Establishment of an assay for P2X7 receptor-mediated cell death.
The P2X7 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 P2X7 receptor is reported. We established permanent lines of HEK 293 cells expressing a high level of hP2X7 receptor. Functional activity of the hP2X7 receptor was confirmed by whole-cell patch recording of ATP-induced ion currents. Prolonged exposure to ATP resulted in death of the hP2X7-expressing HEK 293 cells and this cell death could be quantified. Two known P2X7 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 hP2X7 receptors.
Lee
Song-Yi
SY
Department of Life Science, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea.
Jo
Sooyeon
S
Lee
Ga Eun
GE
Jeong
Lak Shin
LS
Kim
Yong-Chul
YC
Park
Chul-Seung
CS
D000255:Adenosine Triphosphate / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D016923:Cell Death; D002460:Cell Line; D002470:Cell Survival; D006801:Humans; D018408:Patch-Clamp Techniques; D058919:Purinergic P2 Receptor Antagonists; D058486:Receptors, Purinergic P2X7; D014162:Transfection
C546468:P2RX7 protein, human; D058919:Purinergic P2 Receptor Antagonists; D058486:Receptors, Purinergic P2X7; D000255:Adenosine Triphosphate