Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells
- Autores
- Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio
- Año de publicación
- 2022
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Polycystin-2 (PC2, TRPP2) is a Ca2+ permeable nonselective cation channel whose dysfunction generates autosomal dominant polycystic kidney disease (ADPKD). PC2 is present in different cell locations, including the primary cilium of renal epithelial cells. However, little is known as to whether PC2 contributes to the primary cilium structure. Here, we explored the effect(s) of external Ca2+, PC2 channel blockers, and PKD2 gene silencing on the length of primary cilia in wild-type LLC-PK1 renal epithelial cells. Confluent cell monolayers were fixed and immuno-labeled with an anti-acetylated α-tubulin antibody to identify primary cilia and measure their length. Although primary cilia length measurements did not follow a Normal distribution, the data were normalized by Box-Cox transformation rendering statistical differences under all experimental conditions. Cells exposed to high external Ca2+ (6.2 mM) decreased a 13.5% (p < 0.001) primary cilia length as compared to controls (1.2 mM Ca2+). In contrast, the PC2 inhibitors amiloride (200 μM) and LiCl (10 mM), both increased primary ciliary length by 33.2% (p < 0.001), and 17.4% (p < 0.001), respectively. PKD2 gene silencing by siRNA elicited a statistically significant, 10.3% (p < 0.001) increase in primary cilia length compared to their respective scrambled RNA transfected cells. The data indicate that conditions that regulate PC2 function or gene expression modify the length of primary cilia in renal epithelial cells. Blocking of PC2 mitigates the effects of elevated external Ca2+ concentration on primary cilia length. Proper regulation of PC2 function in the primary cilium may be essential in the onset of mechanisms that trigger cyst formation in ADPKD.
Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina
Fil: Pérez, Paula Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina
Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina
Fil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina - Materia
-
ADPKD
CALCIUM
LITHIUM
POLYCYSTIN-2
PRIMARY CILIA - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/213490
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Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cellsScarinci, María NoeliaPérez, Paula LucianaCantiello, Horacio FabioCantero, Maria del RocioADPKDCALCIUMLITHIUMPOLYCYSTIN-2PRIMARY CILIAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Polycystin-2 (PC2, TRPP2) is a Ca2+ permeable nonselective cation channel whose dysfunction generates autosomal dominant polycystic kidney disease (ADPKD). PC2 is present in different cell locations, including the primary cilium of renal epithelial cells. However, little is known as to whether PC2 contributes to the primary cilium structure. Here, we explored the effect(s) of external Ca2+, PC2 channel blockers, and PKD2 gene silencing on the length of primary cilia in wild-type LLC-PK1 renal epithelial cells. Confluent cell monolayers were fixed and immuno-labeled with an anti-acetylated α-tubulin antibody to identify primary cilia and measure their length. Although primary cilia length measurements did not follow a Normal distribution, the data were normalized by Box-Cox transformation rendering statistical differences under all experimental conditions. Cells exposed to high external Ca2+ (6.2 mM) decreased a 13.5% (p < 0.001) primary cilia length as compared to controls (1.2 mM Ca2+). In contrast, the PC2 inhibitors amiloride (200 μM) and LiCl (10 mM), both increased primary ciliary length by 33.2% (p < 0.001), and 17.4% (p < 0.001), respectively. PKD2 gene silencing by siRNA elicited a statistically significant, 10.3% (p < 0.001) increase in primary cilia length compared to their respective scrambled RNA transfected cells. The data indicate that conditions that regulate PC2 function or gene expression modify the length of primary cilia in renal epithelial cells. Blocking of PC2 mitigates the effects of elevated external Ca2+ concentration on primary cilia length. Proper regulation of PC2 function in the primary cilium may be essential in the onset of mechanisms that trigger cyst formation in ADPKD.Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; ArgentinaFil: Pérez, Paula Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; ArgentinaFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; ArgentinaFil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; ArgentinaFrontiers Media2022-09info: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/11336/213490Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells; Frontiers Media; Frontiers in Physiology; 13; 9-2022; 1-131664-042XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fphys.2022.995473info:eu-repo/semantics/altIdentifier/doi/10.3389/fphys.2022.995473info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:30:27Zoai:ri.conicet.gov.ar:11336/213490instacron: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:34982025-10-15 15:30:28.121CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
title |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
spellingShingle |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells Scarinci, María Noelia ADPKD CALCIUM LITHIUM POLYCYSTIN-2 PRIMARY CILIA |
title_short |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
title_full |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
title_fullStr |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
title_full_unstemmed |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
title_sort |
Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells |
dc.creator.none.fl_str_mv |
Scarinci, María Noelia Pérez, Paula Luciana Cantiello, Horacio Fabio Cantero, Maria del Rocio |
author |
Scarinci, María Noelia |
author_facet |
Scarinci, María Noelia Pérez, Paula Luciana Cantiello, Horacio Fabio Cantero, Maria del Rocio |
author_role |
author |
author2 |
Pérez, Paula Luciana Cantiello, Horacio Fabio Cantero, Maria del Rocio |
author2_role |
author author author |
dc.subject.none.fl_str_mv |
ADPKD CALCIUM LITHIUM POLYCYSTIN-2 PRIMARY CILIA |
topic |
ADPKD CALCIUM LITHIUM POLYCYSTIN-2 PRIMARY 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 |
Polycystin-2 (PC2, TRPP2) is a Ca2+ permeable nonselective cation channel whose dysfunction generates autosomal dominant polycystic kidney disease (ADPKD). PC2 is present in different cell locations, including the primary cilium of renal epithelial cells. However, little is known as to whether PC2 contributes to the primary cilium structure. Here, we explored the effect(s) of external Ca2+, PC2 channel blockers, and PKD2 gene silencing on the length of primary cilia in wild-type LLC-PK1 renal epithelial cells. Confluent cell monolayers were fixed and immuno-labeled with an anti-acetylated α-tubulin antibody to identify primary cilia and measure their length. Although primary cilia length measurements did not follow a Normal distribution, the data were normalized by Box-Cox transformation rendering statistical differences under all experimental conditions. Cells exposed to high external Ca2+ (6.2 mM) decreased a 13.5% (p < 0.001) primary cilia length as compared to controls (1.2 mM Ca2+). In contrast, the PC2 inhibitors amiloride (200 μM) and LiCl (10 mM), both increased primary ciliary length by 33.2% (p < 0.001), and 17.4% (p < 0.001), respectively. PKD2 gene silencing by siRNA elicited a statistically significant, 10.3% (p < 0.001) increase in primary cilia length compared to their respective scrambled RNA transfected cells. The data indicate that conditions that regulate PC2 function or gene expression modify the length of primary cilia in renal epithelial cells. Blocking of PC2 mitigates the effects of elevated external Ca2+ concentration on primary cilia length. Proper regulation of PC2 function in the primary cilium may be essential in the onset of mechanisms that trigger cyst formation in ADPKD. Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Pérez, Paula Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina |
description |
Polycystin-2 (PC2, TRPP2) is a Ca2+ permeable nonselective cation channel whose dysfunction generates autosomal dominant polycystic kidney disease (ADPKD). PC2 is present in different cell locations, including the primary cilium of renal epithelial cells. However, little is known as to whether PC2 contributes to the primary cilium structure. Here, we explored the effect(s) of external Ca2+, PC2 channel blockers, and PKD2 gene silencing on the length of primary cilia in wild-type LLC-PK1 renal epithelial cells. Confluent cell monolayers were fixed and immuno-labeled with an anti-acetylated α-tubulin antibody to identify primary cilia and measure their length. Although primary cilia length measurements did not follow a Normal distribution, the data were normalized by Box-Cox transformation rendering statistical differences under all experimental conditions. Cells exposed to high external Ca2+ (6.2 mM) decreased a 13.5% (p < 0.001) primary cilia length as compared to controls (1.2 mM Ca2+). In contrast, the PC2 inhibitors amiloride (200 μM) and LiCl (10 mM), both increased primary ciliary length by 33.2% (p < 0.001), and 17.4% (p < 0.001), respectively. PKD2 gene silencing by siRNA elicited a statistically significant, 10.3% (p < 0.001) increase in primary cilia length compared to their respective scrambled RNA transfected cells. The data indicate that conditions that regulate PC2 function or gene expression modify the length of primary cilia in renal epithelial cells. Blocking of PC2 mitigates the effects of elevated external Ca2+ concentration on primary cilia length. Proper regulation of PC2 function in the primary cilium may be essential in the onset of mechanisms that trigger cyst formation in ADPKD. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-09 |
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/11336/213490 Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells; Frontiers Media; Frontiers in Physiology; 13; 9-2022; 1-13 1664-042X CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/213490 |
identifier_str_mv |
Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells; Frontiers Media; Frontiers in Physiology; 13; 9-2022; 1-13 1664-042X 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://www.frontiersin.org/articles/10.3389/fphys.2022.995473 info:eu-repo/semantics/altIdentifier/doi/10.3389/fphys.2022.995473 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Frontiers Media |
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Frontiers Media |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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13.22299 |