• P2X3

    Reproducable current traces from 1321 N1 cells expressing P2X3 receptors, activated by 100 ms application of 10 µM ATP (measured on the Port-a-Patch).

P2X3 | P2X Purinoceptor 3

Family:
Purinergic Receptors P2X

Subgroups:
The P2X family belongs to the P2 receptors superfamily which has further been divided into five subclasses: P2X, P2Y, P2Z, P2U, and P2T. The ligand-gated purinergic receptor class P2X contains seven members (P2X1 - P2X7).

Topology:
One molecule is composed of only two transmembrane domains and contains a large extracellular loop and intracellular carboxyl and amino termini

Regulation and Function:
P2X receptors are ligand-gated ion channels and are activated by extracellular ATP. Greatly distributed in neurons and glial cells, P2X receptors mediate a large variety of responses including fast transmission at central synapses, contraction of smooth muscle cells, platelet aggregation, macrophage activation, and apoptosis.

Data and Applications

P2X3 - Application of ATP

PortiP2X3icon pap   Port-a-Patch data and applications:
Cells were kindly supplied by Evotec AG, Hamburg, Germany

Shown are raw current traces from 1321 N1 cells expressing P2X3 receptors demonstrating the reproducability of the whole cell responses. Currents were activated by 100 ms application of 10 µM ATP. The waiting time between sweeps was 120 s (holding −60 mV).

P2X2/ P2X3 - Repetitive activation on the Patchliner

P2X2 slide 1icon pl   Patchliner data and applications: 
Cells were kindly supplied by Evotec AG, Hamburg, Germany

P2X2/3 recorded from 1321N1 cells on the Patchliner. P2X2/3 was activated repetitively (7 times) by 30 µM ATP.

P2X2/ P2X3 - High throughput screening on the SyncroPatch 384PE

P2X23 CHO Cells Suramin SP384 Screenshoticon sp96   SyncroPatch 384PE data and applications: 
Cells were kindly supplied by Axxam

Inhibition of P2X2/3 by suramin. Suramin was pre-incubated at each concentration for 3 - 5 mins followed by co-application with 10 µM ATP. All 3 concentrations of suramin were applied to each well and a cumulative concentration response curve was constructed for n = 372 wells out of a possible 384. Success rate was 97% for completed experiments. .

P2X2/ P2X3 - Block by suramin on the SyncroPatch 384PE

P2X23 CHO Cells suramin SP384icon sp96   SyncroPatch 384PE data and applications: 
Cells were kindly supplied by Axxam

P2X2/3 recorded from CHO cells on the SyncroPatch 384PE. P2X2/3 was activated by 10 µM ATP and blocked by increasing concentrations of suramin with an IC50 = 17.7 ± 0.9 µM (n = 372). Average traces and concentration response curve shown for 372 cells. Success rate was 97% for completed experiment (372 wells from a possible 384). Multi-hole (4 holes per well) chips were used.

P2X2/ P2X3 - Block by suramin on the Patchliner

P2X23 1321N1Cells suramin PLicon pl   Patchliner data and applications: 
Cells were kindly supplied by Evotec AG, Hamburg, Germany

Block of P2X2/3 receptors by suramin. Suramin at increasing concentrations (1 μM - 1 mM) was pre-incubated and then co-applied with 30 μM ATP. Full recovery from block was achieved upon washout (data not shown). Concentration response curve (right) for suramin block, IC50 = 28.0 ± 5.3 μM (n = 7).

P2X2/ P2X3 - Application of ATP on the Patchliner

P2X23 1321N1Cells ATP PLicon pl   Patchliner data and applications: 
Cells were kindly supplied by Evotec AG, Hamburg, Germany

P2X2/3 recorded from 1321N1 cells on the Patchliner. P2X2/3 was activated by increasing concentrations of ATP with an EC50 = 7.8 ± 1.0 µM (n = 10). ATP was applied for approximately 600 ms using the stacked solutions function of the Patchliner (holding potential = -80mV).

P2X2/ P2X3 - Activation by ATP on the SyncroPatch 384PE

P2X23 CHOCells ATP SP384icon sp96   SyncroPatch 384PE data and applications: 
Cells were kindly supplied by Axxam

P2X2/3 recorded from CHO cells on the SyncroPatch 384PE. P2X2/3 was activated by increasing concentrations of ATP with an EC50 = 1.1 µM (n = 191). ATP was applied for approximately 1.5 s using the ‘Ligand Puff’ function of the SyncroPatch 384PE. Average traces and concentration response curve shown for 191 cells. Success rate was 99% for completed experiment (191 wells from a possible 192). Multi-hole (4 holes per well) chips were used.

Application Notes

P2X2 / P2X3 - "Pharmacology of P2X2/3 channels recorded on the SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE application note:   logo pdf   (5 MB)
Cells were kindly provided by Axxam S.p.A., Milan.  

P2X2 / P2X3 - "P2X2/3 receptors recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Evotec.  

Publications

2011 - State-of-the-art automated patch clamp devices: heat activation, action potentials, and high throughput in ion channel screening

icon pap   Port-a-Patch,   icon pl  Patchliner and   icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Frontiers in Pharmacology (2011)

Authors: 
Stoelzle S., Obergrussberger A., Brüggemann A., Haarmann C., George M., Kettenhofen R., Fertig N.

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