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Title (Primary) Oxygen/glucose deprivation increases the integration of recombinant P2X(7) receptors into the plasma membrane of HEK293 cells
Author Milius, D.; Gröger-Arndt, H.; Stanchev, D.; Lange-Dohna, C.; Rossner, S.; Sperlágh, B.; Wirkner, K.; Illes, P.;
Journal Toxicology
Year 2007
Department PROTEOM;
Volume 238
Issue 1
Language englisch;
Abstract Recombinant human P2X(7) receptors, C-terminally labelled with enhanced green fluorescent protein (P2X(7)-EGFP), were transiently expressed in HEK293 cells. Activation of these receptors by their preferential agonist 2',3'-O-(4-benzoylbenzoyl)-ATP (BzATP) induced inward currents and propidium ion uptake indicating the opening of cationic channels and of large pores penneable for dye molecules, respectively. Two mutants of P2X(7) receptors (P2X(7)-EGFP-I568N, -E496A) representing polymorphisms in the P2X(7) gene known to interfere with normal receptor-trafficking and with optimal assembly of its subunits, responded with much lower current amplitudes to BzATP than their wild-type counterpart. Similarly, the normal propidium ion uptake induced by BzATP at the wild-type P2X(7) receptor was abolished by the two mutants. Confocal laser scanning microscopy indicated that in vitro ischernia of 12 h duration increased the integration of P2X(7)-EGFP, but not of its two mutants, into the plasma membrane of HEK293 cells. Further, this ischernic stimulus facilitated the current response to BzATP in HEK293 cells permanently transfected with P2X(7) receptors. Finally, the fluorescence intensity per cell measured by flow cytometry and P2X(7) antibodies directed against an extracellular, but not an intracellular epitope of the receptor, were also increased. In conclusion, P2X(7) receptors may alter their trafficking properties during ischernia and thereby contribute to the ATP-induced damage of various cell-types including neurons. (c) 2007 Elsevier Ireland Ltd. All rights reserved
ID 2077
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=2077
Milius, D., Gröger-Arndt, H., Stanchev, D., Lange-Dohna, C., Rossner, S., Sperlágh, B., Wirkner, K., Illes, P. (2007):
Oxygen/glucose deprivation increases the integration of recombinant P2X(7) receptors into the plasma membrane of HEK293 cells
Toxicology 238 (1), 60 - 69