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Quantifying Ca2+ current and permeability in ATP-gated P2X7 receptors.


ABSTRACT: ATP-gated P2X7 receptors are prominently expressed in inflammatory cells and play a key role in the immune response. A major consequence of receptor activation is the regulated influx of Ca(2+) through the self-contained cation non-selective channel. Although the physiological importance of the resulting rise in intracellular Ca(2+) is universally acknowledged, the biophysics of the Ca(2+) flux responsible for the effects are poorly understood, largely because traditional methods of measuring Ca(2+) permeability are difficult to apply to P2X7 receptors. Here we use an alternative approach, called dye-overload patch-clamp photometry, to quantify the agonist-gated Ca(2+) flux of recombinant P2X7 receptors of dog, guinea pig, human, monkey, mouse, rat, and zebrafish. We find that the magnitude of the Ca(2+) component of the ATP-gated current depends on the species of origin, the splice variant, and the concentration of the purinergic agonist. We also measured a significant contribution of Ca(2+) to the agonist-gated current of the native P2X7Rs of mouse and human immune cells. Our results provide cross-species quantitative measures of the Ca(2+) current of the P2X7 receptor for the first time, and suggest that the cytoplasmic N terminus plays a meaningful role in regulating the flow of Ca(2+) through the channel.

SUBMITTER: Liang X 

PROVIDER: S-EPMC4367291 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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Quantifying Ca2+ current and permeability in ATP-gated P2X7 receptors.

Liang Xin X   Samways Damien S K DS   Wolf Kyle K   Bowles Elizabeth A EA   Richards Jennifer P JP   Bruno Jonathan J   Dutertre Sébastien S   DiPaolo Richard J RJ   Egan Terrance M TM  

The Journal of biological chemistry 20150202 12


ATP-gated P2X7 receptors are prominently expressed in inflammatory cells and play a key role in the immune response. A major consequence of receptor activation is the regulated influx of Ca(2+) through the self-contained cation non-selective channel. Although the physiological importance of the resulting rise in intracellular Ca(2+) is universally acknowledged, the biophysics of the Ca(2+) flux responsible for the effects are poorly understood, largely because traditional methods of measuring Ca  ...[more]

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