Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells

Autores
Torlaschi, Camila; Gutierrez Jofré, Gabriela Mabel; Rotstein, Nora Patricia; Simon, Maria Victoria
Año de publicación
2023
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Cell-cell interactions between retinal pigment epithelium (RPE) cells provide the retina with a physical and metabolic barrier, the disruption of which characterizes many inflammatory and prolifer- ative retinopathies. However, the underlying causes of this disrup- tion are still ill-defined. We showed that the bioactive sphingolipids sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) promote migration and inflammation in RPE cells. Using the human RPE cell line ARPE-19, we now analyzed whether S1P regulates cell morphology and RPE monolayer integrity. Inhibiting S1P synthe- sis with PF543, a sphingosine kinase 1 (SphK1) inhibitor, markedly decreased ARPE-19 cell migration in confluent cultures, without af- fecting cell survival. Using 50% confluent cultures, to better observe morphological changes, we determined that PF543 treatment pro- moted a remarkable cell retraction; highly elongated cells, absent in controls, augmented to 34±2% (p>0.01), their cell length/width ratio increasing to 5.3, from 1.6 in controls. S1P addition, 1 h af- ter PF543 treatment, restored cell morphology, reducing elongated cells to 8±1.4% (p>0.01), suggesting that S1P inside-out signaling is required for preserving cell morphology. In contrast, C1P addi- tion did not restore cell morphology in PF543-treated cells. When we preincubated cells with PF543 and JTE-013, a S1P2 receptor (S1P2) antagonist, before S1P addition, JTE-013 partially blocked S1P restoration of cell morphology. To analyze the mechanisms involved in cell adhesion, we determined distribution of paxillin, a scaffold protein in focal adhesions. While controls showed spot-like paxillin clusters in the cell periphery, these clusters disappeared in PF543-treated cells and were restored after S1P addition. These results suggest that inhibiting S1P synthesis leads to morphological changes and focal adhesion remodeling, and activation of the S1P/ S1P2 axis is required for preserving cell morphology and establish- ing focal adhesions.
Fil: Torlaschi, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Gutierrez Jofré, Gabriela Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Rotstein, Nora Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Simon, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
LXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Biología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Materia
SPHINGOLIPIDS
SPHINGOSINE-1-PHOSPHATE
RETINA PIGMENT EPITHELIA
FOCAL ADHESIONS
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:11336/229023
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cellsTorlaschi, CamilaGutierrez Jofré, Gabriela MabelRotstein, Nora PatriciaSimon, Maria VictoriaSPHINGOLIPIDSSPHINGOSINE-1-PHOSPHATERETINA PIGMENT EPITHELIAFOCAL ADHESIONShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Cell-cell interactions between retinal pigment epithelium (RPE) cells provide the retina with a physical and metabolic barrier, the disruption of which characterizes many inflammatory and prolifer- ative retinopathies. However, the underlying causes of this disrup- tion are still ill-defined. We showed that the bioactive sphingolipids sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) promote migration and inflammation in RPE cells. Using the human RPE cell line ARPE-19, we now analyzed whether S1P regulates cell morphology and RPE monolayer integrity. Inhibiting S1P synthe- sis with PF543, a sphingosine kinase 1 (SphK1) inhibitor, markedly decreased ARPE-19 cell migration in confluent cultures, without af- fecting cell survival. Using 50% confluent cultures, to better observe morphological changes, we determined that PF543 treatment pro- moted a remarkable cell retraction; highly elongated cells, absent in controls, augmented to 34±2% (p>0.01), their cell length/width ratio increasing to 5.3, from 1.6 in controls. S1P addition, 1 h af- ter PF543 treatment, restored cell morphology, reducing elongated cells to 8±1.4% (p>0.01), suggesting that S1P inside-out signaling is required for preserving cell morphology. In contrast, C1P addi- tion did not restore cell morphology in PF543-treated cells. When we preincubated cells with PF543 and JTE-013, a S1P2 receptor (S1P2) antagonist, before S1P addition, JTE-013 partially blocked S1P restoration of cell morphology. To analyze the mechanisms involved in cell adhesion, we determined distribution of paxillin, a scaffold protein in focal adhesions. While controls showed spot-like paxillin clusters in the cell periphery, these clusters disappeared in PF543-treated cells and were restored after S1P addition. These results suggest that inhibiting S1P synthesis leads to morphological changes and focal adhesion remodeling, and activation of the S1P/ S1P2 axis is required for preserving cell morphology and establish- ing focal adhesions.Fil: Torlaschi, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Gutierrez Jofré, Gabriela Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Rotstein, Nora Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Simon, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaLXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de BiologíaAsociación Argentina de Farmacología ExperimentalAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioFundación Revista Medicina2023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/229023Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells; LXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2023; 151-1510025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saic.org.ar/revista-medicinaNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:54:47Zoai:ri.conicet.gov.ar:11336/229023instacron: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 09:54:48.034CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
title Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
spellingShingle Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
Torlaschi, Camila
SPHINGOLIPIDS
SPHINGOSINE-1-PHOSPHATE
RETINA PIGMENT EPITHELIA
FOCAL ADHESIONS
title_short Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
title_full Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
title_fullStr Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
title_full_unstemmed Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
title_sort Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells
dc.creator.none.fl_str_mv Torlaschi, Camila
Gutierrez Jofré, Gabriela Mabel
Rotstein, Nora Patricia
Simon, Maria Victoria
author Torlaschi, Camila
author_facet Torlaschi, Camila
Gutierrez Jofré, Gabriela Mabel
Rotstein, Nora Patricia
Simon, Maria Victoria
author_role author
author2 Gutierrez Jofré, Gabriela Mabel
Rotstein, Nora Patricia
Simon, Maria Victoria
author2_role author
author
author
dc.subject.none.fl_str_mv SPHINGOLIPIDS
SPHINGOSINE-1-PHOSPHATE
RETINA PIGMENT EPITHELIA
FOCAL ADHESIONS
topic SPHINGOLIPIDS
SPHINGOSINE-1-PHOSPHATE
RETINA PIGMENT EPITHELIA
FOCAL ADHESIONS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Cell-cell interactions between retinal pigment epithelium (RPE) cells provide the retina with a physical and metabolic barrier, the disruption of which characterizes many inflammatory and prolifer- ative retinopathies. However, the underlying causes of this disrup- tion are still ill-defined. We showed that the bioactive sphingolipids sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) promote migration and inflammation in RPE cells. Using the human RPE cell line ARPE-19, we now analyzed whether S1P regulates cell morphology and RPE monolayer integrity. Inhibiting S1P synthe- sis with PF543, a sphingosine kinase 1 (SphK1) inhibitor, markedly decreased ARPE-19 cell migration in confluent cultures, without af- fecting cell survival. Using 50% confluent cultures, to better observe morphological changes, we determined that PF543 treatment pro- moted a remarkable cell retraction; highly elongated cells, absent in controls, augmented to 34±2% (p>0.01), their cell length/width ratio increasing to 5.3, from 1.6 in controls. S1P addition, 1 h af- ter PF543 treatment, restored cell morphology, reducing elongated cells to 8±1.4% (p>0.01), suggesting that S1P inside-out signaling is required for preserving cell morphology. In contrast, C1P addi- tion did not restore cell morphology in PF543-treated cells. When we preincubated cells with PF543 and JTE-013, a S1P2 receptor (S1P2) antagonist, before S1P addition, JTE-013 partially blocked S1P restoration of cell morphology. To analyze the mechanisms involved in cell adhesion, we determined distribution of paxillin, a scaffold protein in focal adhesions. While controls showed spot-like paxillin clusters in the cell periphery, these clusters disappeared in PF543-treated cells and were restored after S1P addition. These results suggest that inhibiting S1P synthesis leads to morphological changes and focal adhesion remodeling, and activation of the S1P/ S1P2 axis is required for preserving cell morphology and establish- ing focal adhesions.
Fil: Torlaschi, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Gutierrez Jofré, Gabriela Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Rotstein, Nora Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Simon, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
LXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Biología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
description Cell-cell interactions between retinal pigment epithelium (RPE) cells provide the retina with a physical and metabolic barrier, the disruption of which characterizes many inflammatory and prolifer- ative retinopathies. However, the underlying causes of this disrup- tion are still ill-defined. We showed that the bioactive sphingolipids sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) promote migration and inflammation in RPE cells. Using the human RPE cell line ARPE-19, we now analyzed whether S1P regulates cell morphology and RPE monolayer integrity. Inhibiting S1P synthe- sis with PF543, a sphingosine kinase 1 (SphK1) inhibitor, markedly decreased ARPE-19 cell migration in confluent cultures, without af- fecting cell survival. Using 50% confluent cultures, to better observe morphological changes, we determined that PF543 treatment pro- moted a remarkable cell retraction; highly elongated cells, absent in controls, augmented to 34±2% (p>0.01), their cell length/width ratio increasing to 5.3, from 1.6 in controls. S1P addition, 1 h af- ter PF543 treatment, restored cell morphology, reducing elongated cells to 8±1.4% (p>0.01), suggesting that S1P inside-out signaling is required for preserving cell morphology. In contrast, C1P addi- tion did not restore cell morphology in PF543-treated cells. When we preincubated cells with PF543 and JTE-013, a S1P2 receptor (S1P2) antagonist, before S1P addition, JTE-013 partially blocked S1P restoration of cell morphology. To analyze the mechanisms involved in cell adhesion, we determined distribution of paxillin, a scaffold protein in focal adhesions. While controls showed spot-like paxillin clusters in the cell periphery, these clusters disappeared in PF543-treated cells and were restored after S1P addition. These results suggest that inhibiting S1P synthesis leads to morphological changes and focal adhesion remodeling, and activation of the S1P/ S1P2 axis is required for preserving cell morphology and establish- ing focal adhesions.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
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info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/229023
Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells; LXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2023; 151-151
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/229023
identifier_str_mv Sphingosine-1-phosphate signaling is essential for preserving morphology and focal adhesions of retina pigment epithelial cells; LXVIII Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXV Jornadas Anuales de la Sociedad Argentina de Biología; LV Reunión Anual de la Asociación Argentina de Farmacología Experimental y VIII Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2023; 151-151
0025-7680
1669-9106
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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