Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors

In primary cultures of rat cerebellar granule cells with a functional network of glutamatergic neurons, the expression pattern of the different subunits of nitric-oxide (NO)-sensitive guanylyl cyclase changes during cell differentiation. These cells express the alpha1, alpha2 and beta1 subunits of N...

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Detalles Bibliográficos
Autores: Jurado, Sandra, Sánchez-Prieto, José, Torres, Magdalena
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2003
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/307426
Acceso en línea:http://hdl.handle.net/10261/307426
Access Level:acceso abierto
Palabra clave:Cerebellum
N-methyl-D-aspartate receptors
Granule cells
NO-sensitive guanylyl cyclase
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spelling Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptorsJurado, SandraSánchez-Prieto, JoséTorres, MagdalenaCerebellumN-methyl-D-aspartate receptorsGranule cellsNO-sensitive guanylyl cyclaseIn primary cultures of rat cerebellar granule cells with a functional network of glutamatergic neurons, the expression pattern of the different subunits of nitric-oxide (NO)-sensitive guanylyl cyclase changes during cell differentiation. These cells express the alpha1, alpha2 and beta1 subunits of NO-sensitive guanylyl cyclase and synthesize cyclic guanosine monophosphate (cGMP) in response to exogenous or endogenous nitric oxide. In this study, we determined the protein content of the alpha1 and beta1 subunits and quantified alpha1, alpha2 and beta1 mRNA by reverse transcription coupled to a polymerase chain reaction (RT-PCR). Expression of the beta1 subunit increased with the degree of cell differentiation, although most marked changes occurred at the alpha subunit level. In cells freshly isolated from rat pups on postnatal day 7 (P7) the most abundant alpha subunit was alpha1, while alpha2 appeared as the predominant subunit of this type in cultured cells. N-methyl-D-aspartate (NMDA) receptor stimulation in 7- or 14-day-cultured cells led to the upregulation of guanylyl cyclase subunit mRNAs; alpha2 mRNA levels undergoing most significant change. This enhanced subunit expression was accompanied by an increase in the amount of cGMP synthesized in response to NO. Thus, it seems that alpha2 subunits are increasingly expressed as granule cells mature. The presence of this subunit in the guanylyl cyclase heterodimer facilitates its localization at synaptic membranes, where the enzyme acts as a sensor for NO formed by the postsynaptic protein 95 (PSD-95)-associated neuronal NO synthase.This study was funded by a grant from the Ministerio de Ciencia y Tecnología (PM99-0058). Sandra Jurado was supported by a fellowship from the same Ministry.Peer reviewedCompany of BiologistsMinisterio de Ciencia y Tecnología (España)202320232003info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/307426reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésJournal of Cell Sciencehttps://doi.org/10.1242/jcs.00620Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3074262026-05-22T06:33:51Z
dc.title.none.fl_str_mv Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
title Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
spellingShingle Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
Jurado, Sandra
Cerebellum
N-methyl-D-aspartate receptors
Granule cells
NO-sensitive guanylyl cyclase
title_short Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
title_full Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
title_fullStr Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
title_full_unstemmed Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
title_sort Differential expression of NO-sensitive guanylyl cyclase subunits during the development of rat cerebellar granule cells: regulation via N-methyl-Daspartate receptors
dc.creator.none.fl_str_mv Jurado, Sandra
Sánchez-Prieto, José
Torres, Magdalena
author Jurado, Sandra
author_facet Jurado, Sandra
Sánchez-Prieto, José
Torres, Magdalena
author_role author
author2 Sánchez-Prieto, José
Torres, Magdalena
author2_role author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia y Tecnología (España)
dc.subject.none.fl_str_mv Cerebellum
N-methyl-D-aspartate receptors
Granule cells
NO-sensitive guanylyl cyclase
topic Cerebellum
N-methyl-D-aspartate receptors
Granule cells
NO-sensitive guanylyl cyclase
description In primary cultures of rat cerebellar granule cells with a functional network of glutamatergic neurons, the expression pattern of the different subunits of nitric-oxide (NO)-sensitive guanylyl cyclase changes during cell differentiation. These cells express the alpha1, alpha2 and beta1 subunits of NO-sensitive guanylyl cyclase and synthesize cyclic guanosine monophosphate (cGMP) in response to exogenous or endogenous nitric oxide. In this study, we determined the protein content of the alpha1 and beta1 subunits and quantified alpha1, alpha2 and beta1 mRNA by reverse transcription coupled to a polymerase chain reaction (RT-PCR). Expression of the beta1 subunit increased with the degree of cell differentiation, although most marked changes occurred at the alpha subunit level. In cells freshly isolated from rat pups on postnatal day 7 (P7) the most abundant alpha subunit was alpha1, while alpha2 appeared as the predominant subunit of this type in cultured cells. N-methyl-D-aspartate (NMDA) receptor stimulation in 7- or 14-day-cultured cells led to the upregulation of guanylyl cyclase subunit mRNAs; alpha2 mRNA levels undergoing most significant change. This enhanced subunit expression was accompanied by an increase in the amount of cGMP synthesized in response to NO. Thus, it seems that alpha2 subunits are increasingly expressed as granule cells mature. The presence of this subunit in the guanylyl cyclase heterodimer facilitates its localization at synaptic membranes, where the enzyme acts as a sensor for NO formed by the postsynaptic protein 95 (PSD-95)-associated neuronal NO synthase.
publishDate 2003
dc.date.none.fl_str_mv 2003
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/307426
url http://hdl.handle.net/10261/307426
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Journal of Cell Science
https://doi.org/10.1242/jcs.00620
No
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Company of Biologists
publisher.none.fl_str_mv Company of Biologists
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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