ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa

Autores
Vallese, Harmonie Agostina; Viera, Solange; Colo, Georgina Pamela; German, Lorena
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Purpose : In Retinitis Pigmentosa the progressive loss of photoreceptors (PRs) leads to vision impairment. Our previous work showed that Müller glial cells (MGCs), specialized retinal cells that support PRs survival, are affected in the retinal degeneration mouse model (rd1). Even before PRs degeneration, rd1 MGCs evidence a reduced regenerative potential, incapacity to form lamellipodia and impairment of their neuroprotective role on PRs. Whether the extracellular matrix (ECM) is affected in rd1 retinas, altering MGCs adhesion and functionality and contributing to PRs death remains unclear. The aim of this study was to investigate ECM protein expression in rd1 retinas prior to the peak of PRs degeneration and assess whether ECM pretreatment can restore the morphology and functionality of rd1 MGCs in vitro. Methods : Mixed neuron-glial cultures were prepared from postnatal day 2 rd1 and wt mouse retinas. We analyzed SPARC and Fibronectin expression (ECM proteins), Paxillin (focal adhesions, FAs), and actin by immunocytochemistry, and measured the neuron-glial cluster area with Image J software. We obtained ECM (SPARC)-enriched conditioned media (CM) from retinoblastoma RN22 cell line cultures. We then grew rd1 cultures on dishes pre-treated or not with ECM-CM to evaluate its effects on neuron-glial cluster area, FAs, actin cytoskeleton, cell proliferation, measured through BrdU nucleotide incorporation, and cell death, assessed by DAPI staining. Results : Neuron-glial clusters were smaller, whereas expression of SPARC was significantly lower (p=0.0059) and that of fibrillary fibronectin higher in rd1 than in wt neuron-glial cultures (p=0.0002). Rd1 MGCs had shorter FA (p= 0.0059), which showed mainly a cortical distribution (p=0.001), contrary to wt MGCs. Actin displayed a cortical distribution, and higher fluorescence intensity (p= 0.004) in rd1 compared to wt cultures. Noteworthy, dish pre-treatment with ECM-CM increased the size of neuron-glial clusters (p= 0.0006), restored the distribution of glial FAs, and reduced phalloidin fluorescence intensity (p= 0.0005). Moreover, ECM-CM pre-treatment also stimulated MGCs proliferation (p= 0.021) and increased PRs survival (p= 0.0003) in rd1 cultures. Conclusions : Our results suggest that expression of ECM proteins and FAs assembly are altered in rd1 MGCs and enrichment with ECM restores MGCs functionality, contributing to PRs survival
Fil: Vallese, Harmonie Agostina. 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: Viera, Solange. 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: Colo, Georgina Pamela. 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: German, 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
ARVO Annual Meeting
Seattle
Estados Unidos
Association for Research in Vision and Ophthalmology
Materia
CGM
Matriz Extracelullar
Fotorreceptores
Neurodegeneración
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:11336/243996
Acceder
id CONICETDig_3d010226d4869f45853669b1e3293285
oai_identifier_str oai:ri.conicet.gov.ar:11336/243996
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis PigmentosaVallese, Harmonie AgostinaViera, SolangeColo, Georgina PamelaGerman, LorenaCGMMatriz ExtracelullarFotorreceptoresNeurodegeneraciónhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Purpose : In Retinitis Pigmentosa the progressive loss of photoreceptors (PRs) leads to vision impairment. Our previous work showed that Müller glial cells (MGCs), specialized retinal cells that support PRs survival, are affected in the retinal degeneration mouse model (rd1). Even before PRs degeneration, rd1 MGCs evidence a reduced regenerative potential, incapacity to form lamellipodia and impairment of their neuroprotective role on PRs. Whether the extracellular matrix (ECM) is affected in rd1 retinas, altering MGCs adhesion and functionality and contributing to PRs death remains unclear. The aim of this study was to investigate ECM protein expression in rd1 retinas prior to the peak of PRs degeneration and assess whether ECM pretreatment can restore the morphology and functionality of rd1 MGCs in vitro. Methods : Mixed neuron-glial cultures were prepared from postnatal day 2 rd1 and wt mouse retinas. We analyzed SPARC and Fibronectin expression (ECM proteins), Paxillin (focal adhesions, FAs), and actin by immunocytochemistry, and measured the neuron-glial cluster area with Image J software. We obtained ECM (SPARC)-enriched conditioned media (CM) from retinoblastoma RN22 cell line cultures. We then grew rd1 cultures on dishes pre-treated or not with ECM-CM to evaluate its effects on neuron-glial cluster area, FAs, actin cytoskeleton, cell proliferation, measured through BrdU nucleotide incorporation, and cell death, assessed by DAPI staining. Results : Neuron-glial clusters were smaller, whereas expression of SPARC was significantly lower (p=0.0059) and that of fibrillary fibronectin higher in rd1 than in wt neuron-glial cultures (p=0.0002). Rd1 MGCs had shorter FA (p= 0.0059), which showed mainly a cortical distribution (p=0.001), contrary to wt MGCs. Actin displayed a cortical distribution, and higher fluorescence intensity (p= 0.004) in rd1 compared to wt cultures. Noteworthy, dish pre-treatment with ECM-CM increased the size of neuron-glial clusters (p= 0.0006), restored the distribution of glial FAs, and reduced phalloidin fluorescence intensity (p= 0.0005). Moreover, ECM-CM pre-treatment also stimulated MGCs proliferation (p= 0.021) and increased PRs survival (p= 0.0003) in rd1 cultures. Conclusions : Our results suggest that expression of ECM proteins and FAs assembly are altered in rd1 MGCs and enrichment with ECM restores MGCs functionality, contributing to PRs survivalFil: Vallese, Harmonie Agostina. 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: Viera, Solange. 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: Colo, Georgina Pamela. 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: German, 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; ArgentinaARVO Annual MeetingSeattleEstados UnidosAssociation for Research in Vision and OphthalmologyAssociation for Research in Vision and Ophthalmology.2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/243996ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa; ARVO Annual Meeting; Seattle; Estados Unidos; 2024; 1-11552-5783CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://iovs.arvojournals.org/article.aspx?articleid=2796669Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:43:46Zoai:ri.conicet.gov.ar:11336/243996instacron: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 09:43:46.536CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
title ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
spellingShingle ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
Vallese, Harmonie Agostina
CGM
Matriz Extracelullar
Fotorreceptores
Neurodegeneración
title_short ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
title_full ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
title_fullStr ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
title_full_unstemmed ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
title_sort ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa
dc.creator.none.fl_str_mv Vallese, Harmonie Agostina
Viera, Solange
Colo, Georgina Pamela
German, Lorena
author Vallese, Harmonie Agostina
author_facet Vallese, Harmonie Agostina
Viera, Solange
Colo, Georgina Pamela
German, Lorena
author_role author
author2 Viera, Solange
Colo, Georgina Pamela
German, Lorena
author2_role author
author
author
dc.subject.none.fl_str_mv CGM
Matriz Extracelullar
Fotorreceptores
Neurodegeneración
topic CGM
Matriz Extracelullar
Fotorreceptores
Neurodegeneración
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Purpose : In Retinitis Pigmentosa the progressive loss of photoreceptors (PRs) leads to vision impairment. Our previous work showed that Müller glial cells (MGCs), specialized retinal cells that support PRs survival, are affected in the retinal degeneration mouse model (rd1). Even before PRs degeneration, rd1 MGCs evidence a reduced regenerative potential, incapacity to form lamellipodia and impairment of their neuroprotective role on PRs. Whether the extracellular matrix (ECM) is affected in rd1 retinas, altering MGCs adhesion and functionality and contributing to PRs death remains unclear. The aim of this study was to investigate ECM protein expression in rd1 retinas prior to the peak of PRs degeneration and assess whether ECM pretreatment can restore the morphology and functionality of rd1 MGCs in vitro. Methods : Mixed neuron-glial cultures were prepared from postnatal day 2 rd1 and wt mouse retinas. We analyzed SPARC and Fibronectin expression (ECM proteins), Paxillin (focal adhesions, FAs), and actin by immunocytochemistry, and measured the neuron-glial cluster area with Image J software. We obtained ECM (SPARC)-enriched conditioned media (CM) from retinoblastoma RN22 cell line cultures. We then grew rd1 cultures on dishes pre-treated or not with ECM-CM to evaluate its effects on neuron-glial cluster area, FAs, actin cytoskeleton, cell proliferation, measured through BrdU nucleotide incorporation, and cell death, assessed by DAPI staining. Results : Neuron-glial clusters were smaller, whereas expression of SPARC was significantly lower (p=0.0059) and that of fibrillary fibronectin higher in rd1 than in wt neuron-glial cultures (p=0.0002). Rd1 MGCs had shorter FA (p= 0.0059), which showed mainly a cortical distribution (p=0.001), contrary to wt MGCs. Actin displayed a cortical distribution, and higher fluorescence intensity (p= 0.004) in rd1 compared to wt cultures. Noteworthy, dish pre-treatment with ECM-CM increased the size of neuron-glial clusters (p= 0.0006), restored the distribution of glial FAs, and reduced phalloidin fluorescence intensity (p= 0.0005). Moreover, ECM-CM pre-treatment also stimulated MGCs proliferation (p= 0.021) and increased PRs survival (p= 0.0003) in rd1 cultures. Conclusions : Our results suggest that expression of ECM proteins and FAs assembly are altered in rd1 MGCs and enrichment with ECM restores MGCs functionality, contributing to PRs survival
Fil: Vallese, Harmonie Agostina. 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: Viera, Solange. 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: Colo, Georgina Pamela. 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: German, 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
ARVO Annual Meeting
Seattle
Estados Unidos
Association for Research in Vision and Ophthalmology
description Purpose : In Retinitis Pigmentosa the progressive loss of photoreceptors (PRs) leads to vision impairment. Our previous work showed that Müller glial cells (MGCs), specialized retinal cells that support PRs survival, are affected in the retinal degeneration mouse model (rd1). Even before PRs degeneration, rd1 MGCs evidence a reduced regenerative potential, incapacity to form lamellipodia and impairment of their neuroprotective role on PRs. Whether the extracellular matrix (ECM) is affected in rd1 retinas, altering MGCs adhesion and functionality and contributing to PRs death remains unclear. The aim of this study was to investigate ECM protein expression in rd1 retinas prior to the peak of PRs degeneration and assess whether ECM pretreatment can restore the morphology and functionality of rd1 MGCs in vitro. Methods : Mixed neuron-glial cultures were prepared from postnatal day 2 rd1 and wt mouse retinas. We analyzed SPARC and Fibronectin expression (ECM proteins), Paxillin (focal adhesions, FAs), and actin by immunocytochemistry, and measured the neuron-glial cluster area with Image J software. We obtained ECM (SPARC)-enriched conditioned media (CM) from retinoblastoma RN22 cell line cultures. We then grew rd1 cultures on dishes pre-treated or not with ECM-CM to evaluate its effects on neuron-glial cluster area, FAs, actin cytoskeleton, cell proliferation, measured through BrdU nucleotide incorporation, and cell death, assessed by DAPI staining. Results : Neuron-glial clusters were smaller, whereas expression of SPARC was significantly lower (p=0.0059) and that of fibrillary fibronectin higher in rd1 than in wt neuron-glial cultures (p=0.0002). Rd1 MGCs had shorter FA (p= 0.0059), which showed mainly a cortical distribution (p=0.001), contrary to wt MGCs. Actin displayed a cortical distribution, and higher fluorescence intensity (p= 0.004) in rd1 compared to wt cultures. Noteworthy, dish pre-treatment with ECM-CM increased the size of neuron-glial clusters (p= 0.0006), restored the distribution of glial FAs, and reduced phalloidin fluorescence intensity (p= 0.0005). Moreover, ECM-CM pre-treatment also stimulated MGCs proliferation (p= 0.021) and increased PRs survival (p= 0.0003) in rd1 cultures. Conclusions : Our results suggest that expression of ECM proteins and FAs assembly are altered in rd1 MGCs and enrichment with ECM restores MGCs functionality, contributing to PRs survival
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
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/11336/243996
ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa; ARVO Annual Meeting; Seattle; Estados Unidos; 2024; 1-1
1552-5783
CONICET Digital
CONICET
url http://hdl.handle.net/11336/243996
identifier_str_mv ECM enrichment restores the morphology and functionality of Müller Glial Cells in the rd1 Model of Retinitis Pigmentosa; ARVO Annual Meeting; Seattle; Estados Unidos; 2024; 1-1
1552-5783
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://iovs.arvojournals.org/article.aspx?articleid=2796669
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 Association for Research in Vision and Ophthalmology.
publisher.none.fl_str_mv Association for Research in Vision and Ophthalmology.
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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