PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing

Autores
Gentili, Claudia Rosana; Boland, Ricardo Leopoldo; Russo, Ana Josefa
Año de publicación
2001
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We previously reported that in rat duodenal cells (enterocytes), parathyroid hormone (PTH [1–34]: PTH) stimulates the hydrolysis of polyphosphoinositides by phospholipase C (PLC), generating the second messengers inositol trisphosphate (IP3) and diacylglycerol (DAG) and that this mechanism is severely altered in old animals. In the present study, we show that PTH [1–34]-dependent IP3 release in young rats was blocked to a great extent by an antibody against guanine nucleotide binding protein Gαq/11, indicating that the hormone activates a β isoform of PLC coupled to the α subunit of Gq/11. In addition, PTH rapidly (within 30 s, with maximal effects at 1 min) stimulated tyrosine phosphorylation of PLCγ in a dose-dependent fashion (10−10–10−7 M). The hormone response was specific as PTH [7–34] was without effects. The tyrosine kinase inhibitors, genistein (100 μM) and herbimycin (2 μM), suppressed PTH-dependent PLCγ tyrosine phosphorylation. Stimulation of PLCγ tyrosine phosphorylation by PTH [1–34] greatly decreased with ageing. PP1 (10 μM), a specific inhibitor of the Src family of tyrosine kinases, completely abolished PLCγ phosphorylation. The hormone-induced Src tyrosine dephosphorylation, a major mechanism of Src activation, an effect that was blunted in old animals. These results indicate that in rat enterocytes PTH generates IP3 mainly through G-protein-coupled PLCβ and stimulates PLCγ phosphorylation via the nonreceptor tyrosine kinase Src. Impairment of PTH activation of both PLC isoforms upon ageing may result in abnormal hormone regulation of cell Ca2+ and proliferation in the duodenum.
Fil: Gentili, Claudia Rosana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Materia
Pth
Rat Enterocytes
Ip3
Plcβ
Plcγ
Ageing
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:11336/40351
Acceder
id CONICETDig_f1b2bbc1295f466fe8c87aabf926a609
oai_identifier_str oai:ri.conicet.gov.ar:11336/40351
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageingGentili, Claudia RosanaBoland, Ricardo LeopoldoRusso, Ana JosefaPthRat EnterocytesIp3PlcβPlcγAgeinghttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We previously reported that in rat duodenal cells (enterocytes), parathyroid hormone (PTH [1–34]: PTH) stimulates the hydrolysis of polyphosphoinositides by phospholipase C (PLC), generating the second messengers inositol trisphosphate (IP3) and diacylglycerol (DAG) and that this mechanism is severely altered in old animals. In the present study, we show that PTH [1–34]-dependent IP3 release in young rats was blocked to a great extent by an antibody against guanine nucleotide binding protein Gαq/11, indicating that the hormone activates a β isoform of PLC coupled to the α subunit of Gq/11. In addition, PTH rapidly (within 30 s, with maximal effects at 1 min) stimulated tyrosine phosphorylation of PLCγ in a dose-dependent fashion (10−10–10−7 M). The hormone response was specific as PTH [7–34] was without effects. The tyrosine kinase inhibitors, genistein (100 μM) and herbimycin (2 μM), suppressed PTH-dependent PLCγ tyrosine phosphorylation. Stimulation of PLCγ tyrosine phosphorylation by PTH [1–34] greatly decreased with ageing. PP1 (10 μM), a specific inhibitor of the Src family of tyrosine kinases, completely abolished PLCγ phosphorylation. The hormone-induced Src tyrosine dephosphorylation, a major mechanism of Src activation, an effect that was blunted in old animals. These results indicate that in rat enterocytes PTH generates IP3 mainly through G-protein-coupled PLCβ and stimulates PLCγ phosphorylation via the nonreceptor tyrosine kinase Src. Impairment of PTH activation of both PLC isoforms upon ageing may result in abnormal hormone regulation of cell Ca2+ and proliferation in the duodenum.Fil: Gentili, Claudia Rosana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; ArgentinaFil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaElsevier Science Inc2001-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/40351Gentili, Claudia Rosana; Boland, Ricardo Leopoldo; Russo, Ana Josefa; PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing; Elsevier Science Inc; Cellular Signalling; 13; 2; 2-2001; 131-1380898-6568CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/S0898-6568(00)00145-5info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0898656800001455info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:54:04Zoai:ri.conicet.gov.ar:11336/40351instacron: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 09:54:04.634CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
title PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
spellingShingle PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
Gentili, Claudia Rosana
Pth
Rat Enterocytes
Ip3
Plcβ
Plcγ
Ageing
title_short PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
title_full PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
title_fullStr PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
title_full_unstemmed PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
title_sort PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing
dc.creator.none.fl_str_mv Gentili, Claudia Rosana
Boland, Ricardo Leopoldo
Russo, Ana Josefa
author Gentili, Claudia Rosana
author_facet Gentili, Claudia Rosana
Boland, Ricardo Leopoldo
Russo, Ana Josefa
author_role author
author2 Boland, Ricardo Leopoldo
Russo, Ana Josefa
author2_role author
author
dc.subject.none.fl_str_mv Pth
Rat Enterocytes
Ip3
Plcβ
Plcγ
Ageing
topic Pth
Rat Enterocytes
Ip3
Plcβ
Plcγ
Ageing
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We previously reported that in rat duodenal cells (enterocytes), parathyroid hormone (PTH [1–34]: PTH) stimulates the hydrolysis of polyphosphoinositides by phospholipase C (PLC), generating the second messengers inositol trisphosphate (IP3) and diacylglycerol (DAG) and that this mechanism is severely altered in old animals. In the present study, we show that PTH [1–34]-dependent IP3 release in young rats was blocked to a great extent by an antibody against guanine nucleotide binding protein Gαq/11, indicating that the hormone activates a β isoform of PLC coupled to the α subunit of Gq/11. In addition, PTH rapidly (within 30 s, with maximal effects at 1 min) stimulated tyrosine phosphorylation of PLCγ in a dose-dependent fashion (10−10–10−7 M). The hormone response was specific as PTH [7–34] was without effects. The tyrosine kinase inhibitors, genistein (100 μM) and herbimycin (2 μM), suppressed PTH-dependent PLCγ tyrosine phosphorylation. Stimulation of PLCγ tyrosine phosphorylation by PTH [1–34] greatly decreased with ageing. PP1 (10 μM), a specific inhibitor of the Src family of tyrosine kinases, completely abolished PLCγ phosphorylation. The hormone-induced Src tyrosine dephosphorylation, a major mechanism of Src activation, an effect that was blunted in old animals. These results indicate that in rat enterocytes PTH generates IP3 mainly through G-protein-coupled PLCβ and stimulates PLCγ phosphorylation via the nonreceptor tyrosine kinase Src. Impairment of PTH activation of both PLC isoforms upon ageing may result in abnormal hormone regulation of cell Ca2+ and proliferation in the duodenum.
Fil: Gentili, Claudia Rosana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
description We previously reported that in rat duodenal cells (enterocytes), parathyroid hormone (PTH [1–34]: PTH) stimulates the hydrolysis of polyphosphoinositides by phospholipase C (PLC), generating the second messengers inositol trisphosphate (IP3) and diacylglycerol (DAG) and that this mechanism is severely altered in old animals. In the present study, we show that PTH [1–34]-dependent IP3 release in young rats was blocked to a great extent by an antibody against guanine nucleotide binding protein Gαq/11, indicating that the hormone activates a β isoform of PLC coupled to the α subunit of Gq/11. In addition, PTH rapidly (within 30 s, with maximal effects at 1 min) stimulated tyrosine phosphorylation of PLCγ in a dose-dependent fashion (10−10–10−7 M). The hormone response was specific as PTH [7–34] was without effects. The tyrosine kinase inhibitors, genistein (100 μM) and herbimycin (2 μM), suppressed PTH-dependent PLCγ tyrosine phosphorylation. Stimulation of PLCγ tyrosine phosphorylation by PTH [1–34] greatly decreased with ageing. PP1 (10 μM), a specific inhibitor of the Src family of tyrosine kinases, completely abolished PLCγ phosphorylation. The hormone-induced Src tyrosine dephosphorylation, a major mechanism of Src activation, an effect that was blunted in old animals. These results indicate that in rat enterocytes PTH generates IP3 mainly through G-protein-coupled PLCβ and stimulates PLCγ phosphorylation via the nonreceptor tyrosine kinase Src. Impairment of PTH activation of both PLC isoforms upon ageing may result in abnormal hormone regulation of cell Ca2+ and proliferation in the duodenum.
publishDate 2001
dc.date.none.fl_str_mv 2001-02
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/11336/40351
Gentili, Claudia Rosana; Boland, Ricardo Leopoldo; Russo, Ana Josefa; PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing; Elsevier Science Inc; Cellular Signalling; 13; 2; 2-2001; 131-138
0898-6568
CONICET Digital
CONICET
url http://hdl.handle.net/11336/40351
identifier_str_mv Gentili, Claudia Rosana; Boland, Ricardo Leopoldo; Russo, Ana Josefa; PTH stimulates PLCβ and PLCγ isoenzymes in rat enterocytes: influence of ageing; Elsevier Science Inc; Cellular Signalling; 13; 2; 2-2001; 131-138
0898-6568
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/S0898-6568(00)00145-5
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0898656800001455
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
dc.publisher.none.fl_str_mv Elsevier Science Inc
publisher.none.fl_str_mv Elsevier Science Inc
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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