, to the increased expression of extracellular matrix molecules and growth factors, such as fibroblast growth factor-2. T(3) also modulates neuronal development in an astrocyte-mediated manner. In the mammalian central nervous system, excitatory neurotransmission is mediated mainly by glutamate. However, excessive stimulation of glutamate receptors can lead to excitotoxicity and cell death. Astrocytic glutamate transporters, GLT-1 and GLAST, play an essential role in the clearance of the neuronal-released glutamate from the extracellular space and are essential for maintaining physiological extracellular glutamate levels in the brain. In the present study, we showed that T(3) significantly increased glutamate uptake by cerebellar astrocytes compared with control cultures. Inhibitors of glutamate uptake, such as L-PDC and DL-TBOA, abolished glutamate uptake on control or T(3)-treated astrocytes. T(3) treatment of astrocytes increased both mRNA levels and protein expression of GLAST and GLT-1, although no significant changes on the distribution of these transporters were observed. The gliotoxic effect of glutamate on cultured cerebellar astrocytes was abolished by T(3) treatment of astrocytes. In addition, the neuronal viability against glutamate challenge was enhanced on T(3)-treated astrocytes, showing a putative neuroprotective effect of T(3). In conclusion, our results showed that T(3) regulates extracellular glutamate levels by modulating the astrocytic glutamate transporters. This represents an important mechanism mediated by T(3) on the improvement of astrocytic microenvironment in order to promote neuronal development and neuroprotection.