Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target

Autores
Mader, Julieta Ailen; Fernandez Delias, María Florencia; Chrestia, Juan Facundo; Esandi, María del Carmen; Bouzat, Cecilia Beatriz
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The α7 nicotinic acetylcholine receptor is highly expressed in the brain and is also present in non-neuronal cells, including immune and epithelial cells. It is involved in cognition, memory, pain, neuroprotection and inflammation and its potentiation has emerged as a therapeutic strategy for neurological, neurodegenerative, and inflammatory disorders. Given that the increase in oxidative stress in retinal pigment epithelial cells contributes to the development of age-related macular degeneration and that α7 activation exerts cell protective effects, we explored the presence and functional relevance of α7 in D407 retinal pigment epithelium cells, a model system for various retinal diseases. By real time PCR, and indirect immunofluorescence using confocalmicroscopy and flow cytometry we demonstrated the presence of α7 in these epithelial cells. To determine the presence of functional receptors, we measured the movement of intracellular calcium levels triggered by the activation of α7. A pulse of ACh together with an α7 positive allosteric modulator revealed a 3-fold increase in intracellular calcium measured with the fluo-3AM probe. To mimic the events occurring in age-related macular degeneration, we treated cells with ferric ammonium citrate (FAC) to induce stress damage andmeasured reactive oxygen species (ROS) with the fluorescent probe DCFH-DA. FAC treatment resulted in a significant increase in ROS levels with respect to the control. To determine if α7 protects against oxidative damage, we exposed cells to a specific α7 agonist, PNU-282987, before the FAC treatment. Notably, PNU-282987 exhibited protective effects against the damage, leading to a reduction in ROS levels compared to the treated cells. Overall, by identifying for the first time the presence of α7 in the D407 cell line and revealing its protective role against oxidative damage, we propose α7 as a promising therapeutic target for retinal neurodegenerative disorders.
Fil: Mader, Julieta Ailen. 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: Fernandez Delias, María Florencia. 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: Chrestia, Juan Facundo. 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: Esandi, María del Carmen. 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: Bouzat, Cecilia Beatriz. 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
LVI Reunión Anual De La Asociación Argentina De Farmacología Experimental
Bahía Blanca
Argentina
Asociación Argentina De Farmacología Experimental
Materia
LIGAND-GATED ION CHANNEL
NICOTINIC RECEPTOR
EPITHELIAL CELLS
ALPHA7
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:11336/278078

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network_name_str CONICET Digital (CONICET)
spelling Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic TargetMader, Julieta AilenFernandez Delias, María FlorenciaChrestia, Juan FacundoEsandi, María del CarmenBouzat, Cecilia BeatrizLIGAND-GATED ION CHANNELNICOTINIC RECEPTOREPITHELIAL CELLSALPHA7https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The α7 nicotinic acetylcholine receptor is highly expressed in the brain and is also present in non-neuronal cells, including immune and epithelial cells. It is involved in cognition, memory, pain, neuroprotection and inflammation and its potentiation has emerged as a therapeutic strategy for neurological, neurodegenerative, and inflammatory disorders. Given that the increase in oxidative stress in retinal pigment epithelial cells contributes to the development of age-related macular degeneration and that α7 activation exerts cell protective effects, we explored the presence and functional relevance of α7 in D407 retinal pigment epithelium cells, a model system for various retinal diseases. By real time PCR, and indirect immunofluorescence using confocalmicroscopy and flow cytometry we demonstrated the presence of α7 in these epithelial cells. To determine the presence of functional receptors, we measured the movement of intracellular calcium levels triggered by the activation of α7. A pulse of ACh together with an α7 positive allosteric modulator revealed a 3-fold increase in intracellular calcium measured with the fluo-3AM probe. To mimic the events occurring in age-related macular degeneration, we treated cells with ferric ammonium citrate (FAC) to induce stress damage andmeasured reactive oxygen species (ROS) with the fluorescent probe DCFH-DA. FAC treatment resulted in a significant increase in ROS levels with respect to the control. To determine if α7 protects against oxidative damage, we exposed cells to a specific α7 agonist, PNU-282987, before the FAC treatment. Notably, PNU-282987 exhibited protective effects against the damage, leading to a reduction in ROS levels compared to the treated cells. Overall, by identifying for the first time the presence of α7 in the D407 cell line and revealing its protective role against oxidative damage, we propose α7 as a promising therapeutic target for retinal neurodegenerative disorders.Fil: Mader, Julieta Ailen. 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: Fernandez Delias, María Florencia. 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: Chrestia, Juan Facundo. 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: Esandi, María del Carmen. 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: Bouzat, Cecilia Beatriz. 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; ArgentinaLVI Reunión Anual De La Asociación Argentina De Farmacología ExperimentalBahía BlancaArgentinaAsociación Argentina De Farmacología ExperimentalAsociación Argentina De Farmacología Experimental2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/278078Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target; LVI Reunión Anual De La Asociación Argentina De Farmacología Experimental; Bahía Blanca; Argentina; 2024; 108-108978-631-90806-0-5CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://aafeargentina.org/congresos-aafe/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-23T13:27:43Zoai:ri.conicet.gov.ar:11336/278078instacron: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:34982025-12-23 13:27:44.113CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
title Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
spellingShingle Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
Mader, Julieta Ailen
LIGAND-GATED ION CHANNEL
NICOTINIC RECEPTOR
EPITHELIAL CELLS
ALPHA7
title_short Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
title_full Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
title_fullStr Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
title_full_unstemmed Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
title_sort Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target
dc.creator.none.fl_str_mv Mader, Julieta Ailen
Fernandez Delias, María Florencia
Chrestia, Juan Facundo
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author Mader, Julieta Ailen
author_facet Mader, Julieta Ailen
Fernandez Delias, María Florencia
Chrestia, Juan Facundo
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author_role author
author2 Fernandez Delias, María Florencia
Chrestia, Juan Facundo
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author2_role author
author
author
author
dc.subject.none.fl_str_mv LIGAND-GATED ION CHANNEL
NICOTINIC RECEPTOR
EPITHELIAL CELLS
ALPHA7
topic LIGAND-GATED ION CHANNEL
NICOTINIC RECEPTOR
EPITHELIAL CELLS
ALPHA7
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The α7 nicotinic acetylcholine receptor is highly expressed in the brain and is also present in non-neuronal cells, including immune and epithelial cells. It is involved in cognition, memory, pain, neuroprotection and inflammation and its potentiation has emerged as a therapeutic strategy for neurological, neurodegenerative, and inflammatory disorders. Given that the increase in oxidative stress in retinal pigment epithelial cells contributes to the development of age-related macular degeneration and that α7 activation exerts cell protective effects, we explored the presence and functional relevance of α7 in D407 retinal pigment epithelium cells, a model system for various retinal diseases. By real time PCR, and indirect immunofluorescence using confocalmicroscopy and flow cytometry we demonstrated the presence of α7 in these epithelial cells. To determine the presence of functional receptors, we measured the movement of intracellular calcium levels triggered by the activation of α7. A pulse of ACh together with an α7 positive allosteric modulator revealed a 3-fold increase in intracellular calcium measured with the fluo-3AM probe. To mimic the events occurring in age-related macular degeneration, we treated cells with ferric ammonium citrate (FAC) to induce stress damage andmeasured reactive oxygen species (ROS) with the fluorescent probe DCFH-DA. FAC treatment resulted in a significant increase in ROS levels with respect to the control. To determine if α7 protects against oxidative damage, we exposed cells to a specific α7 agonist, PNU-282987, before the FAC treatment. Notably, PNU-282987 exhibited protective effects against the damage, leading to a reduction in ROS levels compared to the treated cells. Overall, by identifying for the first time the presence of α7 in the D407 cell line and revealing its protective role against oxidative damage, we propose α7 as a promising therapeutic target for retinal neurodegenerative disorders.
Fil: Mader, Julieta Ailen. 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: Fernandez Delias, María Florencia. 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: Chrestia, Juan Facundo. 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: Esandi, María del Carmen. 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: Bouzat, Cecilia Beatriz. 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
LVI Reunión Anual De La Asociación Argentina De Farmacología Experimental
Bahía Blanca
Argentina
Asociación Argentina De Farmacología Experimental
description The α7 nicotinic acetylcholine receptor is highly expressed in the brain and is also present in non-neuronal cells, including immune and epithelial cells. It is involved in cognition, memory, pain, neuroprotection and inflammation and its potentiation has emerged as a therapeutic strategy for neurological, neurodegenerative, and inflammatory disorders. Given that the increase in oxidative stress in retinal pigment epithelial cells contributes to the development of age-related macular degeneration and that α7 activation exerts cell protective effects, we explored the presence and functional relevance of α7 in D407 retinal pigment epithelium cells, a model system for various retinal diseases. By real time PCR, and indirect immunofluorescence using confocalmicroscopy and flow cytometry we demonstrated the presence of α7 in these epithelial cells. To determine the presence of functional receptors, we measured the movement of intracellular calcium levels triggered by the activation of α7. A pulse of ACh together with an α7 positive allosteric modulator revealed a 3-fold increase in intracellular calcium measured with the fluo-3AM probe. To mimic the events occurring in age-related macular degeneration, we treated cells with ferric ammonium citrate (FAC) to induce stress damage andmeasured reactive oxygen species (ROS) with the fluorescent probe DCFH-DA. FAC treatment resulted in a significant increase in ROS levels with respect to the control. To determine if α7 protects against oxidative damage, we exposed cells to a specific α7 agonist, PNU-282987, before the FAC treatment. Notably, PNU-282987 exhibited protective effects against the damage, leading to a reduction in ROS levels compared to the treated cells. Overall, by identifying for the first time the presence of α7 in the D407 cell line and revealing its protective role against oxidative damage, we propose α7 as a promising therapeutic target for retinal neurodegenerative disorders.
publishDate 2024
dc.date.none.fl_str_mv 2024
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info:eu-repo/semantics/conferenceObject
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http://purl.org/coar/resource_type/c_5794
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dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/278078
Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target; LVI Reunión Anual De La Asociación Argentina De Farmacología Experimental; Bahía Blanca; Argentina; 2024; 108-108
978-631-90806-0-5
CONICET Digital
CONICET
url http://hdl.handle.net/11336/278078
identifier_str_mv Exploring The α7 Nicotinic Receptor In Human Retinal Pigment Epithelium Cells As A Novel Therapeutic Target; LVI Reunión Anual De La Asociación Argentina De Farmacología Experimental; Bahía Blanca; Argentina; 2024; 108-108
978-631-90806-0-5
CONICET Digital
CONICET
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language eng
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publisher.none.fl_str_mv Asociación Argentina De Farmacología Experimental
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