Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells

Autores
Cymeryng, Cora Betriz; Lotito, Sebastian; Colonna, Cecilia; Finkielstein, Carla; Pomeraniec, Yael; Grión, Natalia; Gadda, Luciana; Maloberti, Paula Mariana; Podesta, Ernesto Jorge
Año de publicación
2002
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Nitric oxide (NO) synthase (NOS) expression was analyzed in rat adrenal zona fasciculata. Both neuronal NOS and endothelial NOS mRNAs were detected by RT-PCR, immunohistochemistry, and immunoblot analysis. The biochemical characterization of adrenal zona fasciculata NOS enzymatic activity confirmed the presence of a constitutive isoform. In a cell line derived from mouse adrenal cortex, only endothelial NOS expression was detected by both RT-PCR and immunoblot analysis. Nitrate plus nitrite levels in Y1 cell incubation medium were increased in the presence of L-arginine and the calcium ionophore A23187, but not D-arginine, indicating enzymatic activity. Moreover, a low, but significant, conversion of Larginine to L-citrulline, abolished by the NOS inhibitor, N(G)-nitro-L-arginine, was detected in Y1 cells. The effect of L-arginine on pregnenolone production was examined. L-Arginine decreased both basal and ACTH-stimulated pregnenolone production in Y1 cells. The inhibitory effect of L-arginine could be attributed to endogenously generated NO, because it was blocked by N(G)-nitro-L-arginine, and it was mimicked by the addition of a NO donor, diethylenetriamine-NO. An inhibitory effect of NO on pregnenolone production from 22Rhydroxycholesterol and on steroidogenic acute regulatory protein expression was also determined. Taken together, these results suggest that at least part of the adrenal NO could derive from steroidogenic cells and modulate their function.
Fil: Cymeryng, Cora Betriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Lotito, Sebastian. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquimica Humana; Argentina
Fil: Colonna, Cecilia. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquímica Humana; Argentina
Fil: Finkielstein, Carla. Universidad de Buenos Aires.Facultad de Medicina.Departamento de Bioquímica Humana; Argentina
Fil: Pomeraniec, Yael. Universidad de Buenos Aires; Argentina
Fil: Grión, Natalia. Universidad de Buenos Aires; Argentina
Fil: Gadda, Luciana. Universidad de Buenos Aires; Argentina
Fil: Maloberti, Paula Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; Argentina
Fil: Podesta, Ernesto Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; Argentina
Materia
Nitric oxide
RT-PCR
mRNAs
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/123215
Acceder
id CONICETDig_b6fe6fdf2ce2977af4409cc73c6bf044
oai_identifier_str oai:ri.conicet.gov.ar:11336/123215
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata CellsCymeryng, Cora BetrizLotito, SebastianColonna, CeciliaFinkielstein, CarlaPomeraniec, YaelGrión, NataliaGadda, LucianaMaloberti, Paula MarianaPodesta, Ernesto JorgeNitric oxideRT-PCRmRNAshttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Nitric oxide (NO) synthase (NOS) expression was analyzed in rat adrenal zona fasciculata. Both neuronal NOS and endothelial NOS mRNAs were detected by RT-PCR, immunohistochemistry, and immunoblot analysis. The biochemical characterization of adrenal zona fasciculata NOS enzymatic activity confirmed the presence of a constitutive isoform. In a cell line derived from mouse adrenal cortex, only endothelial NOS expression was detected by both RT-PCR and immunoblot analysis. Nitrate plus nitrite levels in Y1 cell incubation medium were increased in the presence of L-arginine and the calcium ionophore A23187, but not D-arginine, indicating enzymatic activity. Moreover, a low, but significant, conversion of Larginine to L-citrulline, abolished by the NOS inhibitor, N(G)-nitro-L-arginine, was detected in Y1 cells. The effect of L-arginine on pregnenolone production was examined. L-Arginine decreased both basal and ACTH-stimulated pregnenolone production in Y1 cells. The inhibitory effect of L-arginine could be attributed to endogenously generated NO, because it was blocked by N(G)-nitro-L-arginine, and it was mimicked by the addition of a NO donor, diethylenetriamine-NO. An inhibitory effect of NO on pregnenolone production from 22Rhydroxycholesterol and on steroidogenic acute regulatory protein expression was also determined. Taken together, these results suggest that at least part of the adrenal NO could derive from steroidogenic cells and modulate their function.Fil: Cymeryng, Cora Betriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Lotito, Sebastian. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquimica Humana; ArgentinaFil: Colonna, Cecilia. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquímica Humana; ArgentinaFil: Finkielstein, Carla. Universidad de Buenos Aires.Facultad de Medicina.Departamento de Bioquímica Humana; ArgentinaFil: Pomeraniec, Yael. Universidad de Buenos Aires; ArgentinaFil: Grión, Natalia. Universidad de Buenos Aires; ArgentinaFil: Gadda, Luciana. Universidad de Buenos Aires; ArgentinaFil: Maloberti, Paula Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Podesta, Ernesto Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaEndocrine Society2002-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/123215Cymeryng, Cora Betriz; Lotito, Sebastian; Colonna, Cecilia; Finkielstein, Carla; Pomeraniec, Yael; et al.; Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells; Endocrine Society; Endocrinology; 143; 4; 4-2002; 1235-12420013-7227CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/endo/article/143/4/1235/2989228info:eu-repo/semantics/altIdentifier/doi/10.1210/endo.143.4.8727info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:48:13Zoai:ri.conicet.gov.ar:11336/123215instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:48:13.798CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
title Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
spellingShingle Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
Cymeryng, Cora Betriz
Nitric oxide
RT-PCR
mRNAs
title_short Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
title_full Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
title_fullStr Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
title_full_unstemmed Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
title_sort Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells
dc.creator.none.fl_str_mv Cymeryng, Cora Betriz
Lotito, Sebastian
Colonna, Cecilia
Finkielstein, Carla
Pomeraniec, Yael
Grión, Natalia
Gadda, Luciana
Maloberti, Paula Mariana
Podesta, Ernesto Jorge
author Cymeryng, Cora Betriz
author_facet Cymeryng, Cora Betriz
Lotito, Sebastian
Colonna, Cecilia
Finkielstein, Carla
Pomeraniec, Yael
Grión, Natalia
Gadda, Luciana
Maloberti, Paula Mariana
Podesta, Ernesto Jorge
author_role author
author2 Lotito, Sebastian
Colonna, Cecilia
Finkielstein, Carla
Pomeraniec, Yael
Grión, Natalia
Gadda, Luciana
Maloberti, Paula Mariana
Podesta, Ernesto Jorge
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Nitric oxide
RT-PCR
mRNAs
topic Nitric oxide
RT-PCR
mRNAs
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Nitric oxide (NO) synthase (NOS) expression was analyzed in rat adrenal zona fasciculata. Both neuronal NOS and endothelial NOS mRNAs were detected by RT-PCR, immunohistochemistry, and immunoblot analysis. The biochemical characterization of adrenal zona fasciculata NOS enzymatic activity confirmed the presence of a constitutive isoform. In a cell line derived from mouse adrenal cortex, only endothelial NOS expression was detected by both RT-PCR and immunoblot analysis. Nitrate plus nitrite levels in Y1 cell incubation medium were increased in the presence of L-arginine and the calcium ionophore A23187, but not D-arginine, indicating enzymatic activity. Moreover, a low, but significant, conversion of Larginine to L-citrulline, abolished by the NOS inhibitor, N(G)-nitro-L-arginine, was detected in Y1 cells. The effect of L-arginine on pregnenolone production was examined. L-Arginine decreased both basal and ACTH-stimulated pregnenolone production in Y1 cells. The inhibitory effect of L-arginine could be attributed to endogenously generated NO, because it was blocked by N(G)-nitro-L-arginine, and it was mimicked by the addition of a NO donor, diethylenetriamine-NO. An inhibitory effect of NO on pregnenolone production from 22Rhydroxycholesterol and on steroidogenic acute regulatory protein expression was also determined. Taken together, these results suggest that at least part of the adrenal NO could derive from steroidogenic cells and modulate their function.
Fil: Cymeryng, Cora Betriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Lotito, Sebastian. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquimica Humana; Argentina
Fil: Colonna, Cecilia. Universidad de Buenos Aires, Facultad de Medicina,Departamento de Bioquímica Humana; Argentina
Fil: Finkielstein, Carla. Universidad de Buenos Aires.Facultad de Medicina.Departamento de Bioquímica Humana; Argentina
Fil: Pomeraniec, Yael. Universidad de Buenos Aires; Argentina
Fil: Grión, Natalia. Universidad de Buenos Aires; Argentina
Fil: Gadda, Luciana. Universidad de Buenos Aires; Argentina
Fil: Maloberti, Paula Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; Argentina
Fil: Podesta, Ernesto Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; Argentina
description Nitric oxide (NO) synthase (NOS) expression was analyzed in rat adrenal zona fasciculata. Both neuronal NOS and endothelial NOS mRNAs were detected by RT-PCR, immunohistochemistry, and immunoblot analysis. The biochemical characterization of adrenal zona fasciculata NOS enzymatic activity confirmed the presence of a constitutive isoform. In a cell line derived from mouse adrenal cortex, only endothelial NOS expression was detected by both RT-PCR and immunoblot analysis. Nitrate plus nitrite levels in Y1 cell incubation medium were increased in the presence of L-arginine and the calcium ionophore A23187, but not D-arginine, indicating enzymatic activity. Moreover, a low, but significant, conversion of Larginine to L-citrulline, abolished by the NOS inhibitor, N(G)-nitro-L-arginine, was detected in Y1 cells. The effect of L-arginine on pregnenolone production was examined. L-Arginine decreased both basal and ACTH-stimulated pregnenolone production in Y1 cells. The inhibitory effect of L-arginine could be attributed to endogenously generated NO, because it was blocked by N(G)-nitro-L-arginine, and it was mimicked by the addition of a NO donor, diethylenetriamine-NO. An inhibitory effect of NO on pregnenolone production from 22Rhydroxycholesterol and on steroidogenic acute regulatory protein expression was also determined. Taken together, these results suggest that at least part of the adrenal NO could derive from steroidogenic cells and modulate their function.
publishDate 2002
dc.date.none.fl_str_mv 2002-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/123215
Cymeryng, Cora Betriz; Lotito, Sebastian; Colonna, Cecilia; Finkielstein, Carla; Pomeraniec, Yael; et al.; Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells; Endocrine Society; Endocrinology; 143; 4; 4-2002; 1235-1242
0013-7227
CONICET Digital
CONICET
url http://hdl.handle.net/11336/123215
identifier_str_mv Cymeryng, Cora Betriz; Lotito, Sebastian; Colonna, Cecilia; Finkielstein, Carla; Pomeraniec, Yael; et al.; Expression of Nitric Oxide Synthases in Rat Adrenal Zona Fasciculata Cells; Endocrine Society; Endocrinology; 143; 4; 4-2002; 1235-1242
0013-7227
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/endo/article/143/4/1235/2989228
info:eu-repo/semantics/altIdentifier/doi/10.1210/endo.143.4.8727
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Endocrine Society
publisher.none.fl_str_mv Endocrine Society
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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