IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility
- Autores
- Lorenc, Valeria Erika; Jaldín Fincati, Javier Roberto; Luna Pinto, Jose Domingo; Chiabrando, Gustavo Alberto; Sanchez, Maria Cecilia
- Año de publicación
- 2015
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like growth factor (IGF)-1 participation. Therefore, the main interest of this work was to investigate the IGF-1 effect on the extracellular proteolytic activity in MGCs.Methods: Cell culture supernatants and cell lysates of the human MGC line MIO-M1 stimulated with IGF-1 were analyzed for MMP-2 by zymographic and Western blot analysis. The MGCs´ motility was evaluated by scratch wound assay. The MMP-2, β1-integrin, and focal adhesions were detected by confocal microscopy. The localization of active MMPs and actin cytoskeleton were evaluated by in situ zymography.Results: The IGF-1 induced the activation of canonical signaling pathways through the IGF-1R phosphorylation. Culture supernatants showed a relative decrease in the active form of MMP-2, correlating with an increased accumulation of MMP-2 protein in the MGCs´ lysate. The IGF-1 effect on MMP-2 was abolished by an IGF-1R blocking antibody, αIR3, as well as by the PI3-kinase inhibitor, LY294002. The IGF-1 increased the migratory capacity of MGCs, which was blocked by the GM6001 MMP inhibitor, LY294002 and αIR3. Finally, IGF-1 induced the intracellular distribution of MMP-2 toward cellular protrusions and the partial colocalization with β1-integrin and phospo-focal adhesion kinase signals. Gelatinase activity was concentrated along F-actin filaments.Conclusions: Taken together, these data indicate that IGF-1, through its receptor activation, regulates MGCs´ motility by a mechanism that involves the MMP-2 and PI3K signaling pathway.
Fil: Lorenc, Valeria Erika. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Jaldín Fincati, Javier Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Luna Pinto, Jose Domingo. Departamento de Vítreo-retina, Centro Privado de Ojos R; Argentina
Fil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Sanchez, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina - Materia
-
RETINA
GLIA
PROTEASES
MIGRATION - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/46437
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oai:ri.conicet.gov.ar:11336/46437 |
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network_name_str |
CONICET Digital (CONICET) |
spelling |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motilityLorenc, Valeria ErikaJaldín Fincati, Javier RobertoLuna Pinto, Jose DomingoChiabrando, Gustavo AlbertoSanchez, Maria CeciliaRETINAGLIAPROTEASESMIGRATIONhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like growth factor (IGF)-1 participation. Therefore, the main interest of this work was to investigate the IGF-1 effect on the extracellular proteolytic activity in MGCs.Methods: Cell culture supernatants and cell lysates of the human MGC line MIO-M1 stimulated with IGF-1 were analyzed for MMP-2 by zymographic and Western blot analysis. The MGCs´ motility was evaluated by scratch wound assay. The MMP-2, β1-integrin, and focal adhesions were detected by confocal microscopy. The localization of active MMPs and actin cytoskeleton were evaluated by in situ zymography.Results: The IGF-1 induced the activation of canonical signaling pathways through the IGF-1R phosphorylation. Culture supernatants showed a relative decrease in the active form of MMP-2, correlating with an increased accumulation of MMP-2 protein in the MGCs´ lysate. The IGF-1 effect on MMP-2 was abolished by an IGF-1R blocking antibody, αIR3, as well as by the PI3-kinase inhibitor, LY294002. The IGF-1 increased the migratory capacity of MGCs, which was blocked by the GM6001 MMP inhibitor, LY294002 and αIR3. Finally, IGF-1 induced the intracellular distribution of MMP-2 toward cellular protrusions and the partial colocalization with β1-integrin and phospo-focal adhesion kinase signals. Gelatinase activity was concentrated along F-actin filaments.Conclusions: Taken together, these data indicate that IGF-1, through its receptor activation, regulates MGCs´ motility by a mechanism that involves the MMP-2 and PI3K signaling pathway.Fil: Lorenc, Valeria Erika. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Jaldín Fincati, Javier Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Luna Pinto, Jose Domingo. Departamento de Vítreo-retina, Centro Privado de Ojos R; ArgentinaFil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Sanchez, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaAssociation for Research in Vision and Ophthalmology2015-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/46437Lorenc, Valeria Erika; Jaldín Fincati, Javier Roberto; Luna Pinto, Jose Domingo; Chiabrando, Gustavo Alberto; Sanchez, Maria Cecilia; IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility; Association for Research in Vision and Ophthalmology; Investigative Ophthalmology & Visual Science; 56; 11; 10-20150146-04041552-5783CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pubmed/26513500info:eu-repo/semantics/altIdentifier/doi/10.1167/iovs.15-17496info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:04:35Zoai:ri.conicet.gov.ar:11336/46437instacron: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 10:04:35.414CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
title |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
spellingShingle |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility Lorenc, Valeria Erika RETINA GLIA PROTEASES MIGRATION |
title_short |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
title_full |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
title_fullStr |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
title_full_unstemmed |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
title_sort |
IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility |
dc.creator.none.fl_str_mv |
Lorenc, Valeria Erika Jaldín Fincati, Javier Roberto Luna Pinto, Jose Domingo Chiabrando, Gustavo Alberto Sanchez, Maria Cecilia |
author |
Lorenc, Valeria Erika |
author_facet |
Lorenc, Valeria Erika Jaldín Fincati, Javier Roberto Luna Pinto, Jose Domingo Chiabrando, Gustavo Alberto Sanchez, Maria Cecilia |
author_role |
author |
author2 |
Jaldín Fincati, Javier Roberto Luna Pinto, Jose Domingo Chiabrando, Gustavo Alberto Sanchez, Maria Cecilia |
author2_role |
author author author author |
dc.subject.none.fl_str_mv |
RETINA GLIA PROTEASES MIGRATION |
topic |
RETINA GLIA PROTEASES MIGRATION |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/3.1 https://purl.org/becyt/ford/3 |
dc.description.none.fl_txt_mv |
In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like growth factor (IGF)-1 participation. Therefore, the main interest of this work was to investigate the IGF-1 effect on the extracellular proteolytic activity in MGCs.Methods: Cell culture supernatants and cell lysates of the human MGC line MIO-M1 stimulated with IGF-1 were analyzed for MMP-2 by zymographic and Western blot analysis. The MGCs´ motility was evaluated by scratch wound assay. The MMP-2, β1-integrin, and focal adhesions were detected by confocal microscopy. The localization of active MMPs and actin cytoskeleton were evaluated by in situ zymography.Results: The IGF-1 induced the activation of canonical signaling pathways through the IGF-1R phosphorylation. Culture supernatants showed a relative decrease in the active form of MMP-2, correlating with an increased accumulation of MMP-2 protein in the MGCs´ lysate. The IGF-1 effect on MMP-2 was abolished by an IGF-1R blocking antibody, αIR3, as well as by the PI3-kinase inhibitor, LY294002. The IGF-1 increased the migratory capacity of MGCs, which was blocked by the GM6001 MMP inhibitor, LY294002 and αIR3. Finally, IGF-1 induced the intracellular distribution of MMP-2 toward cellular protrusions and the partial colocalization with β1-integrin and phospo-focal adhesion kinase signals. Gelatinase activity was concentrated along F-actin filaments.Conclusions: Taken together, these data indicate that IGF-1, through its receptor activation, regulates MGCs´ motility by a mechanism that involves the MMP-2 and PI3K signaling pathway. Fil: Lorenc, Valeria Erika. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Jaldín Fincati, Javier Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Luna Pinto, Jose Domingo. Departamento de Vítreo-retina, Centro Privado de Ojos R; Argentina Fil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Sanchez, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina |
description |
In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like growth factor (IGF)-1 participation. Therefore, the main interest of this work was to investigate the IGF-1 effect on the extracellular proteolytic activity in MGCs.Methods: Cell culture supernatants and cell lysates of the human MGC line MIO-M1 stimulated with IGF-1 were analyzed for MMP-2 by zymographic and Western blot analysis. The MGCs´ motility was evaluated by scratch wound assay. The MMP-2, β1-integrin, and focal adhesions were detected by confocal microscopy. The localization of active MMPs and actin cytoskeleton were evaluated by in situ zymography.Results: The IGF-1 induced the activation of canonical signaling pathways through the IGF-1R phosphorylation. Culture supernatants showed a relative decrease in the active form of MMP-2, correlating with an increased accumulation of MMP-2 protein in the MGCs´ lysate. The IGF-1 effect on MMP-2 was abolished by an IGF-1R blocking antibody, αIR3, as well as by the PI3-kinase inhibitor, LY294002. The IGF-1 increased the migratory capacity of MGCs, which was blocked by the GM6001 MMP inhibitor, LY294002 and αIR3. Finally, IGF-1 induced the intracellular distribution of MMP-2 toward cellular protrusions and the partial colocalization with β1-integrin and phospo-focal adhesion kinase signals. Gelatinase activity was concentrated along F-actin filaments.Conclusions: Taken together, these data indicate that IGF-1, through its receptor activation, regulates MGCs´ motility by a mechanism that involves the MMP-2 and PI3K signaling pathway. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-10 |
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/11336/46437 Lorenc, Valeria Erika; Jaldín Fincati, Javier Roberto; Luna Pinto, Jose Domingo; Chiabrando, Gustavo Alberto; Sanchez, Maria Cecilia; IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility; Association for Research in Vision and Ophthalmology; Investigative Ophthalmology & Visual Science; 56; 11; 10-2015 0146-0404 1552-5783 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/46437 |
identifier_str_mv |
Lorenc, Valeria Erika; Jaldín Fincati, Javier Roberto; Luna Pinto, Jose Domingo; Chiabrando, Gustavo Alberto; Sanchez, Maria Cecilia; IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility; Association for Research in Vision and Ophthalmology; Investigative Ophthalmology & Visual Science; 56; 11; 10-2015 0146-0404 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://www.ncbi.nlm.nih.gov/pubmed/26513500 info:eu-repo/semantics/altIdentifier/doi/10.1167/iovs.15-17496 |
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 |
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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1844613873596891136 |
score |
13.070432 |