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Establishment of an Assay for P2X7 Receptor-Mediated Cell Death  

Lee, Song-Yi (Department of Life Science, Gwangju Institute of Science and Technology)
Jo, Sooyeon (Department of Life Science, Gwangju Institute of Science and Technology)
Lee, Ga Eun (Department of Life Science, Gwangju Institute of Science and Technology)
Jeong, Lak Shin (College of Pharmacy, Ewha Womans University)
Kim, Yong-Chul (Department of Life Science, Gwangju Institute of Science and Technology)
Park, Chul-Seung (Department of Life Science, Gwangju Institute of Science and Technology)
Publication Information
Molecules and Cells / v.22, no.2, 2006 , pp. 198-202 More about this Journal
Abstract
The $P2X_7$ receptor, an ATP-gated cation channel, induces cell death in immune cells and is involved in neurodegenerative diseases. Although the receptor plays various roles in these diseases, the cellular mechanisms involved are poorly understood and antagonists are limited. Here, the development of a cell-based assay for human $P2X_7$ receptor is reported. We established permanent lines of HEK 293 cells expressing a high level of $hP2X_7$ receptor. Functional activity of the $hP2X_7$ receptor was confirmed by whole-cell patch recording of ATP-induced ion currents. Prolonged exposure to ATP resulted in death of the $hP2X_7$-expressing HEK 293 cells and this cell death could be quantified. Two known $P2X_7$ antagonists, PPADS and KN-62, blocked ATP-induced death in a concentration-dependent manner. Thus, this assay can be used to screen for new antagonists of $hP2X_7$ receptors.
Keywords
Antagonist; ATP; Cell-based Assay; Cell Death; HEK 293; P2X7 Receptor;
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