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

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

Patchliner data and applications:
Cells were kindly supplied by Evotec AG, Hamburg, Germany

P2X_2/3 recorded from 1321N1 cells on the Patchliner. P2X_2/3 was activated repetitively (7 times) by 30 µM ATP.

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

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) data and applications:
Cells were kindly supplied by Axxam

Inhibition of P2X_2/3 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

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) data and applications:
Cells were kindly supplied by Axxam

P2X_2/3 recorded from CHO cells on the SyncroPatch 384PE. P2X_2/3 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

Patchliner data and applications:
Cells were kindly supplied by Evotec AG, Hamburg, Germany

Block of P2X_2/3 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

Patchliner data and applications:
Cells were kindly supplied by Evotec AG, Hamburg, Germany

P2X_2/3 recorded from 1321N1 cells on the Patchliner. P2X_2/3 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

SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) data and applications:
Cells were kindly supplied by Axxam

P2X_2/3 recorded from CHO cells on the SyncroPatch 384PE. P2X_2/3 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.