Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/108666 https://doi.org/10.1186/s12974-016-0613-8 |
Resumo: | Background: Elevated levels of oncostatin M (OSM), an interleukin-6 cytokine family member, have been observed in HIV-1-associated neurocognitive disorders (HAND) and Alzheimer’s disease. However, the function of OSM in these disease conditions is unclear. Since deficient glutamate uptake by astrocytes is instrumental in HAND-associated neurotoxicity, we hypothesized that OSM impairs glutamate uptake in astrocytes and thereby promotes neuronal excitotoxicity. Methods: Primary cultures of mouse cortical astrocytes, neurons, microglia, and BV2 cell line were used. The expression of glutamate transporters (GLAST/EAAT1 and GLT-1/EAAT2) was investigated using real-time PCR and Western blot, and their activity was assessed by measuring 3H-D-aspartate uptake. Neuronal toxicity was measured using the colorimetric MTT (3-(4,5-dimethylthiazol-2-yl-) 2,5-diphenyltetrazolium bromide) assay and immunocytochemistry. A chimeric HIV-1 that infects murine cells (EcoHIV/NL4-3-GFP virus (EcoHIV)) was used to investigate whether the virus induces OSM, OSM receptor (OSMR)-β, glycoprotein 130 (gp130), GLT-1, GLAST (mRNA and protein), and OSM release (ELISA) in cultured BV2 cells, primary microglia, or astrocytes. Statistical analyses of the data were performed using one-way ANOVA (to allow multiple comparisons) and two-tailed Student’s t test. Results: OSM treatment (10 ng/mL) time-dependently reduced GLAST and GLT-1 expression and inhibited 3H-D-aspartate uptake in cultured astrocytes in a concentration-dependent manner, an effect prevented by the Janus kinase (JAK)/signal transducers and activators of transcription (STAT)3 inhibitor AG490. Down-regulation of astrocytic glutamate transport by OSM resulted in NMDA receptor-dependent excitotoxicity in cortical neurons. Infection with EcoHIV induced OSM gene expression and protein release in BV2 cells and microglia, but not in astrocytes. Conversely, EcoHIV caused a fivefold increase in OSMR-β mRNA (but not gp130) and protein in astrocytes, but not in microglia, which did not express OSMR-β protein. Finally, astrocytic expression of GLAST gene was unaffected by EcoHIV, whereas GLT-1 mRNA was increased by twofold. Conclusions: We provide first evidence that activation of JAK/STAT3 signaling by OSM inhibits glutamate uptake in astrocytes, which results in neuronal excitotoxicity. Our findings with EcoHIV suggest that targeting OSMR-β signaling in astrocytes might alleviate HIV-1-associated excitotoxicity. |
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oai:estudogeral.uc.pt:10316/108666 |
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Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicityGlutamateGLASTGLT-1AstrocytesOncostatin MInterleukin 6NMDAExcitotoxicityHIVAmino Acid Transport System X-AGAnimalsAntineoplastic AgentsAspartic AcidAstrocytesCells, CulturedCerebral CortexCytokinesEmbryo, MammalianExcitatory Amino Acid AgonistsExcitatory Amino Acid Transporter 2Glial Fibrillary Acidic ProteinGlutamic AcidMiceMice, Inbred C57BLN-MethylaspartateNeuronsOncostatin MOncostatin M Receptor beta SubunitRetroviridae Proteins, OncogenicSignal TransductionBackground: Elevated levels of oncostatin M (OSM), an interleukin-6 cytokine family member, have been observed in HIV-1-associated neurocognitive disorders (HAND) and Alzheimer’s disease. However, the function of OSM in these disease conditions is unclear. Since deficient glutamate uptake by astrocytes is instrumental in HAND-associated neurotoxicity, we hypothesized that OSM impairs glutamate uptake in astrocytes and thereby promotes neuronal excitotoxicity. Methods: Primary cultures of mouse cortical astrocytes, neurons, microglia, and BV2 cell line were used. The expression of glutamate transporters (GLAST/EAAT1 and GLT-1/EAAT2) was investigated using real-time PCR and Western blot, and their activity was assessed by measuring 3H-D-aspartate uptake. Neuronal toxicity was measured using the colorimetric MTT (3-(4,5-dimethylthiazol-2-yl-) 2,5-diphenyltetrazolium bromide) assay and immunocytochemistry. A chimeric HIV-1 that infects murine cells (EcoHIV/NL4-3-GFP virus (EcoHIV)) was used to investigate whether the virus induces OSM, OSM receptor (OSMR)-β, glycoprotein 130 (gp130), GLT-1, GLAST (mRNA and protein), and OSM release (ELISA) in cultured BV2 cells, primary microglia, or astrocytes. Statistical analyses of the data were performed using one-way ANOVA (to allow multiple comparisons) and two-tailed Student’s t test. Results: OSM treatment (10 ng/mL) time-dependently reduced GLAST and GLT-1 expression and inhibited 3H-D-aspartate uptake in cultured astrocytes in a concentration-dependent manner, an effect prevented by the Janus kinase (JAK)/signal transducers and activators of transcription (STAT)3 inhibitor AG490. Down-regulation of astrocytic glutamate transport by OSM resulted in NMDA receptor-dependent excitotoxicity in cortical neurons. Infection with EcoHIV induced OSM gene expression and protein release in BV2 cells and microglia, but not in astrocytes. Conversely, EcoHIV caused a fivefold increase in OSMR-β mRNA (but not gp130) and protein in astrocytes, but not in microglia, which did not express OSMR-β protein. Finally, astrocytic expression of GLAST gene was unaffected by EcoHIV, whereas GLT-1 mRNA was increased by twofold. Conclusions: We provide first evidence that activation of JAK/STAT3 signaling by OSM inhibits glutamate uptake in astrocytes, which results in neuronal excitotoxicity. Our findings with EcoHIV suggest that targeting OSMR-β signaling in astrocytes might alleviate HIV-1-associated excitotoxicity.Springer Nature2016-06-10info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/108666http://hdl.handle.net/10316/108666https://doi.org/10.1186/s12974-016-0613-8eng1742-2094Moidunny, ShamsudheenMatos, MarcoWesseling, EvelynBanerjee, SantanuVolsky, David J.Cunha, Rodrigo A.Agostinho, PaulaBoddeke, Hendrikus W.Roy, Sabitainfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-09-07T10:35:23Zoai:estudogeral.uc.pt:10316/108666Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:24:56.898441Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
title |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
spellingShingle |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity Moidunny, Shamsudheen Glutamate GLAST GLT-1 Astrocytes Oncostatin M Interleukin 6 NMDA Excitotoxicity HIV Amino Acid Transport System X-AG Animals Antineoplastic Agents Aspartic Acid Astrocytes Cells, Cultured Cerebral Cortex Cytokines Embryo, Mammalian Excitatory Amino Acid Agonists Excitatory Amino Acid Transporter 2 Glial Fibrillary Acidic Protein Glutamic Acid Mice Mice, Inbred C57BL N-Methylaspartate Neurons Oncostatin M Oncostatin M Receptor beta Subunit Retroviridae Proteins, Oncogenic Signal Transduction |
title_short |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
title_full |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
title_fullStr |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
title_full_unstemmed |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
title_sort |
Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity |
author |
Moidunny, Shamsudheen |
author_facet |
Moidunny, Shamsudheen Matos, Marco Wesseling, Evelyn Banerjee, Santanu Volsky, David J. Cunha, Rodrigo A. Agostinho, Paula Boddeke, Hendrikus W. Roy, Sabita |
author_role |
author |
author2 |
Matos, Marco Wesseling, Evelyn Banerjee, Santanu Volsky, David J. Cunha, Rodrigo A. Agostinho, Paula Boddeke, Hendrikus W. Roy, Sabita |
author2_role |
author author author author author author author author |
dc.contributor.author.fl_str_mv |
Moidunny, Shamsudheen Matos, Marco Wesseling, Evelyn Banerjee, Santanu Volsky, David J. Cunha, Rodrigo A. Agostinho, Paula Boddeke, Hendrikus W. Roy, Sabita |
dc.subject.por.fl_str_mv |
Glutamate GLAST GLT-1 Astrocytes Oncostatin M Interleukin 6 NMDA Excitotoxicity HIV Amino Acid Transport System X-AG Animals Antineoplastic Agents Aspartic Acid Astrocytes Cells, Cultured Cerebral Cortex Cytokines Embryo, Mammalian Excitatory Amino Acid Agonists Excitatory Amino Acid Transporter 2 Glial Fibrillary Acidic Protein Glutamic Acid Mice Mice, Inbred C57BL N-Methylaspartate Neurons Oncostatin M Oncostatin M Receptor beta Subunit Retroviridae Proteins, Oncogenic Signal Transduction |
topic |
Glutamate GLAST GLT-1 Astrocytes Oncostatin M Interleukin 6 NMDA Excitotoxicity HIV Amino Acid Transport System X-AG Animals Antineoplastic Agents Aspartic Acid Astrocytes Cells, Cultured Cerebral Cortex Cytokines Embryo, Mammalian Excitatory Amino Acid Agonists Excitatory Amino Acid Transporter 2 Glial Fibrillary Acidic Protein Glutamic Acid Mice Mice, Inbred C57BL N-Methylaspartate Neurons Oncostatin M Oncostatin M Receptor beta Subunit Retroviridae Proteins, Oncogenic Signal Transduction |
description |
Background: Elevated levels of oncostatin M (OSM), an interleukin-6 cytokine family member, have been observed in HIV-1-associated neurocognitive disorders (HAND) and Alzheimer’s disease. However, the function of OSM in these disease conditions is unclear. Since deficient glutamate uptake by astrocytes is instrumental in HAND-associated neurotoxicity, we hypothesized that OSM impairs glutamate uptake in astrocytes and thereby promotes neuronal excitotoxicity. Methods: Primary cultures of mouse cortical astrocytes, neurons, microglia, and BV2 cell line were used. The expression of glutamate transporters (GLAST/EAAT1 and GLT-1/EAAT2) was investigated using real-time PCR and Western blot, and their activity was assessed by measuring 3H-D-aspartate uptake. Neuronal toxicity was measured using the colorimetric MTT (3-(4,5-dimethylthiazol-2-yl-) 2,5-diphenyltetrazolium bromide) assay and immunocytochemistry. A chimeric HIV-1 that infects murine cells (EcoHIV/NL4-3-GFP virus (EcoHIV)) was used to investigate whether the virus induces OSM, OSM receptor (OSMR)-β, glycoprotein 130 (gp130), GLT-1, GLAST (mRNA and protein), and OSM release (ELISA) in cultured BV2 cells, primary microglia, or astrocytes. Statistical analyses of the data were performed using one-way ANOVA (to allow multiple comparisons) and two-tailed Student’s t test. Results: OSM treatment (10 ng/mL) time-dependently reduced GLAST and GLT-1 expression and inhibited 3H-D-aspartate uptake in cultured astrocytes in a concentration-dependent manner, an effect prevented by the Janus kinase (JAK)/signal transducers and activators of transcription (STAT)3 inhibitor AG490. Down-regulation of astrocytic glutamate transport by OSM resulted in NMDA receptor-dependent excitotoxicity in cortical neurons. Infection with EcoHIV induced OSM gene expression and protein release in BV2 cells and microglia, but not in astrocytes. Conversely, EcoHIV caused a fivefold increase in OSMR-β mRNA (but not gp130) and protein in astrocytes, but not in microglia, which did not express OSMR-β protein. Finally, astrocytic expression of GLAST gene was unaffected by EcoHIV, whereas GLT-1 mRNA was increased by twofold. Conclusions: We provide first evidence that activation of JAK/STAT3 signaling by OSM inhibits glutamate uptake in astrocytes, which results in neuronal excitotoxicity. Our findings with EcoHIV suggest that targeting OSMR-β signaling in astrocytes might alleviate HIV-1-associated excitotoxicity. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-06-10 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/108666 http://hdl.handle.net/10316/108666 https://doi.org/10.1186/s12974-016-0613-8 |
url |
http://hdl.handle.net/10316/108666 https://doi.org/10.1186/s12974-016-0613-8 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1742-2094 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Springer Nature |
publisher.none.fl_str_mv |
Springer Nature |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799134132803993600 |