Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells

Autores
Raychowdhury, Malay K.; Ramos, Arnolt J.; Zhang, Peng; McLaughin, Margaret; Dai, Xiao-Qing; Chen, Xing-Zhen; Montalbetti, Nicolas; Cantero, Maria del Rocio; Ausiello, Dennis A.; Cantiello, Horacio Fabio
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The primary cilium of renal epithelial cells is a nonmotile sensory organelle, implicated in mechanosensory transduction signals. Recent studies from our laboratory indicate that renal epithelial primary cilia display abundant channel activity; however, the presence and functional role of specific membrane receptors in this organelle are heretofore unknown. Here, we determined a functional signaling pathway associated with the type 2 vasopressin receptor (V2R) in primary cilia of renal epithelial cells. Besides their normal localization on basolateral membrane, V2R was expressed in primary cilia of LLC-PK1 renal epithelial cells. The presence of V2R in primary cilia was determined by spontaneous fluorescence of a V2R-gfp chimera and confirmed by immunocytochemical analysis of wild-type LLC-PK1 cells stained with anti-V2R antibodies and in LLC-PK1 cells overexpressing the V2R-Flag, with anti-Flag antibody. Ciliary V2R colocalized with adenylyl cyclase (AC) type V/VI in all cell types tested. Functional coupling of the receptors with AC was confirmed by measurement of cAMP production in isolated cilia and by testing AVP-induced cation-selective channel activity either in reconstituted lipid bilayers or subjected to membrane-attached patch clamping. Addition of either 10 μM AVP (trans) or forskolin (cis) in the presence but not the absence of ATP (1 mM, cis) stimulated cation-selective channel activity in ciliary membranes. This channel activity was reduced by addition of the PKA inhibitor PKI. The data provide the first demonstration for the presence of V2R in primary cilia of renal epithelial cells, and a functional cAMP-signaling pathway, which targets ciliary channel function and may help control the sensory function of the primary cilium.
Fil: Raychowdhury, Malay K.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Ramos, Arnolt J.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Zhang, Peng. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: McLaughin, Margaret. Harvard Medical School; Estados Unidos. Massachusetts General Hospital; Estados Unidos
Fil: Dai, Xiao-Qing. University of Alberta; Canadá
Fil: Chen, Xing-Zhen. University of Alberta; Canadá
Fil: Montalbetti, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
Fil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
Fil: Ausiello, Dennis A.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Cantiello, Horacio Fabio. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
Materia
VAP
PC2
cilia
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:11336/113747
Acceder
id CONICETDig_0761a3968b005ea2f75382a63cd53dce
oai_identifier_str oai:ri.conicet.gov.ar:11336/113747
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cellsRaychowdhury, Malay K.Ramos, Arnolt J.Zhang, PengMcLaughin, MargaretDai, Xiao-QingChen, Xing-ZhenMontalbetti, NicolasCantero, Maria del RocioAusiello, Dennis A.Cantiello, Horacio FabioVAPPC2ciliahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The primary cilium of renal epithelial cells is a nonmotile sensory organelle, implicated in mechanosensory transduction signals. Recent studies from our laboratory indicate that renal epithelial primary cilia display abundant channel activity; however, the presence and functional role of specific membrane receptors in this organelle are heretofore unknown. Here, we determined a functional signaling pathway associated with the type 2 vasopressin receptor (V2R) in primary cilia of renal epithelial cells. Besides their normal localization on basolateral membrane, V2R was expressed in primary cilia of LLC-PK1 renal epithelial cells. The presence of V2R in primary cilia was determined by spontaneous fluorescence of a V2R-gfp chimera and confirmed by immunocytochemical analysis of wild-type LLC-PK1 cells stained with anti-V2R antibodies and in LLC-PK1 cells overexpressing the V2R-Flag, with anti-Flag antibody. Ciliary V2R colocalized with adenylyl cyclase (AC) type V/VI in all cell types tested. Functional coupling of the receptors with AC was confirmed by measurement of cAMP production in isolated cilia and by testing AVP-induced cation-selective channel activity either in reconstituted lipid bilayers or subjected to membrane-attached patch clamping. Addition of either 10 μM AVP (trans) or forskolin (cis) in the presence but not the absence of ATP (1 mM, cis) stimulated cation-selective channel activity in ciliary membranes. This channel activity was reduced by addition of the PKA inhibitor PKI. The data provide the first demonstration for the presence of V2R in primary cilia of renal epithelial cells, and a functional cAMP-signaling pathway, which targets ciliary channel function and may help control the sensory function of the primary cilium.Fil: Raychowdhury, Malay K.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados UnidosFil: Ramos, Arnolt J.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados UnidosFil: Zhang, Peng. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados UnidosFil: McLaughin, Margaret. Harvard Medical School; Estados Unidos. Massachusetts General Hospital; Estados UnidosFil: Dai, Xiao-Qing. University of Alberta; CanadáFil: Chen, Xing-Zhen. University of Alberta; CanadáFil: Montalbetti, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaFil: Ausiello, Dennis A.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados UnidosFil: Cantiello, Horacio Fabio. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; ArgentinaAmerican Physiological Society2009-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/113747Raychowdhury, Malay K.; Ramos, Arnolt J.; Zhang, Peng; McLaughin, Margaret; Dai, Xiao-Qing; et al.; Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 296; 1; 1-2009; F87-F970363-61271931-857XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1152/ajprenal.90509.2008info:eu-repo/semantics/altIdentifier/url/https://journals.physiology.org/doi/full/10.1152/ajprenal.90509.2008info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:40:59Zoai:ri.conicet.gov.ar:11336/113747instacron: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:34982025-09-29 09:40:59.578CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
title Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
spellingShingle Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
Raychowdhury, Malay K.
VAP
PC2
cilia
title_short Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
title_full Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
title_fullStr Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
title_full_unstemmed Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
title_sort Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells
dc.creator.none.fl_str_mv Raychowdhury, Malay K.
Ramos, Arnolt J.
Zhang, Peng
McLaughin, Margaret
Dai, Xiao-Qing
Chen, Xing-Zhen
Montalbetti, Nicolas
Cantero, Maria del Rocio
Ausiello, Dennis A.
Cantiello, Horacio Fabio
author Raychowdhury, Malay K.
author_facet Raychowdhury, Malay K.
Ramos, Arnolt J.
Zhang, Peng
McLaughin, Margaret
Dai, Xiao-Qing
Chen, Xing-Zhen
Montalbetti, Nicolas
Cantero, Maria del Rocio
Ausiello, Dennis A.
Cantiello, Horacio Fabio
author_role author
author2 Ramos, Arnolt J.
Zhang, Peng
McLaughin, Margaret
Dai, Xiao-Qing
Chen, Xing-Zhen
Montalbetti, Nicolas
Cantero, Maria del Rocio
Ausiello, Dennis A.
Cantiello, Horacio Fabio
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv VAP
PC2
cilia
topic VAP
PC2
cilia
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The primary cilium of renal epithelial cells is a nonmotile sensory organelle, implicated in mechanosensory transduction signals. Recent studies from our laboratory indicate that renal epithelial primary cilia display abundant channel activity; however, the presence and functional role of specific membrane receptors in this organelle are heretofore unknown. Here, we determined a functional signaling pathway associated with the type 2 vasopressin receptor (V2R) in primary cilia of renal epithelial cells. Besides their normal localization on basolateral membrane, V2R was expressed in primary cilia of LLC-PK1 renal epithelial cells. The presence of V2R in primary cilia was determined by spontaneous fluorescence of a V2R-gfp chimera and confirmed by immunocytochemical analysis of wild-type LLC-PK1 cells stained with anti-V2R antibodies and in LLC-PK1 cells overexpressing the V2R-Flag, with anti-Flag antibody. Ciliary V2R colocalized with adenylyl cyclase (AC) type V/VI in all cell types tested. Functional coupling of the receptors with AC was confirmed by measurement of cAMP production in isolated cilia and by testing AVP-induced cation-selective channel activity either in reconstituted lipid bilayers or subjected to membrane-attached patch clamping. Addition of either 10 μM AVP (trans) or forskolin (cis) in the presence but not the absence of ATP (1 mM, cis) stimulated cation-selective channel activity in ciliary membranes. This channel activity was reduced by addition of the PKA inhibitor PKI. The data provide the first demonstration for the presence of V2R in primary cilia of renal epithelial cells, and a functional cAMP-signaling pathway, which targets ciliary channel function and may help control the sensory function of the primary cilium.
Fil: Raychowdhury, Malay K.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Ramos, Arnolt J.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Zhang, Peng. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: McLaughin, Margaret. Harvard Medical School; Estados Unidos. Massachusetts General Hospital; Estados Unidos
Fil: Dai, Xiao-Qing. University of Alberta; Canadá
Fil: Chen, Xing-Zhen. University of Alberta; Canadá
Fil: Montalbetti, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
Fil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
Fil: Ausiello, Dennis A.. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos
Fil: Cantiello, Horacio Fabio. Massachusetts General Hospital; Estados Unidos. Harvard Medical School; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Cardiológicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Cardiológicas; Argentina
description The primary cilium of renal epithelial cells is a nonmotile sensory organelle, implicated in mechanosensory transduction signals. Recent studies from our laboratory indicate that renal epithelial primary cilia display abundant channel activity; however, the presence and functional role of specific membrane receptors in this organelle are heretofore unknown. Here, we determined a functional signaling pathway associated with the type 2 vasopressin receptor (V2R) in primary cilia of renal epithelial cells. Besides their normal localization on basolateral membrane, V2R was expressed in primary cilia of LLC-PK1 renal epithelial cells. The presence of V2R in primary cilia was determined by spontaneous fluorescence of a V2R-gfp chimera and confirmed by immunocytochemical analysis of wild-type LLC-PK1 cells stained with anti-V2R antibodies and in LLC-PK1 cells overexpressing the V2R-Flag, with anti-Flag antibody. Ciliary V2R colocalized with adenylyl cyclase (AC) type V/VI in all cell types tested. Functional coupling of the receptors with AC was confirmed by measurement of cAMP production in isolated cilia and by testing AVP-induced cation-selective channel activity either in reconstituted lipid bilayers or subjected to membrane-attached patch clamping. Addition of either 10 μM AVP (trans) or forskolin (cis) in the presence but not the absence of ATP (1 mM, cis) stimulated cation-selective channel activity in ciliary membranes. This channel activity was reduced by addition of the PKA inhibitor PKI. The data provide the first demonstration for the presence of V2R in primary cilia of renal epithelial cells, and a functional cAMP-signaling pathway, which targets ciliary channel function and may help control the sensory function of the primary cilium.
publishDate 2009
dc.date.none.fl_str_mv 2009-01
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/11336/113747
Raychowdhury, Malay K.; Ramos, Arnolt J.; Zhang, Peng; McLaughin, Margaret; Dai, Xiao-Qing; et al.; Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 296; 1; 1-2009; F87-F97
0363-6127
1931-857X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/113747
identifier_str_mv Raychowdhury, Malay K.; Ramos, Arnolt J.; Zhang, Peng; McLaughin, Margaret; Dai, Xiao-Qing; et al.; Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 296; 1; 1-2009; F87-F97
0363-6127
1931-857X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1152/ajprenal.90509.2008
info:eu-repo/semantics/altIdentifier/url/https://journals.physiology.org/doi/full/10.1152/ajprenal.90509.2008
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
application/pdf
dc.publisher.none.fl_str_mv American Physiological Society
publisher.none.fl_str_mv American Physiological 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 13.070432

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