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Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye
The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non‐pigmented ciliary epithelium (NPCE)–deri...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5824413/ https://www.ncbi.nlm.nih.gov/pubmed/29411534 http://dx.doi.org/10.1111/jcmm.13505 |
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author | Tabak, Saray Schreiber‐Avissar, Sofia Beit‐Yannai, Elie |
author_facet | Tabak, Saray Schreiber‐Avissar, Sofia Beit‐Yannai, Elie |
author_sort | Tabak, Saray |
collection | PubMed |
description | The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non‐pigmented ciliary epithelium (NPCE)–derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose–response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE‐derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β‐Catenin, Axin2 and LEF1) and protein (β‐Catenin, GSK‐3β) expression using real‐time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β‐Catenin, GSK‐3β, as opposed to exposure to high exosomal concentrations. Pro‐MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE‐ or RPE‐derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration‐dependent at their target site. Specifically in the present study, we described a general dose–response at the gene and MMPs activity and a different dose–response regarding key canonical Wnt proteins expression. |
format | Online Article Text |
id | pubmed-5824413 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58244132018-03-01 Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye Tabak, Saray Schreiber‐Avissar, Sofia Beit‐Yannai, Elie J Cell Mol Med Original Articles The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non‐pigmented ciliary epithelium (NPCE)–derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose–response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE‐derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β‐Catenin, Axin2 and LEF1) and protein (β‐Catenin, GSK‐3β) expression using real‐time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β‐Catenin, GSK‐3β, as opposed to exposure to high exosomal concentrations. Pro‐MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE‐ or RPE‐derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration‐dependent at their target site. Specifically in the present study, we described a general dose–response at the gene and MMPs activity and a different dose–response regarding key canonical Wnt proteins expression. John Wiley and Sons Inc. 2018-02-07 2018-03 /pmc/articles/PMC5824413/ /pubmed/29411534 http://dx.doi.org/10.1111/jcmm.13505 Text en © 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Tabak, Saray Schreiber‐Avissar, Sofia Beit‐Yannai, Elie Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title | Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title_full | Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title_fullStr | Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title_full_unstemmed | Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title_short | Extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
title_sort | extracellular vesicles have variable dose‐dependent effects on cultured draining cells in the eye |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5824413/ https://www.ncbi.nlm.nih.gov/pubmed/29411534 http://dx.doi.org/10.1111/jcmm.13505 |
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