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
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
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were engineered and 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
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were engineered and kindly supplied by Axxam
P2X2/3 recorded from CHO cells on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). 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
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
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
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were engineered and kindly supplied by Axxam
P2X2/3 recorded from CHO cells on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). 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
P2X3 - "Activation and inhibition of P2X3 channels recorded on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note: 
(1.5 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X3 - "Activation and inhibition of P2X3 channels recorded on the Patchliner"
Patchliner application note: (0.6 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X2 / P2X3 - "Pharmacology of P2X2/3 channels recorded on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note: (5 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X2 / P2X3 - "P2X2/3 receptors recorded on Nanion's Patchliner"
Patchliner application note: (0.4 MB)
Cells were kindly provided by Evotec.
Posters
2020 - Kinetic and pharmacological properties of P2X3 and P2X2/3 receptors
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384 instrument) and 
Patchliner poster, 64th Annual Meeting of the Biophysical Society (1.8 MB)
Publications
2019 - Action of MK‐7264 (Gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation
Port-a-Patch publication in British Journal of Pharmacology (2019)
Authors:
Richards D., Gever J.R., Ford A.P., Fountain S.J.
2011 - State-of-the-art automated patch clamp devices: heat activation, action potentials, and high throughput in ion channel screening
Port-a-Patch, Patchliner and 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.
Webinars
28.04.2020 | Webinar: Validation and optimization of automated patch clamp voltage-gated Ca2+ channel assays
Patchliner Webinar
Date: April 28. 2020, 4:00 PM CET (10:00 AM EDT)
Marc will outline the development, optimization and validation of a range of voltage-gated Ca2+ channel assays on the Patchliner automated patch clamp platform that were subsequently used in an 8 year drug discovery collaboration between Metrion Biosciences and a german pharma company.
Speakers:
Dr. Marc Rogers (Chief Scientific Officer, Metrion Biosciences)
Dr. András Horváth (Application Scientist, Nanion Technologies)