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1.
Nutr Neurosci ; 27(10): 1199-1210, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38294500

RESUMO

OBJECTIVES: Neurons and glial cells are the main functional and structural elements of the brain, and the former depends on the latter for their nutritional, functional and structural organization, as well as for their energy maintenance. METHODS: Glucose is the main metabolic source that fulfills energetic demands, either by direct anaplerosis or through its conversion to metabolic intermediates. Development of some neurodegenerative diseases have been related with modifications in the expression and/or function of glial glucose transporters, which might cause physiological and/or pathological disturbances of brain metabolism. In the present contribution, we summarized the experimental findings that describe the exquisite adjustment in expression and function of glial glucose transporters from physiologic to pathologic metabolism, and its relevance to neurodegenerative diseases. RESULTS: A exhaustive literature review was done in order to gain insight into the role of brain energetics in neurodegenerative disease. This study made evident a critical involvement of glucose transporters and thus brain energetics in the development of neurodegenerative diseases. DISCUSSION: An exquisite adjustment in the expression and function of glial glucose transporters from physiologic to pathologic metabolism is a biochemical signature of neurodegenerative diseases.


Assuntos
Encéfalo , Metabolismo Energético , Proteínas Facilitadoras de Transporte de Glucose , Glucose , Doenças Neurodegenerativas , Neuroglia , Humanos , Doenças Neurodegenerativas/metabolismo , Encéfalo/metabolismo , Glucose/metabolismo , Animais , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Neuroglia/metabolismo , Doenças Metabólicas/metabolismo , Transporte Biológico , Neurônios/metabolismo
2.
Proc Natl Acad Sci U S A ; 118(27)2021 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-34215692

RESUMO

Plant diseases are among the major causes of crop yield losses around the world. To confer disease resistance, conventional breeding relies on the deployment of single resistance (R) genes. However, this strategy has been easily overcome by constantly evolving pathogens. Disabling susceptibility (S) genes is a promising alternative to R genes in breeding programs, as it usually offers durable and broad-spectrum disease resistance. In Arabidopsis, the S gene DMR6 (AtDMR6) encodes an enzyme identified as a susceptibility factor to bacterial and oomycete pathogens. Here, we present a model-to-crop translational work in which we characterize two AtDMR6 orthologs in tomato, SlDMR6-1 and SlDMR6-2. We show that SlDMR6-1, but not SlDMR6-2, is up-regulated by pathogen infection. In agreement, Sldmr6-1 mutants display enhanced resistance against different classes of pathogens, such as bacteria, oomycete, and fungi. Notably, disease resistance correlates with increased salicylic acid (SA) levels and transcriptional activation of immune responses. Furthermore, we demonstrate that SlDMR6-1 and SlDMR6-2 display SA-5 hydroxylase activity, thus contributing to the elucidation of the enzymatic function of DMR6. We then propose that SlDMR6 duplication in tomato resulted in subsequent subfunctionalization, in which SlDMR6-2 specialized in balancing SA levels in flowers/fruits, while SlDMR6-1 conserved the ability to fine-tune SA levels during pathogen infection of the plant vegetative tissues. Overall, this work not only corroborates a mechanism underlying SA homeostasis in plants, but also presents a promising strategy for engineering broad-spectrum and durable disease resistance in crops.


Assuntos
Resistência à Doença/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Homologia de Sequência de Aminoácidos , Solanum lycopersicum/imunologia , Proteínas de Arabidopsis/metabolismo , Biocatálise , Regulação da Expressão Gênica de Plantas , Gentisatos/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/crescimento & desenvolvimento , Mutação/genética , Filogenia , Imunidade Vegetal/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Ácido Salicílico/metabolismo , Transcriptoma/genética , Regulação para Cima , Xanthomonas/fisiologia
3.
J Neurochem ; 165(4): 457-466, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36920226

RESUMO

Glutamate is the major excitatory neurotransmitter in the vertebrate brain, it is critically involved in the function and dysfunction of the central nervous system. The molecular cloning of its ionotropic receptors in the last decade of the past century increased exponentially the interest in this neurotransmitter system. Since then, a plethora of knowledge of the structure, function, and regulation of its receptors and transporters has advanced our understanding of glutamate-mediated neurochemical transactions. Moreover, the characterization of glial glutamate receptors together with the compulsory participation of surrounding astrocytes in glutamate turnover and in the known metabolic coupling with neurons has supported what is now known as the tripartite synapses. The molecular characterization of the various glutamate transporters has also been fundamental for the involvement of glial cells in glutamatergic synapses. Using radial glial cultures, over the years, we have demonstrated an alternative glutamate-mediated signaling system triggered by sodium-dependent glutamate transporters. A detailed account of these findings and the signaling through other glutamate transporters are presented here. The role of this signaling system in the context of glutamatergic transmission is discussed as well as the future directions in the field.


Assuntos
Neuroglia , Neurônios , Neuroglia/metabolismo , Neurônios/metabolismo , Ácido Glutâmico/metabolismo , Sistema X-AG de Transporte de Aminoácidos/metabolismo , Neurotransmissores/metabolismo
4.
J Neurochem ; 163(6): 478-499, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36153691

RESUMO

The developmental process of central nervous system (CNS) myelin sheath formation is characterized by well-coordinated cellular activities ultimately ensuring rapid and synchronized neural communication. During this process, myelinating CNS cells, namely oligodendrocytes (OLGs), undergo distinct steps of differentiation, whereby the progression of earlier maturation stages of OLGs represents a critical step toward the timely establishment of myelinated axonal circuits. Given the complexity of functional integration, it is not surprising that OLG maturation is controlled by a yet fully to be defined set of both negative and positive modulators. In this context, we provide here first evidence for a role of lysophosphatidic acid (LPA) signaling via the G protein-coupled receptor LPA6 as a negative modulatory regulator of myelination-associated gene expression in OLGs. More specifically, the cell surface accessibility of LPA6 was found to be restricted to the earlier maturation stages of differentiating OLGs, and OLG maturation was found to occur precociously in Lpar6 knockout mice. To further substantiate these findings, a novel small molecule ligand with selectivity for preferentially LPA6 and LPA6 agonist characteristics was functionally characterized in vitro in primary cultures of rat OLGs and in vivo in the developing zebrafish. Utilizing this approach, a negative modulatory role of LPA6 signaling in OLG maturation could be corroborated. During development, such a functional role of LPA6 signaling likely serves to ensure timely coordination of circuit formation and myelination. Under pathological conditions as seen in the major human demyelinating disease multiple sclerosis (MS), however, persistent LPA6 expression and signaling in OLGs can be seen as an inhibitor of myelin repair. Thus, it is of interest that LPA6 protein levels appear elevated in MS brain samples, thereby suggesting that LPA6 signaling may represent a potential new druggable pathway suitable to promote myelin repair in MS.


Assuntos
Oligodendroglia , Peixe-Zebra , Camundongos , Animais , Ratos , Humanos , Oligodendroglia/metabolismo , Bainha de Mielina/metabolismo , Neurogênese/fisiologia , Diferenciação Celular/fisiologia , Receptores de Ácidos Lisofosfatídicos
5.
J Neurochem ; 155(1): 7-9, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32681746

RESUMO

Hippocampal excitatory glutamatergic transmission is critically involved in cognitive functions such as learning and memory. A severe impairment of spatial memory is associated with the Alzheimer's disease characteristic augmentation of soluble Amyloid-beta1-42 which in turn leads to glutamatergic neurotransmission dysfunction. As the molecular basis of such correlations has not been completely understood, this Editorial highlights a study in the current issue of the Journal of Neurochemistry in which Yeung and coworkers provide an elegant anatomical study that sheds light into this problematic. Through a rigorous immunohistochemical approach, a sub-regional expression pattern of ionotropic glutamate receptors and vesicular transporters was determined in control and beta amyloid-injected mouse hippocampus. The selected areas participate in information processing and thus, in memory formation. Furthermore, the authors discuss their findings in the context of cognitive deficits present in Alzheimer's disease patients delivering an intuitive analysis of plausible molecular events that disturb proper glutamate signaling. This study takes an important step toward a better understanding of the complexity of Amyloid-beta1-42 and glutamatergic neurotransmission interactions.


Assuntos
Peptídeos beta-Amiloides/farmacologia , Glutamatos/fisiologia , Hipocampo/efeitos dos fármacos , Animais , Humanos , Camundongos , Fragmentos de Peptídeos/farmacologia , Receptores de Glutamato/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Proteína Vesicular 1 de Transporte de Glutamato/metabolismo
6.
J Neurochem ; 155(6): 599-601, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33075150

RESUMO

Neurodegenerative diseases are a worldwide health problem and are a major cause of death and disability. A progressive loss of defined neuronal populations is triggered by a diverse array of stimuli that converge in deficient neurotrophic signaling. Therefore, much effort has been placed in recent years in the characterization of the molecular mechanisms associated with the structure and function of neurotrophins, its receptors, signaling strategies, and their target genes. This Editorial highlights an impressive study by the group of Prof. Ashis K. Mukherjee, a renowned specialist in snake venoms, in which a component of the Indian Cobra N.naja venom with no significant similarity to nerve growth factor, is shown to induce sustained neuritogenesis. An elegant transcriptomic and functional analysis of this component, named Nn-α-elapitoxin, mapped novel domains in mammalian neurotrophic receptors that trigger both conventional and novel signal cascades that support neurite extension in the PC-12 neuronal model system. The authors discuss their findings in the context of the paradoxical neurite outgrowth properties of this toxin which originate in their unique receptor binding site. This study takes an important step towards a better understanding of the complexity of neuronal development and maintenance of the nervous system and provides a potential target to improve neurotrophic signaling, independent of endogenous growth factors, in the diseased brain.


Assuntos
Venenos Elapídicos/metabolismo , Doenças Neurodegenerativas/metabolismo , Proteoma/metabolismo , Receptor trkA/metabolismo , Transcriptoma/fisiologia , Animais , Venenos Elapídicos/genética , Naja , Doenças Neurodegenerativas/genética , Células PC12 , Ligação Proteica/fisiologia , Proteoma/genética , Ratos , Receptor trkA/genética , Venenos de Serpentes
7.
Neurochem Res ; 45(6): 1365-1374, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31363896

RESUMO

Glutamate is the major excitatory amino acid neurotransmitter in the vertebrate brain. It exerts its actions through the activation of specific plasma membrane receptors expressed in neurons and glial cells. Overactivation of glutamate receptors results in neuronal death, known as excitotoxicity. A family of sodium-dependent glutamate transporters enriched in glial cells are responsible of the vast majority of the removal of this amino acid form the synaptic cleft. Therefore, a precise and exquisite regulation of these proteins is required not only for a proper glutamatergic transmission but also for the prevention of an excitotoxic insult. Manganese is a trace element essential as a cofactor for several enzymatic systems, although in high concentrations is involved in the disruption of brain glutamate homeostasis. The molecular mechanisms associated to manganese neurotoxicity have been focused on mitochondrial function, although energy depletion severely compromises the glutamate uptake process. In this context, in this contribution we analyze the effect of manganese exposure in glial glutamate transporters function. To this end, we used the well-established model of chick cerebellar Bergmann glia cultures. A time and dose dependent modulation of [3H]-D-aspartate uptake was found. An increase in the transporter catalytic efficiency, most probably linked to a discrete increase in the affinity of the transporter was detected upon manganese exposure. Interestingly, glucose uptake was reduced by this metal. These results favor the notion of a direct effect of manganese on glial cells, this in turn alters their coupling with neurons and might lead to changes in glutamatergic transmission.


Assuntos
Transportador 1 de Aminoácido Excitatório/metabolismo , Manganês/administração & dosagem , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Animais , Ácido Aspártico/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Embrião de Galinha , Relação Dose-Resposta a Droga
8.
J Biochem Mol Toxicol ; 34(8): e22516, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32363662

RESUMO

The liver is the organ responsible for bisphenol A (BPA) metabolism, an environmental chemical agent. Exposure to this toxin is associated with liver abnormalities and dysfunction. An important role played by excitatory amino acid transporters (EAATs) of the slc1 gene family has been reported in liver injuries. To gain insight into a plausible effect of BPA exposure in the liver glutamate/aspartate transport, using the human hepatoblastoma cell line HepG2, we report a BPA-dependent dynamic regulation of SLC1A3 and SLC1A2. Through the use of radioactive [3 H]- d-aspartate uptake experiments and immunochemical approaches, we characterized time and dose-dependent regulation of the protein levels and function of these transporters after acute exposure to BPA. An increase in nuclear Yin Yang 1 was found. These results suggest an important involvement of the EAATs in liver physiology and its disruption after acute BPA exposure.


Assuntos
Ácido Aspártico/metabolismo , Compostos Benzidrílicos/toxicidade , Transportador 1 de Aminoácido Excitatório/metabolismo , Transportador 2 de Aminoácido Excitatório/metabolismo , Fígado/metabolismo , Fenóis/toxicidade , Transporte Biológico Ativo/efeitos dos fármacos , Células Hep G2 , Humanos , Fator de Transcrição YY1/metabolismo
10.
Molecules ; 24(2)2019 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-30641974

RESUMO

Brown seaweeds contain bioactive compounds that show anti-tumorigenic effects. These characteristics have been repeatedly observed in the Lessoniaceae family. Egregia menziesii, a member of this family, is distributed in the North Pacific and its properties have been barely studied. We evaluated herein the cytotoxic and anti-proliferative activity of extracts of this seaweed, through toxicity assay in Artemia salina and lymphocytes, and MTT proliferation assay, in Bergmann glia cells, 3T3-L1 and brain cancer cell lines. E. menziesii's extracts inhibited the spread of all the tested cell lines. The hexane extract showed the highest cytotoxic activity, while the methanol extract was moderately cytotoxic. Interestingly, seaweed extracts displayed a selective inhibition pattern. These results suggest that E. menziesii's extracts might be good candidates for cancer prevention and the development of novel chemotherapies due to its highest cytotoxicity in transformed cells compare to glia primary cultures.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Alga Marinha/química , Animais , Neoplasias Encefálicas , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Embrião de Galinha , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Linfócitos/imunologia , Linfócitos/metabolismo , Camundongos , Ratos
11.
Neural Plast ; 2017: 8140737, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29435373

RESUMO

Circadian rhythms are biological oscillations with a period of about 24 hours. These rhythms are maintained by an innate genetically determined time-keeping system called the circadian clock. A large number of the proteins involved in the regulation of this clock are transcription factors controlling rhythmic transcription of so-called clock-controlled genes, which participate in a plethora of physiological functions in the organism. In the brain, several areas, besides the suprachiasmatic nucleus, harbor functional clocks characterized by a well-defined time pattern of clock gene expression. This expression rhythm is not restricted to neurons but is also present in glia, suggesting that these cells are involved in circadian rhythmicity. However, only certain glial cells fulfill the criteria to be called glial clocks, namely, to display molecular oscillators based on the canonical clock protein PERIOD, which depends on the suprachiasmatic nucleus for their synchronization. In this contribution, we summarize the current information about activity of the clock genes in glial cells, their potential role as oscillators as well as clinical implications.


Assuntos
Encéfalo/citologia , Relógios Circadianos , Ritmo Circadiano , Neuroglia/fisiologia , Animais , Regulação da Expressão Gênica , Humanos , Neurônios/fisiologia
12.
Neurochem Res ; 41(7): 1797-805, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27017513

RESUMO

Glutamate, the main excitatory amino acid in the vertebrate retina, is a well know activator of numerous signal transduction pathways, and has been critically involved in long-term synaptic changes acting through ionotropic and metabotropic glutamate receptors. However, recent findings underlining the importance of intensity and duration of glutamate stimuli for specific neuronal responses, including excitotoxicity, suggest a crucial role for Na(+)-dependent glutamate transporters, responsible for the removal of this neurotransmitter from the synaptic cleft, in the regulation of glutamate-induced signaling. Transporter proteins are expressed in neurons and glia cells, albeit most of glutamate uptake occurs in the glial compartment. Within the retina, Müller glia cells are in close proximity to glutamatergic synapses and participate in the recycling of glutamate through the glutamate/glutamine shuttle. In this context, we decided to investigate a plausible role of glutamate as a regulatory signal for its own transport in human retinal glia cells. To this end, we determined [(3)H]-D-aspartate uptake in cultures of spontaneously immortalized human Müller cells (MIO-M1) exposed to distinct glutamatergic ligands. A time and dose-dependent increase in the transporter activity was detected. This effect was dependent on the activation of the N-methyl D-aspartate subtype of glutamate receptors, due to a dual effect: an increase in affinity and an augmented expression of the transporter at the plasma membrane, as established via biotinylation experiments. Furthermore, a NMDA-dependent association of glutamate transporters with the cystoskeletal proteins ezrin and glial fibrillary acidic protein was also found. These results add a novel mediator of the glutamate transporter modulation and further strengthen the notion of the critical involvement of glia cells in synaptic function.


Assuntos
Células Ependimogliais/metabolismo , Ácido Glutâmico/metabolismo , Neuroglia/metabolismo , Receptores de Glutamato/metabolismo , Regulação para Cima/fisiologia , Ácido Aspártico/metabolismo , Ácido Aspártico/farmacologia , Células Cultivadas , Células Ependimogliais/efeitos dos fármacos , Agonistas de Aminoácidos Excitatórios/farmacologia , Humanos , Neuroglia/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos
13.
Neurochem Res ; 40(5): 961-70, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25749891

RESUMO

Glutamate, the major excitatory amino acid, activates a wide variety of signal transduction cascades. This neurotransmitter is involved in photic entrainment of circadian rhythms, which regulate physiological and behavioral functions. The circadian clock in vertebrates is based on a transcription-translation feedback loop in which Brain and muscle aryl hydrocarbon receptor nuclear translocator (ARNT)-like protein 1 (BMAL1) acts as transcriptional activator of others clock genes. This protein is expressed in nearly all suprachiasmatic nucleus neurons, as well as in the granular layer of the cerebellum. In this context, we decided to investigate the role of glutamate in the molecular mechanisms involved in the processes of transcription/translation of BMAL1 protein. To this end, primary cultures of chick cerebellar Bergmann glial cells were stimulated with glutamatergic ligands and we found that BMAL1 levels increased in a dose- and time dependent manner. Additionally, we studied the phosphorylation of serine residues in BMAL1 under glutamate stimulation and we were able to detect an increase in the phosphorylation of this protein. The increased expression of BMAL1 is most probably the result of a stabilization of the protein after it has been phosphorylated by the cyclic AMP-dependent protein kinase and/or the Ca(2+)/diacylglycerol dependent protein kinase. The present results strongly suggest that glutamate participates in regulating BMAL1 in glial cells and that these cells might prove to be important in the control of circadian rhythms in the cerebellum.


Assuntos
Fatores de Transcrição ARNTL/fisiologia , Ácido Glutâmico/farmacologia , Neuroglia/efeitos dos fármacos , Neuroglia/fisiologia , Animais , Células Cultivadas , Embrião de Galinha , Relação Dose-Resposta a Droga , Transdução de Sinais
14.
Neurochem Res ; 40(11): 2317-24, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26384974

RESUMO

Glutamate, the main excitatory transmitter in the vertebrate brain, exerts its actions through the activation of specific membrane receptors present in neurons and glial cells. Over-stimulation of glutamate receptors results in neuronal death, phenomena known as excitotoxicity. A family of glutamate uptake systems, mainly expressed in glial cells, removes the amino acid from the synaptic cleft preventing an excessive glutamatergic stimulation and thus neuronal damage. Autism spectrum disorders comprise a group of syndromes characterized by impaired social interactions and anxiety. One or the most common drugs prescribed to treat these disorders is Methylphenidate, known to increase dopamine extracellular levels, although it is not clear if its sedative effects are related to a plausible regulation of the glutamatergic tone via the regulation of the glial glutamate uptake systems. To gain insight into this possibility, we used the well-established model system of cultured chick cerebellum Bergmann glia cells. A time and dose-dependent increase in the activity and protein levels of glutamate transporters was detected upon Methylphenidate exposure. Interestingly, this increase is the result of an augmentation of both the synthesis as well as the insertion of these protein complexes in the plasma membrane. These results favour the notion that glial cells are Methylphenidate targets, and that by these means could regulate dopamine turnover.


Assuntos
Inibidores da Captação de Dopamina/farmacologia , Ácido Glutâmico/metabolismo , Metilfenidato/farmacologia , Neuroglia/metabolismo , Animais , Ácido Aspártico/metabolismo , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Embrião de Galinha , Dopamina/metabolismo , Transportador 1 de Aminoácido Excitatório/metabolismo , Neuroglia/efeitos dos fármacos , RNA/biossíntese , RNA/isolamento & purificação , Regulação para Cima/efeitos dos fármacos
15.
Neurochem Res ; 40(5): 915-23, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25736255

RESUMO

Glutamate (Glu) the main excitatory neurotransmitter of the central nervous system regulates gene expression at different levels through the activation of specific membrane receptors and transporters expressed in neurons and glia cells. A membrane to nucleus signaling cascade triggered by this neurotransmitter has been described in cultured cerebellar Bergmann glia cells isolated from chick embryos. Furthermore, it has also been described that Glu receptors activation is linked to a modulation of [(35)S]-methionine incorporation into newly synthesized polypeptides. In order to gain insight into the signal transduction cascades that participate in this effect, in the present study we characterized the phosphorylation of a critical component of the translational machinery, namely the ribosomal protein S6. The phosphorylation sites in rpS6 have been mapped to five clustered residues, Ser235, Ser236, Ser240, Ser244 and Ser247. Nevertheless, Ser236 phosphorylation is the primary phosphorylation site. The kinases responsible of this modification are p70(S6K) and p90(RSK). rpS6 phosphorylation increases the affinity of 40s subunit for mRNAs and thus facilitates translational initiation. Glutamate exposure of cultured cerebellar Bergmann glia cells results in a time- and dose-dependent increase in rpS6 phosphorylation. This effect is mainly observed at cytoplasm, and involves the phosphoinositol-3 kinase/protein kinase B pathway. Our results favor the notion of a continuous neuronal signaling to glia cells that regulates the proteome of these cells not only at the transcriptional level but also at the level of protein synthesis.


Assuntos
Ácido Glutâmico/farmacologia , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos , Biossíntese de Proteínas/fisiologia , Proteína S6 Ribossômica/metabolismo , Animais , Células Cultivadas , Embrião de Galinha , Relação Dose-Resposta a Droga , Fosforilação/efeitos dos fármacos , Fosforilação/fisiologia , Proteína S6 Ribossômica/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia
16.
Neural Plast ; 2015: 787396, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26345375

RESUMO

Glutamatergic transmission in the vertebrate brain requires the involvement of glia cells, in a continuous molecular dialogue. Glial glutamate receptors and transporters are key molecules that sense synaptic activity and by these means modify their physiology in the short and long term. Posttranslational modifications that regulate protein-protein interactions and modulate transmitter removal are triggered in glial cells by neuronal released glutamate. Moreover, glutamate signaling cascades in these cells are linked to transcriptional and translational control and are critically involved in the control of the so-called glutamate/glutamine shuttle and by these means in glutamatergic neurotransmission. In this contribution, we summarize our current understanding of the biochemical consequences of glutamate synaptic activity in their surrounding partners and dissect the molecular mechanisms that allow neurons to take control of glia physiology to ensure proper glutamate-mediated neuronal communication.


Assuntos
Glutamatos/fisiologia , Transmissão Sináptica/fisiologia , Animais , Encéfalo/fisiologia , Humanos , Doenças do Sistema Nervoso/fisiopatologia , Neuroglia/fisiologia , Receptores de Glutamato/fisiologia , Proteínas Vesiculares de Transporte de Glutamato/metabolismo , Proteínas Vesiculares de Transporte de Glutamato/fisiologia
17.
Biochim Biophys Acta ; 1833(3): 698-711, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23220011

RESUMO

We recently characterized a nuclear import pathway for ß-dystroglycan; however, its nuclear role remains unknown. In this study, we demonstrate for the first time, the interaction of ß-dystroglycan with distinct proteins from different nuclear compartments, including the nuclear envelope (NE) (emerin and lamins A/C and B1), splicing speckles (SC35), Cajal bodies (p80-coilin), and nucleoli (Nopp140). Electron microscopy analysis revealed that ß-dystroglycan localized in the inner nuclear membrane, nucleoplasm, and nucleoli. Interestingly, downregulation of ß-dystroglycan resulted in both mislocalization and decreased expression of emerin and lamin B1, but not lamin A/C, as well in disorganization of nucleoli, Cajal bodies, and splicing speckles with the concomitant decrease in the levels of Nopp140, and p80-coilin, but not SC35. Quantitative reverse transcription PCR and cycloheximide-mediated protein arrest assays revealed that ß-dystroglycan deficiency did not change mRNA expression of NE proteins emerin and lamin B1 bud did alter their stability, accelerating protein turnover. Furthermore, knockdown of ß-dystroglycan disrupted NE-mediated processes including nuclear morphology and centrosome-nucleus linkage, which provides evidence that ß-dystroglycan association with NE proteins is biologically relevant. Unexpectedly, ß-dystroglycan-depleted cells exhibited multiple centrosomes, a characteristic of cancerous cells. Overall, these findings imply that ß-dystroglycan is a nuclear scaffolding protein involved in nuclear organization and NE structure and function, and that might be a contributor to the biogenesis of nuclear envelopathies.


Assuntos
Nucléolo Celular/metabolismo , Núcleo Celular/ultraestrutura , Corpos Enovelados/metabolismo , Distroglicanas/metabolismo , Mioblastos/metabolismo , Membrana Nuclear/metabolismo , Animais , Western Blotting , Nucléolo Celular/genética , Núcleo Celular/metabolismo , Células Cultivadas , Corpos Enovelados/genética , Distroglicanas/genética , Imunofluorescência , Imunoprecipitação , Lamina Tipo B/genética , Lamina Tipo B/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Músculo Esquelético/citologia , Músculo Esquelético/metabolismo , Músculo Esquelético/ultraestrutura , Mioblastos/citologia , Mioblastos/ultraestrutura , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
18.
Glia ; 62(9): 1543-1558, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24866099

RESUMO

Signaling via the major excitatory amino acid glutamate has been implicated in the regulation of various aspects of the biology of oligodendrocytes, the myelinating cells of the central nervous system (CNS). In this respect, cells of the oligodendrocyte lineage have been described to express a variety of glutamate-responsive transmembrane proteins including sodium-dependent glutamate transporters. The latter have been well characterized to mediate glutamate clearance from the extracellular space. However, there is increasing evidence that they also mediate glutamate-induced intracellular signaling events. Our data presented here show that the activation of oligodendrocyte expressed sodium-dependent glutamate transporters, in particular GLT-1 and GLAST, promotes the morphological aspects of oligodendrocyte maturation. This effect was found to be associated with a transient increase in intracellular calcium levels and a transient phosphorylation event at the serine (S)(371) site of the calcium sensor calcium/calmodulin-dependent kinase type IIß (CaMKIIß). The potential regulatory S(371) site is located within CaMKIIß's previously defined actin-binding/-stabilizing domain, and phosphorylation events within this domain were identified in our studies as a requirement for sodium-dependent glutamate transporter-mediated promotion of oligodendrocyte maturation. Furthermore, our data provide good evidence for a role of these phosphorylation events in mediating detachment of CaMKIIß from filamentous (F)-actin, and hence allowing a remodeling of the oligodendrocyte's actin cytoskeleton. Taken together with our recent findings, which demonstrated a crucial role of CaMKIIß in regulating CNS myelination in vivo, our data strongly suggest that a sodium-dependent glutamate transporter-CaMKIIß-actin cytoskeleton axis plays an important role in the regulation of oligodendrocyte maturation and CNS myelination.


Assuntos
Actinas/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Transportador 1 de Aminoácido Excitatório/metabolismo , Transportador 2 de Aminoácido Excitatório/metabolismo , Oligodendroglia/citologia , Oligodendroglia/fisiologia , Citoesqueleto de Actina/fisiologia , Animais , Encéfalo/citologia , Encéfalo/fisiologia , Cálcio/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/genética , Células Cultivadas , Ácido Glutâmico/metabolismo , Espaço Intracelular/metabolismo , Fosforilação , Ratos Sprague-Dawley , Sódio/metabolismo
19.
Neurochem Res ; 39(1): 142-9, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24248861

RESUMO

Glutamate, the major excitatory neurotransmitter in the vertebrate brain, is a potent neurotoxin therefore its extracellular levels have to be tightly regulated by means of sodium-dependent glutamate uptake systems of the slc1A family. The glial glutamate/aspartate transporter (GLAST/EAAT1) and the glutamate transporter 1 carry most of the uptake activity in cerebellum and in the forebrain, respectively. In the cerebellar cortex, GLAST is profusely expressed in Bergmann glia cells, which completely enwrap the parallel fiber-Purkinje cells synapses. Glutamate exposure in these cells, down regulates the activity as well as the expression levels of this transporter. In order to characterize the persistence of a single glutamate exposure, we followed the [(3)H]-D-aspartate uptake activity as a function of time after the removal of the glutamatergic stimulus. We were able to demonstrate that a single 30 min exposure to glutamate reduces the uptake activity for up to 3 h. This effect is dose-dependent and it is not reproduced neither by ionotropic nor metabotropic glutamate receptors agonists. In contrast, transporter specific ligands such as D-aspartate or L-(-)-threo-3-Hydroxyaspartic acid fully reproduce the glutamate effect. Equilibrium binding experiments revealed a decrease in [(3)H]-D-aspartate Bmax without a significant change in affinity, clearly suggesting that a reduction in the availability of plasma membrane glutamate transporters is the molecular basis of this effect. Interestingly, neither Glast mRNA nor its protein levels were significantly reduced upon the single glutamate exposure. Taken together, these results favor the notion of a transporter-mediated tight control of the uptake process.


Assuntos
Transportador 1 de Aminoácido Excitatório/metabolismo , Ácido Glutâmico/farmacologia , Animais , Ácido Aspártico/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Cerebelo/metabolismo , Embrião de Galinha , Regulação para Baixo , Transportador 1 de Aminoácido Excitatório/efeitos dos fármacos , Neuroglia/metabolismo
20.
Mem Inst Oswaldo Cruz ; 109(2): 174-81, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24676665

RESUMO

Chagas disease, caused by Trypanosoma cruzi, represents an endemic among Latin America countries. The participation of free radicals, especially nitric oxide (NO), has been demonstrated in the pathophysiology of seropositive individuals with T. cruzi. In Chagas disease, increased NO contributes to the development of cardiomyopathy and megacolon. Metallothioneins (MTs) are efficient free radicals scavengers of NO in vitro and in vivo. Here, we developed a murine model of the chronic phase of Chagas disease using endemic T. cruzi RyCH1 in BALB/c mice, which were divided into four groups: infected non-treated (Inf), infected N-monomethyl-L-arginine treated (Inf L-NAME), non-infected L-NAME treated and non-infected vehicle-treated. We determined blood parasitaemia and NO levels, the extent of parasite nests in tissues and liver MT-I expression levels. It was observed that NO levels were increasing in Inf mice in a time-dependent manner. Inf L-NAME mice had fewer T. cruzi nests in cardiac and skeletal muscle with decreased blood NO levels at day 135 post infection. This affect was negatively correlated with an increase of MT-I expression (r = -0.8462, p < 0.0001). In conclusion, we determined that in Chagas disease, an unknown inhibitory mechanism reduces MT-I expression, allowing augmented NO levels.


Assuntos
Doença de Chagas/sangue , Metalotioneína/sangue , Óxido Nítrico/sangue , Animais , Antioxidantes/análise , Doença de Chagas/tratamento farmacológico , Modelos Animais de Doenças , Inibidores Enzimáticos/uso terapêutico , Feminino , Coração/parasitologia , Camundongos Endogâmicos BALB C , Músculo Esquelético/patologia , Miocárdio/patologia , NG-Nitroarginina Metil Éster/uso terapêutico , Estresse Oxidativo , Parasitemia/sangue , Parasitemia/fisiopatologia , Reação em Cadeia da Polimerase em Tempo Real , Estatísticas não Paramétricas , Fatores de Tempo , Trypanosoma cruzi
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