Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors

Autores
Michelis, Germán Ariel; German, Olga Lorena; Rotstein, Nora Patricia; Politi, L.; Becerra, P.
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Small fragments from pigment epithelium-derived factor (PEDF), 44-mer and 17-mer, exhibit cytoprotective activity on photoreceptors and an R28 retinal progenitor cell line. They act by interacting with PEDF receptor (PEDF-R), a phospholipase encoded by the PNPLA2 gene. Recently, we found that PEDF prevents apoptosis and promotes apical localization of rhodopsin in retina photoreceptors in vitro. However, the effect of the peptides on primary retina cultures remains unknown. Here, we studied the effects of the 44-mer and 17-mer in developing retina neurons in culture. Pure neuronal cultures were prepared from retinas of rats at postnatal day 1. Cells expressed Pnpla2, and PEDF-R protein was detected at 1, 3 and 5 days after seeding. Immunolabeling with antibodies to PEDF-R and membrane marker Na/K ATPase, and wheat germ agglutinin revealed significant colocalization, implying PEDF-R distribution in retina cell surfaces. Rod photoreceptors and amacrine neurons were incubated in chemically defined medium supplemented at day 2 in culture with either PEDF synthetic peptide (10 nM); PEDF peptide plus PEDF-R P1 blocking peptide (100 nM) or PEDF-R inhibitor atglistatin; or vehicle (control) for 5 or 7 days in culture. Peptides 44-mer and 17-mer enhanced photoreceptor survival with time in culture relative to controls, as determined by TUNEL, Propidium Iodide and Annexin V/Mitotracker Flow Cytometry assays in vitro. The 44-mer and 17-mer peptide-mediated prevention of apoptosis involved the preservation of mitochondrial membrane potential. Peptide P1 pre-incubation or atglistatin abolished the survival effects of 44-mer and of 17-mer. Both peptides promoted a polarized rhodopsin localization resembling the natural development of the retina and PEDF effects. They also stimulated general axon outgrowth, especially in amacrine neurons, as axonal length doubled in these neurons in PEDF peptide-supplemented cultures, while a negative control peptide and presence of atglistatin or P1 did not. In addition to proving that PEDF fragments 44-mer and 17-mer are effective survival factors for retinal photoreceptors during development in vitro, the findings provide an insight for a mechanism of survival action in retinal neurons via interactions between the PEDF 17- mer and the PEDF-R P1 regions. PEDF peptides may play roles in neuronal differentiation, promoting the localization of opsin in apical processes and stimulating axonal outgrowth in amacrine neurons.
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. Public Healt Service; Estados Unidos
Fil: German, Olga Lorena. 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Politi, L.. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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, P.. Public Healt Service; Estados Unidos
The XXIII Biennial Meeting of the International Society for Eye Research
Irlanda
Reino Unido
International Society for Eye Research
Materia
PHOTORECEPTOR
PEDF
APOPTOSIS
DIFFERENTATION
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:11336/219989
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/219989
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina PhotoreceptorsMichelis, Germán ArielGerman, Olga LorenaRotstein, Nora PatriciaPoliti, L.Becerra, P.PHOTORECEPTORPEDFAPOPTOSISDIFFERENTATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Small fragments from pigment epithelium-derived factor (PEDF), 44-mer and 17-mer, exhibit cytoprotective activity on photoreceptors and an R28 retinal progenitor cell line. They act by interacting with PEDF receptor (PEDF-R), a phospholipase encoded by the PNPLA2 gene. Recently, we found that PEDF prevents apoptosis and promotes apical localization of rhodopsin in retina photoreceptors in vitro. However, the effect of the peptides on primary retina cultures remains unknown. Here, we studied the effects of the 44-mer and 17-mer in developing retina neurons in culture. Pure neuronal cultures were prepared from retinas of rats at postnatal day 1. Cells expressed Pnpla2, and PEDF-R protein was detected at 1, 3 and 5 days after seeding. Immunolabeling with antibodies to PEDF-R and membrane marker Na/K ATPase, and wheat germ agglutinin revealed significant colocalization, implying PEDF-R distribution in retina cell surfaces. Rod photoreceptors and amacrine neurons were incubated in chemically defined medium supplemented at day 2 in culture with either PEDF synthetic peptide (10 nM); PEDF peptide plus PEDF-R P1 blocking peptide (100 nM) or PEDF-R inhibitor atglistatin; or vehicle (control) for 5 or 7 days in culture. Peptides 44-mer and 17-mer enhanced photoreceptor survival with time in culture relative to controls, as determined by TUNEL, Propidium Iodide and Annexin V/Mitotracker Flow Cytometry assays in vitro. The 44-mer and 17-mer peptide-mediated prevention of apoptosis involved the preservation of mitochondrial membrane potential. Peptide P1 pre-incubation or atglistatin abolished the survival effects of 44-mer and of 17-mer. Both peptides promoted a polarized rhodopsin localization resembling the natural development of the retina and PEDF effects. They also stimulated general axon outgrowth, especially in amacrine neurons, as axonal length doubled in these neurons in PEDF peptide-supplemented cultures, while a negative control peptide and presence of atglistatin or P1 did not. In addition to proving that PEDF fragments 44-mer and 17-mer are effective survival factors for retinal photoreceptors during development in vitro, the findings provide an insight for a mechanism of survival action in retinal neurons via interactions between the PEDF 17- mer and the PEDF-R P1 regions. PEDF peptides may play roles in neuronal differentiation, promoting the localization of opsin in apical processes and stimulating axonal outgrowth in amacrine neurons.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. Public Healt Service; Estados UnidosFil: German, Olga Lorena. 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; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Politi, L.. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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, P.. Public Healt Service; Estados UnidosThe XXIII Biennial Meeting of the International Society for Eye ResearchIrlandaReino UnidoInternational Society for Eye ResearchInternational Society for Eye Research2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/219989Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors; The XXIII Biennial Meeting of the International Society for Eye Research; Irlanda; Reino Unido; 2018; 315-315CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.iser.org/meetings.htmlInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:07:13Zoai:ri.conicet.gov.ar:11336/219989instacron: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 10:07:13.447CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
title Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
spellingShingle Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
Michelis, Germán Ariel
PHOTORECEPTOR
PEDF
APOPTOSIS
DIFFERENTATION
title_short Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
title_full Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
title_fullStr Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
title_full_unstemmed Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
title_sort Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors
dc.creator.none.fl_str_mv Michelis, Germán Ariel
German, Olga Lorena
Rotstein, Nora Patricia
Politi, L.
Becerra, P.
author Michelis, Germán Ariel
author_facet Michelis, Germán Ariel
German, Olga Lorena
Rotstein, Nora Patricia
Politi, L.
Becerra, P.
author_role author
author2 German, Olga Lorena
Rotstein, Nora Patricia
Politi, L.
Becerra, P.
author2_role author
author
author
author
dc.subject.none.fl_str_mv PHOTORECEPTOR
PEDF
APOPTOSIS
DIFFERENTATION
topic PHOTORECEPTOR
PEDF
APOPTOSIS
DIFFERENTATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Small fragments from pigment epithelium-derived factor (PEDF), 44-mer and 17-mer, exhibit cytoprotective activity on photoreceptors and an R28 retinal progenitor cell line. They act by interacting with PEDF receptor (PEDF-R), a phospholipase encoded by the PNPLA2 gene. Recently, we found that PEDF prevents apoptosis and promotes apical localization of rhodopsin in retina photoreceptors in vitro. However, the effect of the peptides on primary retina cultures remains unknown. Here, we studied the effects of the 44-mer and 17-mer in developing retina neurons in culture. Pure neuronal cultures were prepared from retinas of rats at postnatal day 1. Cells expressed Pnpla2, and PEDF-R protein was detected at 1, 3 and 5 days after seeding. Immunolabeling with antibodies to PEDF-R and membrane marker Na/K ATPase, and wheat germ agglutinin revealed significant colocalization, implying PEDF-R distribution in retina cell surfaces. Rod photoreceptors and amacrine neurons were incubated in chemically defined medium supplemented at day 2 in culture with either PEDF synthetic peptide (10 nM); PEDF peptide plus PEDF-R P1 blocking peptide (100 nM) or PEDF-R inhibitor atglistatin; or vehicle (control) for 5 or 7 days in culture. Peptides 44-mer and 17-mer enhanced photoreceptor survival with time in culture relative to controls, as determined by TUNEL, Propidium Iodide and Annexin V/Mitotracker Flow Cytometry assays in vitro. The 44-mer and 17-mer peptide-mediated prevention of apoptosis involved the preservation of mitochondrial membrane potential. Peptide P1 pre-incubation or atglistatin abolished the survival effects of 44-mer and of 17-mer. Both peptides promoted a polarized rhodopsin localization resembling the natural development of the retina and PEDF effects. They also stimulated general axon outgrowth, especially in amacrine neurons, as axonal length doubled in these neurons in PEDF peptide-supplemented cultures, while a negative control peptide and presence of atglistatin or P1 did not. In addition to proving that PEDF fragments 44-mer and 17-mer are effective survival factors for retinal photoreceptors during development in vitro, the findings provide an insight for a mechanism of survival action in retinal neurons via interactions between the PEDF 17- mer and the PEDF-R P1 regions. PEDF peptides may play roles in neuronal differentiation, promoting the localization of opsin in apical processes and stimulating axonal outgrowth in amacrine neurons.
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. Public Healt Service; Estados Unidos
Fil: German, Olga Lorena. 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Politi, L.. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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, P.. Public Healt Service; Estados Unidos
The XXIII Biennial Meeting of the International Society for Eye Research
Irlanda
Reino Unido
International Society for Eye Research
description Small fragments from pigment epithelium-derived factor (PEDF), 44-mer and 17-mer, exhibit cytoprotective activity on photoreceptors and an R28 retinal progenitor cell line. They act by interacting with PEDF receptor (PEDF-R), a phospholipase encoded by the PNPLA2 gene. Recently, we found that PEDF prevents apoptosis and promotes apical localization of rhodopsin in retina photoreceptors in vitro. However, the effect of the peptides on primary retina cultures remains unknown. Here, we studied the effects of the 44-mer and 17-mer in developing retina neurons in culture. Pure neuronal cultures were prepared from retinas of rats at postnatal day 1. Cells expressed Pnpla2, and PEDF-R protein was detected at 1, 3 and 5 days after seeding. Immunolabeling with antibodies to PEDF-R and membrane marker Na/K ATPase, and wheat germ agglutinin revealed significant colocalization, implying PEDF-R distribution in retina cell surfaces. Rod photoreceptors and amacrine neurons were incubated in chemically defined medium supplemented at day 2 in culture with either PEDF synthetic peptide (10 nM); PEDF peptide plus PEDF-R P1 blocking peptide (100 nM) or PEDF-R inhibitor atglistatin; or vehicle (control) for 5 or 7 days in culture. Peptides 44-mer and 17-mer enhanced photoreceptor survival with time in culture relative to controls, as determined by TUNEL, Propidium Iodide and Annexin V/Mitotracker Flow Cytometry assays in vitro. The 44-mer and 17-mer peptide-mediated prevention of apoptosis involved the preservation of mitochondrial membrane potential. Peptide P1 pre-incubation or atglistatin abolished the survival effects of 44-mer and of 17-mer. Both peptides promoted a polarized rhodopsin localization resembling the natural development of the retina and PEDF effects. They also stimulated general axon outgrowth, especially in amacrine neurons, as axonal length doubled in these neurons in PEDF peptide-supplemented cultures, while a negative control peptide and presence of atglistatin or P1 did not. In addition to proving that PEDF fragments 44-mer and 17-mer are effective survival factors for retinal photoreceptors during development in vitro, the findings provide an insight for a mechanism of survival action in retinal neurons via interactions between the PEDF 17- mer and the PEDF-R P1 regions. PEDF peptides may play roles in neuronal differentiation, promoting the localization of opsin in apical processes and stimulating axonal outgrowth in amacrine neurons.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
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/11336/219989
Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors; The XXIII Biennial Meeting of the International Society for Eye Research; Irlanda; Reino Unido; 2018; 315-315
CONICET Digital
CONICET
url http://hdl.handle.net/11336/219989
identifier_str_mv Peptides Derived from PEDF Prevent Apoptosis and Promote Apical Localization of Rhodopsin in Retina Photoreceptors; The XXIII Biennial Meeting of the International Society for Eye Research; Irlanda; Reino Unido; 2018; 315-315
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.iser.org/meetings.html
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 International Society for Eye Research
publisher.none.fl_str_mv International Society for Eye Research
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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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