Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons

Autores
Politi, Luis Enrique; Michelis, Germán Ariel; Becerra, Patricia; Rotstein, Nora Patricia
Año de publicación
2017
Idioma
español castellano
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
PEDF promotes survival of photoreceptor cells and in an R28 retinal progenitor cell line by binding to PEDF membrane receptors. However, the intracellular molecular effects of PEDF in retina neurons are still unknown. Here we investigated these effects using pure neuronal cultures of photoreceptors and amacrine neurons, prepared from newborn rat retina and grown in a chemically defined medium. Cultures were supplemented at day 2 with either recombinant human PEDF (10 nM); PEDF plus P1 (a PEDF receptor blocking peptide, 100nM); or vehicle (control). At day 5, cells were fixed and cell death and apoptosis were evaluated by DAPI staining, Propidium Iodide and TUNEL assays, respectively. Mitochondrial activity was assessed with Mitotracker and opsin expression and axonal outgrowth were determined by immunocytochemical methods. PEDF decreased the percentages of Propidium Iodide and TUNEL-positive photoreceptors by 55% and 65%, respectively, with respect to controls without PEDF, and significantly prevented the loss of mitochondrial membrane potential. Pre-incubation with the blocking P1 peptide abolished PEDF effects. PEDF had no effect on opsin expression but promoted its apical localization in photoreceptors as occurs in mature retinas, in contrast to its diffuse distribution on the cell body in controls. PEDF also stimulated axon outgrowth in amacrine neurons, by 2-fold, in neurons treated with PEDF, compared to controls. These effects were blocked by the blocking P1 peptide pre-treatment. In summary, this work shows that PEDF acts as a survival factor for photoreceptors, preserving their mitochondrial functionality, and induces the differentiation of amacrine and photoreceptor neurons during their development in vitro.
Fil: Politi, Luis Enrique. 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: Michelis, Germán Ariel. 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: Becerra, Patricia. No especifíca;
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
LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina De Investigación Clínica
Sociedad Argentina De Investigación Bioquímica Y Biología Molecular
Sociedad Argentina De Inmunología
Sociedad Argentina De Andrología
Sociedad Argentina De Biofísica
Sociedad Argentina De Biología
Sociedad Argentina De Farmacología Experimental
Sociedad Argentina De Fisiología
Sociedad Argentina De Hematología
Sociedad Argentina De Protozoología
Materia
PEDF
apoptosis prevention
retina photoreceptors
amacrine neurons
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:11336/244017
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/244017
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neuronsPoliti, Luis EnriqueMichelis, Germán ArielBecerra, PatriciaRotstein, Nora PatriciaPEDFapoptosis preventionretina photoreceptorsamacrine neuronshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1PEDF promotes survival of photoreceptor cells and in an R28 retinal progenitor cell line by binding to PEDF membrane receptors. However, the intracellular molecular effects of PEDF in retina neurons are still unknown. Here we investigated these effects using pure neuronal cultures of photoreceptors and amacrine neurons, prepared from newborn rat retina and grown in a chemically defined medium. Cultures were supplemented at day 2 with either recombinant human PEDF (10 nM); PEDF plus P1 (a PEDF receptor blocking peptide, 100nM); or vehicle (control). At day 5, cells were fixed and cell death and apoptosis were evaluated by DAPI staining, Propidium Iodide and TUNEL assays, respectively. Mitochondrial activity was assessed with Mitotracker and opsin expression and axonal outgrowth were determined by immunocytochemical methods. PEDF decreased the percentages of Propidium Iodide and TUNEL-positive photoreceptors by 55% and 65%, respectively, with respect to controls without PEDF, and significantly prevented the loss of mitochondrial membrane potential. Pre-incubation with the blocking P1 peptide abolished PEDF effects. PEDF had no effect on opsin expression but promoted its apical localization in photoreceptors as occurs in mature retinas, in contrast to its diffuse distribution on the cell body in controls. PEDF also stimulated axon outgrowth in amacrine neurons, by 2-fold, in neurons treated with PEDF, compared to controls. These effects were blocked by the blocking P1 peptide pre-treatment. In summary, this work shows that PEDF acts as a survival factor for photoreceptors, preserving their mitochondrial functionality, and induces the differentiation of amacrine and photoreceptor neurons during their development in vitro.Fil: Politi, Luis Enrique. 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: Michelis, Germán Ariel. 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: Becerra, Patricia. No especifíca;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; ArgentinaLXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de ProtozoologíaBuenos AiresArgentinaSociedad Argentina De Investigación ClínicaSociedad Argentina De Investigación Bioquímica Y Biología MolecularSociedad Argentina De InmunologíaSociedad Argentina De AndrologíaSociedad Argentina De BiofísicaSociedad Argentina De BiologíaSociedad Argentina De Farmacología ExperimentalSociedad Argentina De FisiologíaSociedad Argentina De HematologíaSociedad Argentina De ProtozoologíaFundación Revista Medicina2017info: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/11336/244017Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 222-2220076-60461669-9106CONICET DigitalCONICETspainfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:14Zoai:ri.conicet.gov.ar:11336/244017instacron: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 10:02:14.615CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
title Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
spellingShingle Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
Politi, Luis Enrique
PEDF
apoptosis prevention
retina photoreceptors
amacrine neurons
title_short Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
title_full Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
title_fullStr Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
title_full_unstemmed Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
title_sort Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons
dc.creator.none.fl_str_mv Politi, Luis Enrique
Michelis, Germán Ariel
Becerra, Patricia
Rotstein, Nora Patricia
author Politi, Luis Enrique
author_facet Politi, Luis Enrique
Michelis, Germán Ariel
Becerra, Patricia
Rotstein, Nora Patricia
author_role author
author2 Michelis, Germán Ariel
Becerra, Patricia
Rotstein, Nora Patricia
author2_role author
author
author
dc.subject.none.fl_str_mv PEDF
apoptosis prevention
retina photoreceptors
amacrine neurons
topic PEDF
apoptosis prevention
retina photoreceptors
amacrine neurons
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv PEDF promotes survival of photoreceptor cells and in an R28 retinal progenitor cell line by binding to PEDF membrane receptors. However, the intracellular molecular effects of PEDF in retina neurons are still unknown. Here we investigated these effects using pure neuronal cultures of photoreceptors and amacrine neurons, prepared from newborn rat retina and grown in a chemically defined medium. Cultures were supplemented at day 2 with either recombinant human PEDF (10 nM); PEDF plus P1 (a PEDF receptor blocking peptide, 100nM); or vehicle (control). At day 5, cells were fixed and cell death and apoptosis were evaluated by DAPI staining, Propidium Iodide and TUNEL assays, respectively. Mitochondrial activity was assessed with Mitotracker and opsin expression and axonal outgrowth were determined by immunocytochemical methods. PEDF decreased the percentages of Propidium Iodide and TUNEL-positive photoreceptors by 55% and 65%, respectively, with respect to controls without PEDF, and significantly prevented the loss of mitochondrial membrane potential. Pre-incubation with the blocking P1 peptide abolished PEDF effects. PEDF had no effect on opsin expression but promoted its apical localization in photoreceptors as occurs in mature retinas, in contrast to its diffuse distribution on the cell body in controls. PEDF also stimulated axon outgrowth in amacrine neurons, by 2-fold, in neurons treated with PEDF, compared to controls. These effects were blocked by the blocking P1 peptide pre-treatment. In summary, this work shows that PEDF acts as a survival factor for photoreceptors, preserving their mitochondrial functionality, and induces the differentiation of amacrine and photoreceptor neurons during their development in vitro.
Fil: Politi, Luis Enrique. 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: Michelis, Germán Ariel. 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: Becerra, Patricia. No especifíca;
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
LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina De Investigación Clínica
Sociedad Argentina De Investigación Bioquímica Y Biología Molecular
Sociedad Argentina De Inmunología
Sociedad Argentina De Andrología
Sociedad Argentina De Biofísica
Sociedad Argentina De Biología
Sociedad Argentina De Farmacología Experimental
Sociedad Argentina De Fisiología
Sociedad Argentina De Hematología
Sociedad Argentina De Protozoología
description PEDF promotes survival of photoreceptor cells and in an R28 retinal progenitor cell line by binding to PEDF membrane receptors. However, the intracellular molecular effects of PEDF in retina neurons are still unknown. Here we investigated these effects using pure neuronal cultures of photoreceptors and amacrine neurons, prepared from newborn rat retina and grown in a chemically defined medium. Cultures were supplemented at day 2 with either recombinant human PEDF (10 nM); PEDF plus P1 (a PEDF receptor blocking peptide, 100nM); or vehicle (control). At day 5, cells were fixed and cell death and apoptosis were evaluated by DAPI staining, Propidium Iodide and TUNEL assays, respectively. Mitochondrial activity was assessed with Mitotracker and opsin expression and axonal outgrowth were determined by immunocytochemical methods. PEDF decreased the percentages of Propidium Iodide and TUNEL-positive photoreceptors by 55% and 65%, respectively, with respect to controls without PEDF, and significantly prevented the loss of mitochondrial membrane potential. Pre-incubation with the blocking P1 peptide abolished PEDF effects. PEDF had no effect on opsin expression but promoted its apical localization in photoreceptors as occurs in mature retinas, in contrast to its diffuse distribution on the cell body in controls. PEDF also stimulated axon outgrowth in amacrine neurons, by 2-fold, in neurons treated with PEDF, compared to controls. These effects were blocked by the blocking P1 peptide pre-treatment. In summary, this work shows that PEDF acts as a survival factor for photoreceptors, preserving their mitochondrial functionality, and induces the differentiation of amacrine and photoreceptor neurons during their development in vitro.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/244017
Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 222-222
0076-6046
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244017
identifier_str_mv Survival and differentiation effects of pigment epithelium derived factor (pedf) on retina neurons; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 222-222
0076-6046
1669-9106
CONICET Digital
CONICET
dc.language.none.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdf
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.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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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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