Cargando…

High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy

To investigate whether high glucose (HG) alters Rab20 expression and compromises gap junction intercellular communication (GJIC) and cell survival, retinal cells were studied for altered intracellular trafficking of connexin 43 (Cx43). Retinal endothelial cells (RRECs) and retinal Müller cells (rMCs...

Descripción completa

Detalles Bibliográficos
Autores principales: Kim, Dongjoon, Lewis, Casey Stottrup, Sarthy, Vijay P., Roy, Sayon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7699280/
https://www.ncbi.nlm.nih.gov/pubmed/33227912
http://dx.doi.org/10.3390/jcm9113710
_version_ 1783616013477085184
author Kim, Dongjoon
Lewis, Casey Stottrup
Sarthy, Vijay P.
Roy, Sayon
author_facet Kim, Dongjoon
Lewis, Casey Stottrup
Sarthy, Vijay P.
Roy, Sayon
author_sort Kim, Dongjoon
collection PubMed
description To investigate whether high glucose (HG) alters Rab20 expression and compromises gap junction intercellular communication (GJIC) and cell survival, retinal cells were studied for altered intracellular trafficking of connexin 43 (Cx43). Retinal endothelial cells (RRECs) and retinal Müller cells (rMCs) were grown in normal (N; 5 mM glucose) or HG (30 mM glucose) medium for seven days. In parallel, cells grown in HG medium were transfected with either Rab20 siRNA or scrambled siRNA as a control. Rab20 and Cx43 expression and their localization and distribution were assessed using Western Blot and immunostaining, respectively. Changes in GJIC activity were assessed using scrape load dye transfer, and apoptosis was identified using differential dye staining assay. In RRECs or rMCs grown in HG medium, Rab20 expression was significantly increased concomitant with a decreased number of Cx43 plaques. Importantly, a significant increase in the number of Cx43 plaques and GJIC activity was observed in cells transfected with Rab20 siRNA. Additionally, Rab20 downregulation inhibited HG-induced apoptosis in RRECs and rMCs. Results indicate HG-mediated Rab20 upregulation decreases Cx43 localization at the cell surface, resulting in compromised GJIC activity. Reducing Rab20 expression could be a useful strategy in preventing HG-induced vascular and Müller cell death associated with diabetic retinopathy.
format Online
Article
Text
id pubmed-7699280
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-76992802020-11-29 High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy Kim, Dongjoon Lewis, Casey Stottrup Sarthy, Vijay P. Roy, Sayon J Clin Med Article To investigate whether high glucose (HG) alters Rab20 expression and compromises gap junction intercellular communication (GJIC) and cell survival, retinal cells were studied for altered intracellular trafficking of connexin 43 (Cx43). Retinal endothelial cells (RRECs) and retinal Müller cells (rMCs) were grown in normal (N; 5 mM glucose) or HG (30 mM glucose) medium for seven days. In parallel, cells grown in HG medium were transfected with either Rab20 siRNA or scrambled siRNA as a control. Rab20 and Cx43 expression and their localization and distribution were assessed using Western Blot and immunostaining, respectively. Changes in GJIC activity were assessed using scrape load dye transfer, and apoptosis was identified using differential dye staining assay. In RRECs or rMCs grown in HG medium, Rab20 expression was significantly increased concomitant with a decreased number of Cx43 plaques. Importantly, a significant increase in the number of Cx43 plaques and GJIC activity was observed in cells transfected with Rab20 siRNA. Additionally, Rab20 downregulation inhibited HG-induced apoptosis in RRECs and rMCs. Results indicate HG-mediated Rab20 upregulation decreases Cx43 localization at the cell surface, resulting in compromised GJIC activity. Reducing Rab20 expression could be a useful strategy in preventing HG-induced vascular and Müller cell death associated with diabetic retinopathy. MDPI 2020-11-19 /pmc/articles/PMC7699280/ /pubmed/33227912 http://dx.doi.org/10.3390/jcm9113710 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kim, Dongjoon
Lewis, Casey Stottrup
Sarthy, Vijay P.
Roy, Sayon
High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title_full High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title_fullStr High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title_full_unstemmed High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title_short High-Glucose-Induced Rab20 Upregulation Disrupts Gap Junction Intercellular Communication and Promotes Apoptosis in Retinal Endothelial and Müller Cells: Implications for Diabetic Retinopathy
title_sort high-glucose-induced rab20 upregulation disrupts gap junction intercellular communication and promotes apoptosis in retinal endothelial and müller cells: implications for diabetic retinopathy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7699280/
https://www.ncbi.nlm.nih.gov/pubmed/33227912
http://dx.doi.org/10.3390/jcm9113710
work_keys_str_mv AT kimdongjoon highglucoseinducedrab20upregulationdisruptsgapjunctionintercellularcommunicationandpromotesapoptosisinretinalendothelialandmullercellsimplicationsfordiabeticretinopathy
AT lewiscaseystottrup highglucoseinducedrab20upregulationdisruptsgapjunctionintercellularcommunicationandpromotesapoptosisinretinalendothelialandmullercellsimplicationsfordiabeticretinopathy
AT sarthyvijayp highglucoseinducedrab20upregulationdisruptsgapjunctionintercellularcommunicationandpromotesapoptosisinretinalendothelialandmullercellsimplicationsfordiabeticretinopathy
AT roysayon highglucoseinducedrab20upregulationdisruptsgapjunctionintercellularcommunicationandpromotesapoptosisinretinalendothelialandmullercellsimplicationsfordiabeticretinopathy