Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury
Autor(a) principal: | |
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Data de Publicação: | 1999 |
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/10316/8510 https://doi.org/10.1023/A:1020977331372 |
Resumo: | In this study, we analyzed how distinct glycolysis inhibitors influenced the redox status of retinal cells, used as a neuronal model. Three different approaches were used to inhibit glycolysis: the cells were submitted to iodoacetic acid (IAA), an inhibitor of glyceraldehyde 3-phosphate dehydrogenase, to 2-deoxy-glucose (DG) in glucose-free medium, which was used as a substitute of glucose, or in the absence of glucose. The redox status of the cells was evaluated by determining the reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). By the analysis of dose-response curves of MTT reduction, IAA showed values of IC50 = 7.02 × 10-5 M, whereas DG showed values of IC50 = 7.42 × 10-4 M. Upon 30 min-incubation, glucose deprivation, per se, did not significantly affect MTT reduction. We also evaluated the reduction of MTT as an indicator of cell injury by exposing the cells to 100 µM glutamate during the decrement of glycolysis function. In the presence of glutamate, for 2 h, there was a decrease in MTT reduction, which was potentiated in the presence of DG (10-20% decrease), in the presence of IAA (about 30% decrease) or in glucose-free medium (about 30% decrease). Major changes observed by the MTT assay, upon exposure to glutamate, indicative of changes in the redox status of retinal cells, were concomitant with variations in intracellular ATP. Under glucose deprivation, endogenous ATP decreased significantly from 38.9 ± 4.4 to 13.3 ± 0.7 nmol/mg protein after exposure to 100 µM glutamate. The results support a different vulnerability of retinal cells after being exposed to distinct forms of glycolysis inhibition. |
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Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated InjuryIn this study, we analyzed how distinct glycolysis inhibitors influenced the redox status of retinal cells, used as a neuronal model. Three different approaches were used to inhibit glycolysis: the cells were submitted to iodoacetic acid (IAA), an inhibitor of glyceraldehyde 3-phosphate dehydrogenase, to 2-deoxy-glucose (DG) in glucose-free medium, which was used as a substitute of glucose, or in the absence of glucose. The redox status of the cells was evaluated by determining the reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). By the analysis of dose-response curves of MTT reduction, IAA showed values of IC50 = 7.02 × 10-5 M, whereas DG showed values of IC50 = 7.42 × 10-4 M. Upon 30 min-incubation, glucose deprivation, per se, did not significantly affect MTT reduction. We also evaluated the reduction of MTT as an indicator of cell injury by exposing the cells to 100 µM glutamate during the decrement of glycolysis function. In the presence of glutamate, for 2 h, there was a decrease in MTT reduction, which was potentiated in the presence of DG (10-20% decrease), in the presence of IAA (about 30% decrease) or in glucose-free medium (about 30% decrease). Major changes observed by the MTT assay, upon exposure to glutamate, indicative of changes in the redox status of retinal cells, were concomitant with variations in intracellular ATP. Under glucose deprivation, endogenous ATP decreased significantly from 38.9 ± 4.4 to 13.3 ± 0.7 nmol/mg protein after exposure to 100 µM glutamate. The results support a different vulnerability of retinal cells after being exposed to distinct forms of glycolysis inhibition.1999info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/8510http://hdl.handle.net/10316/8510https://doi.org/10.1023/A:1020977331372engNeurochemical Research. 24:3 (1999) 351-358Rego, Ana CristinaAreias, FilipeSantos, Maria SanchaOliveira, Catarinainfo: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:RCAAP2020-05-27T16:14:34Zoai:estudogeral.uc.pt:10316/8510Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:43:33.959156Repositório 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 |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
title |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
spellingShingle |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury Rego, Ana Cristina |
title_short |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
title_full |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
title_fullStr |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
title_full_unstemmed |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
title_sort |
Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury |
author |
Rego, Ana Cristina |
author_facet |
Rego, Ana Cristina Areias, Filipe Santos, Maria Sancha Oliveira, Catarina |
author_role |
author |
author2 |
Areias, Filipe Santos, Maria Sancha Oliveira, Catarina |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Rego, Ana Cristina Areias, Filipe Santos, Maria Sancha Oliveira, Catarina |
description |
In this study, we analyzed how distinct glycolysis inhibitors influenced the redox status of retinal cells, used as a neuronal model. Three different approaches were used to inhibit glycolysis: the cells were submitted to iodoacetic acid (IAA), an inhibitor of glyceraldehyde 3-phosphate dehydrogenase, to 2-deoxy-glucose (DG) in glucose-free medium, which was used as a substitute of glucose, or in the absence of glucose. The redox status of the cells was evaluated by determining the reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). By the analysis of dose-response curves of MTT reduction, IAA showed values of IC50 = 7.02 × 10-5 M, whereas DG showed values of IC50 = 7.42 × 10-4 M. Upon 30 min-incubation, glucose deprivation, per se, did not significantly affect MTT reduction. We also evaluated the reduction of MTT as an indicator of cell injury by exposing the cells to 100 µM glutamate during the decrement of glycolysis function. In the presence of glutamate, for 2 h, there was a decrease in MTT reduction, which was potentiated in the presence of DG (10-20% decrease), in the presence of IAA (about 30% decrease) or in glucose-free medium (about 30% decrease). Major changes observed by the MTT assay, upon exposure to glutamate, indicative of changes in the redox status of retinal cells, were concomitant with variations in intracellular ATP. Under glucose deprivation, endogenous ATP decreased significantly from 38.9 ± 4.4 to 13.3 ± 0.7 nmol/mg protein after exposure to 100 µM glutamate. The results support a different vulnerability of retinal cells after being exposed to distinct forms of glycolysis inhibition. |
publishDate |
1999 |
dc.date.none.fl_str_mv |
1999 |
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/10316/8510 http://hdl.handle.net/10316/8510 https://doi.org/10.1023/A:1020977331372 |
url |
http://hdl.handle.net/10316/8510 https://doi.org/10.1023/A:1020977331372 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Neurochemical Research. 24:3 (1999) 351-358 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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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) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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