A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells

Autores
Morera, Luis Pedro; Díaz, Nicolás Maximiliano; Guido, Mario Eduardo
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The retina is a heterogeneous tissue composed of different neuronal cells (photoreceptor (PRC), horizontal (HC), amacrine, bipolar and retinal ganglion RGC cells) and glial cells. Our aim in this work was to purify HCs from the chicken embryonic retina to obtain primary cultures highly enriched in these cells for further characterization. To this end, disaggregated retinas of chicken embryos at day 15 were subjected to a discontinuous bovine serum albumin (BSA) gradient of concentrations raging from 1 to 4%. After centrifugation, cells collected from the different phases were cultured for 4 days and characterized by immunochemistry and cell morphology. Phases were examined with specific antibodies against HC markers such as PROX-1, Islet-1 and calretinin, together with markers for other retinal cell populations. The results show that only in the fraction corresponding to 2.5% BSA did most of the cells display PROX-1 and Islet-1 positive immunoreactivities with a typical HC morphology. Moreover, Western blot assays indicate that the 2.5 % BSA phase exhibits the strongest PROX-1 immunolabeling, denoting the typical molecular weight (MW) of ~ 83KDa. Based on an accurate morphological analysis, a number of cells in this fraction resembled H1- and H3-type HCs (axon-bearing "brush-shaped" and axon-less "candelabrum-shaped" HCs respectively). In conclusion, the BSA gradient has so far proved to be a simple yet very useful method to selectively separate specific retinal cell types for the further study and characterization of their molecular, biochemical and electrophysiological properties.
Fil: Morera, Luis Pedro. Universidad Empresarial Siglo XXI. Vicerrectorado de Innovación e Investigación. Instituto de Organizaciones Saludables; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Díaz, Nicolás Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Guido, Mario Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Materia
DEVELOPMENT
HORIZONTAL CELLS
ISLET
PROX1
RETINA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/197567
Acceder
id CONICETDig_66bc2d24274f405b0dbd50ee5da3b929
oai_identifier_str oai:ri.conicet.gov.ar:11336/197567
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cellsMorera, Luis PedroDíaz, Nicolás MaximilianoGuido, Mario EduardoDEVELOPMENTHORIZONTAL CELLSISLETPROX1RETINAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The retina is a heterogeneous tissue composed of different neuronal cells (photoreceptor (PRC), horizontal (HC), amacrine, bipolar and retinal ganglion RGC cells) and glial cells. Our aim in this work was to purify HCs from the chicken embryonic retina to obtain primary cultures highly enriched in these cells for further characterization. To this end, disaggregated retinas of chicken embryos at day 15 were subjected to a discontinuous bovine serum albumin (BSA) gradient of concentrations raging from 1 to 4%. After centrifugation, cells collected from the different phases were cultured for 4 days and characterized by immunochemistry and cell morphology. Phases were examined with specific antibodies against HC markers such as PROX-1, Islet-1 and calretinin, together with markers for other retinal cell populations. The results show that only in the fraction corresponding to 2.5% BSA did most of the cells display PROX-1 and Islet-1 positive immunoreactivities with a typical HC morphology. Moreover, Western blot assays indicate that the 2.5 % BSA phase exhibits the strongest PROX-1 immunolabeling, denoting the typical molecular weight (MW) of ~ 83KDa. Based on an accurate morphological analysis, a number of cells in this fraction resembled H1- and H3-type HCs (axon-bearing "brush-shaped" and axon-less "candelabrum-shaped" HCs respectively). In conclusion, the BSA gradient has so far proved to be a simple yet very useful method to selectively separate specific retinal cell types for the further study and characterization of their molecular, biochemical and electrophysiological properties.Fil: Morera, Luis Pedro. Universidad Empresarial Siglo XXI. Vicerrectorado de Innovación e Investigación. Instituto de Organizaciones Saludables; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Díaz, Nicolás Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Guido, Mario Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaAcademic Press Ltd - Elsevier Science Ltd2012-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/197567Morera, Luis Pedro; Díaz, Nicolás Maximiliano; Guido, Mario Eduardo; A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells; Academic Press Ltd - Elsevier Science Ltd; Experimental Eye Research; 101; 8-2012; 44-480014-4835CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.exer.2012.05.010info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S001448351200156Xinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:10:23Zoai:ri.conicet.gov.ar:11336/197567instacron: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:34982025-09-10 13:10:24.137CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
title A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
spellingShingle A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
Morera, Luis Pedro
DEVELOPMENT
HORIZONTAL CELLS
ISLET
PROX1
RETINA
title_short A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
title_full A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
title_fullStr A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
title_full_unstemmed A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
title_sort A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells
dc.creator.none.fl_str_mv Morera, Luis Pedro
Díaz, Nicolás Maximiliano
Guido, Mario Eduardo
author Morera, Luis Pedro
author_facet Morera, Luis Pedro
Díaz, Nicolás Maximiliano
Guido, Mario Eduardo
author_role author
author2 Díaz, Nicolás Maximiliano
Guido, Mario Eduardo
author2_role author
author
dc.subject.none.fl_str_mv DEVELOPMENT
HORIZONTAL CELLS
ISLET
PROX1
RETINA
topic DEVELOPMENT
HORIZONTAL CELLS
ISLET
PROX1
RETINA
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 retina is a heterogeneous tissue composed of different neuronal cells (photoreceptor (PRC), horizontal (HC), amacrine, bipolar and retinal ganglion RGC cells) and glial cells. Our aim in this work was to purify HCs from the chicken embryonic retina to obtain primary cultures highly enriched in these cells for further characterization. To this end, disaggregated retinas of chicken embryos at day 15 were subjected to a discontinuous bovine serum albumin (BSA) gradient of concentrations raging from 1 to 4%. After centrifugation, cells collected from the different phases were cultured for 4 days and characterized by immunochemistry and cell morphology. Phases were examined with specific antibodies against HC markers such as PROX-1, Islet-1 and calretinin, together with markers for other retinal cell populations. The results show that only in the fraction corresponding to 2.5% BSA did most of the cells display PROX-1 and Islet-1 positive immunoreactivities with a typical HC morphology. Moreover, Western blot assays indicate that the 2.5 % BSA phase exhibits the strongest PROX-1 immunolabeling, denoting the typical molecular weight (MW) of ~ 83KDa. Based on an accurate morphological analysis, a number of cells in this fraction resembled H1- and H3-type HCs (axon-bearing "brush-shaped" and axon-less "candelabrum-shaped" HCs respectively). In conclusion, the BSA gradient has so far proved to be a simple yet very useful method to selectively separate specific retinal cell types for the further study and characterization of their molecular, biochemical and electrophysiological properties.
Fil: Morera, Luis Pedro. Universidad Empresarial Siglo XXI. Vicerrectorado de Innovación e Investigación. Instituto de Organizaciones Saludables; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Díaz, Nicolás Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Guido, Mario Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
description The retina is a heterogeneous tissue composed of different neuronal cells (photoreceptor (PRC), horizontal (HC), amacrine, bipolar and retinal ganglion RGC cells) and glial cells. Our aim in this work was to purify HCs from the chicken embryonic retina to obtain primary cultures highly enriched in these cells for further characterization. To this end, disaggregated retinas of chicken embryos at day 15 were subjected to a discontinuous bovine serum albumin (BSA) gradient of concentrations raging from 1 to 4%. After centrifugation, cells collected from the different phases were cultured for 4 days and characterized by immunochemistry and cell morphology. Phases were examined with specific antibodies against HC markers such as PROX-1, Islet-1 and calretinin, together with markers for other retinal cell populations. The results show that only in the fraction corresponding to 2.5% BSA did most of the cells display PROX-1 and Islet-1 positive immunoreactivities with a typical HC morphology. Moreover, Western blot assays indicate that the 2.5 % BSA phase exhibits the strongest PROX-1 immunolabeling, denoting the typical molecular weight (MW) of ~ 83KDa. Based on an accurate morphological analysis, a number of cells in this fraction resembled H1- and H3-type HCs (axon-bearing "brush-shaped" and axon-less "candelabrum-shaped" HCs respectively). In conclusion, the BSA gradient has so far proved to be a simple yet very useful method to selectively separate specific retinal cell types for the further study and characterization of their molecular, biochemical and electrophysiological properties.
publishDate 2012
dc.date.none.fl_str_mv 2012-08
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/11336/197567
Morera, Luis Pedro; Díaz, Nicolás Maximiliano; Guido, Mario Eduardo; A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells; Academic Press Ltd - Elsevier Science Ltd; Experimental Eye Research; 101; 8-2012; 44-48
0014-4835
CONICET Digital
CONICET
url http://hdl.handle.net/11336/197567
identifier_str_mv Morera, Luis Pedro; Díaz, Nicolás Maximiliano; Guido, Mario Eduardo; A novel method to prepare highly enriched primary cultures of chicken retinal horizontal cells; Academic Press Ltd - Elsevier Science Ltd; Experimental Eye Research; 101; 8-2012; 44-48
0014-4835
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.exer.2012.05.010
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S001448351200156X
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
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 12.993085

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