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Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor

PURPOSE: The purpose of this study was to investigate responses to toxic cellular stresses in different human ocular epithelia. METHODS: Reactivity with a specific anti-P2X7 antibody was studied using confocal fluorescence microscopy on conjunctival, corneal, lens, and retinal cell lines as well as...

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Autores principales: Dutot, Mélody, Liang, Hong, Pauloin, Thierry, Brignole-Baudouin, Françoise, Baudouin, Christophe, Warnet, Jean-Michel, Rat, Patrice
Formato: Texto
Lenguaje:English
Publicado: Molecular Vision 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2386509/
https://www.ncbi.nlm.nih.gov/pubmed/18490962
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author Dutot, Mélody
Liang, Hong
Pauloin, Thierry
Brignole-Baudouin, Françoise
Baudouin, Christophe
Warnet, Jean-Michel
Rat, Patrice
author_facet Dutot, Mélody
Liang, Hong
Pauloin, Thierry
Brignole-Baudouin, Françoise
Baudouin, Christophe
Warnet, Jean-Michel
Rat, Patrice
author_sort Dutot, Mélody
collection PubMed
description PURPOSE: The purpose of this study was to investigate responses to toxic cellular stresses in different human ocular epithelia. METHODS: Reactivity with a specific anti-P2X7 antibody was studied using confocal fluorescence microscopy on conjunctival, corneal, lens, and retinal cell lines as well as using impression cytology on human ocular cells. Activation of the P2X7 receptor by selective agonists (ATP and benzoylbenzoyl-ATP) and inhibition by antagonists (oATP, KN-62, and PPADS) were evaluated using the quinolinium,4-[(3-methyl-2-(3H)-benzoxazolylidene) methyl]-1-[3-(triethylammonio)propyl]di-iodide (YO-PRO-1) test in cytofluorometry. Different specific stresses were then induced by a chemical toxin (benzalkonium chloride) and a chemical oxidant (tert-butyl hydroperoxide) to assess the role of the P2X7 receptor. Modulation of P2X7 receptor activation was performed with several ionic solutions. RESULTS: Our data show that four cell lines express the P2X7 cell death purinergic receptor as judged by reactivity with a specific anti-P2X7 antibody, activation by the selective P2X7 agonist benzoylbenzoyl-ATP and to a lesser extent by ATP (YO-PRO-1 dye uptake), and inhibition by three antagonists (oATP, KN-62, and PPADS). Benzalkonium chloride, a widely used preservative, induced dramatic membrane permeabilization through P2X7 pore opening on conjunctival and corneal epithelia. Reactive oxygen species, induced by tert-butyl hydroperoxide, lead to P2X7 receptor activation on retinal pigment epithelium. Modulation of P2X7 receptor activation was obtained with extracellular Ca(2+) and Mg(2+) and with a controlled ionization marine solution rich in different divalent cations. This marine solution could be proposed as a new ophthalmic solution. CONCLUSIONS: Our observations reveal a novel pathway for epithelial cells apoptosis/cytolysis by inducing different toxic stresses and their modulation by using ionic solutions.
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spelling pubmed-23865092008-05-19 Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor Dutot, Mélody Liang, Hong Pauloin, Thierry Brignole-Baudouin, Françoise Baudouin, Christophe Warnet, Jean-Michel Rat, Patrice Mol Vis Research Article PURPOSE: The purpose of this study was to investigate responses to toxic cellular stresses in different human ocular epithelia. METHODS: Reactivity with a specific anti-P2X7 antibody was studied using confocal fluorescence microscopy on conjunctival, corneal, lens, and retinal cell lines as well as using impression cytology on human ocular cells. Activation of the P2X7 receptor by selective agonists (ATP and benzoylbenzoyl-ATP) and inhibition by antagonists (oATP, KN-62, and PPADS) were evaluated using the quinolinium,4-[(3-methyl-2-(3H)-benzoxazolylidene) methyl]-1-[3-(triethylammonio)propyl]di-iodide (YO-PRO-1) test in cytofluorometry. Different specific stresses were then induced by a chemical toxin (benzalkonium chloride) and a chemical oxidant (tert-butyl hydroperoxide) to assess the role of the P2X7 receptor. Modulation of P2X7 receptor activation was performed with several ionic solutions. RESULTS: Our data show that four cell lines express the P2X7 cell death purinergic receptor as judged by reactivity with a specific anti-P2X7 antibody, activation by the selective P2X7 agonist benzoylbenzoyl-ATP and to a lesser extent by ATP (YO-PRO-1 dye uptake), and inhibition by three antagonists (oATP, KN-62, and PPADS). Benzalkonium chloride, a widely used preservative, induced dramatic membrane permeabilization through P2X7 pore opening on conjunctival and corneal epithelia. Reactive oxygen species, induced by tert-butyl hydroperoxide, lead to P2X7 receptor activation on retinal pigment epithelium. Modulation of P2X7 receptor activation was obtained with extracellular Ca(2+) and Mg(2+) and with a controlled ionization marine solution rich in different divalent cations. This marine solution could be proposed as a new ophthalmic solution. CONCLUSIONS: Our observations reveal a novel pathway for epithelial cells apoptosis/cytolysis by inducing different toxic stresses and their modulation by using ionic solutions. Molecular Vision 2008-05-19 /pmc/articles/PMC2386509/ /pubmed/18490962 Text en http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Dutot, Mélody
Liang, Hong
Pauloin, Thierry
Brignole-Baudouin, Françoise
Baudouin, Christophe
Warnet, Jean-Michel
Rat, Patrice
Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title_full Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title_fullStr Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title_full_unstemmed Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title_short Effects of toxic cellular stresses and divalent cations on the human P2X7 cell death receptor
title_sort effects of toxic cellular stresses and divalent cations on the human p2x7 cell death receptor
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2386509/
https://www.ncbi.nlm.nih.gov/pubmed/18490962
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