Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit
Autor(a) principal: | |
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Data de Publicação: | 2003 |
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/10316/5408 https://doi.org/10.1016/S0165-3806(02)00580-1 |
Resumo: | The functionality of [alpha]-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors in chick embryo retina cells during development in vitro was studied by using Co2+ uptake, and these data were correlated with the expression of the AMPA receptor subunit GluR4. We found that at 5 h in vitro only a small number of cells took up Co2+ upon stimulation with 100 [mu]M kainate or other AMPA receptor agonists, in the presence of cyclothiazide (CTZ), to inhibit desensitisation. The number of cells sensitive to kainate increased from 5 h in vitro to 3 days in vitro (DIV), and remained relatively constant until 14 DIV. When the cells were stimulated with (2S,4S)-4-methylglutamic acid (30 [mu]M), a specific kainate receptor agonist, after inhibiting desensitisation with concanavalin A, we did not observe an increase in the number of cells responding, as compared to the control. The expression of the AMPA receptor subunit GluR4 during development was detected by immunofluorescence mainly at the perinuclear region of the cells, and the number of positive cells increased from 5 h in vitro to 7 DIV, and remained relatively constant until 14 DIV. The results suggest that AMPA receptors can be functionally active at early embryonic stages (5 h in vitro) in cultured retinal neurons, although in only a few cells, before synapse formation (E12). The localisation of GluR4 was well correlated with Co2+ entry, since the strongest GluR4 immunoreactivity was found in the regions that showed the most intense labelling with Co2+. Finally, we found that the expression levels of GluR4 at the neurites increased between 5 h in vitro and 7 DIV, near the period of synapse formation. |
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Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunitAMPA receptorDevelopmentNon-NMDA receptorThe functionality of [alpha]-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors in chick embryo retina cells during development in vitro was studied by using Co2+ uptake, and these data were correlated with the expression of the AMPA receptor subunit GluR4. We found that at 5 h in vitro only a small number of cells took up Co2+ upon stimulation with 100 [mu]M kainate or other AMPA receptor agonists, in the presence of cyclothiazide (CTZ), to inhibit desensitisation. The number of cells sensitive to kainate increased from 5 h in vitro to 3 days in vitro (DIV), and remained relatively constant until 14 DIV. When the cells were stimulated with (2S,4S)-4-methylglutamic acid (30 [mu]M), a specific kainate receptor agonist, after inhibiting desensitisation with concanavalin A, we did not observe an increase in the number of cells responding, as compared to the control. The expression of the AMPA receptor subunit GluR4 during development was detected by immunofluorescence mainly at the perinuclear region of the cells, and the number of positive cells increased from 5 h in vitro to 7 DIV, and remained relatively constant until 14 DIV. The results suggest that AMPA receptors can be functionally active at early embryonic stages (5 h in vitro) in cultured retinal neurons, although in only a few cells, before synapse formation (E12). The localisation of GluR4 was well correlated with Co2+ entry, since the strongest GluR4 immunoreactivity was found in the regions that showed the most intense labelling with Co2+. Finally, we found that the expression levels of GluR4 at the neurites increased between 5 h in vitro and 7 DIV, near the period of synapse formation.http://www.sciencedirect.com/science/article/B6SYW-47CYB0J-2/1/56187dd630d63c1a50293ba3010eff932003info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleaplication/PDFhttp://hdl.handle.net/10316/5408http://hdl.handle.net/10316/5408https://doi.org/10.1016/S0165-3806(02)00580-1engDevelopmental Brain Research. 140:1 (2003) 75-84Cristóvão, Armando J.Carvalho, Caetana M.info: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:RCAAP2020-11-06T16:59:37Zoai:estudogeral.uc.pt:10316/5408Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:55:28.965918Repositório 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 |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
title |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
spellingShingle |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit Cristóvão, Armando J. AMPA receptor Development Non-NMDA receptor |
title_short |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
title_full |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
title_fullStr |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
title_full_unstemmed |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
title_sort |
Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit |
author |
Cristóvão, Armando J. |
author_facet |
Cristóvão, Armando J. Carvalho, Caetana M. |
author_role |
author |
author2 |
Carvalho, Caetana M. |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Cristóvão, Armando J. Carvalho, Caetana M. |
dc.subject.por.fl_str_mv |
AMPA receptor Development Non-NMDA receptor |
topic |
AMPA receptor Development Non-NMDA receptor |
description |
The functionality of [alpha]-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors in chick embryo retina cells during development in vitro was studied by using Co2+ uptake, and these data were correlated with the expression of the AMPA receptor subunit GluR4. We found that at 5 h in vitro only a small number of cells took up Co2+ upon stimulation with 100 [mu]M kainate or other AMPA receptor agonists, in the presence of cyclothiazide (CTZ), to inhibit desensitisation. The number of cells sensitive to kainate increased from 5 h in vitro to 3 days in vitro (DIV), and remained relatively constant until 14 DIV. When the cells were stimulated with (2S,4S)-4-methylglutamic acid (30 [mu]M), a specific kainate receptor agonist, after inhibiting desensitisation with concanavalin A, we did not observe an increase in the number of cells responding, as compared to the control. The expression of the AMPA receptor subunit GluR4 during development was detected by immunofluorescence mainly at the perinuclear region of the cells, and the number of positive cells increased from 5 h in vitro to 7 DIV, and remained relatively constant until 14 DIV. The results suggest that AMPA receptors can be functionally active at early embryonic stages (5 h in vitro) in cultured retinal neurons, although in only a few cells, before synapse formation (E12). The localisation of GluR4 was well correlated with Co2+ entry, since the strongest GluR4 immunoreactivity was found in the regions that showed the most intense labelling with Co2+. Finally, we found that the expression levels of GluR4 at the neurites increased between 5 h in vitro and 7 DIV, near the period of synapse formation. |
publishDate |
2003 |
dc.date.none.fl_str_mv |
2003 |
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/10316/5408 http://hdl.handle.net/10316/5408 https://doi.org/10.1016/S0165-3806(02)00580-1 |
url |
http://hdl.handle.net/10316/5408 https://doi.org/10.1016/S0165-3806(02)00580-1 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Developmental Brain Research. 140:1 (2003) 75-84 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
aplication/PDF |
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 |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
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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1799133842122997760 |