Ethanol is a Fast Channel Inhibitor of P2X4 Receptors

Overview

Journal J Pharmacol Exp Ther

Specialty Pharmacology

Date 2011 Jan 8

PMID 21212160

Citations 27

Authors

Olga Ostrovskaya

Liana Asatryan

Letisha Wyatt

Maya Popova

Kaixun Li

Robert W Peoples

Ronald L Alkana

Daryl L Davies

Affiliations

Soon will be listed here.

Abstract

P2X receptors (P2XRs) are ion channels gated by synaptically released ATP. The P2X4 is the most abundant P2XR subtype expressed in the central nervous system and to date is the most ethanol-sensitive. In addition, genomic findings suggest that P2X4Rs may play a role in alcohol intake/preference. However, little is known regarding how ethanol causes the inhibition of ATP-gated currents in P2X4Rs. We begin to address this issue by investigating the effects of ethanol in wild-type and mutant D331A and M336A P2X4Rs expressed in human embryonic kidney (HEK) 293 cells using whole-cell patch-clamp methods. The results suggest that residues D331 and M336 play a role in P2X4R gating and ethanol inhibits channel functioning via a mechanism different from that in other P2XRs. Key findings from the study include: 1) ethanol inhibits ATP-gated currents in a rapid manner; 2) ethanol inhibition of ATP-gated currents does not depend on voltage and ATP concentration; 3) residues 331 and 336 slow P2X4 current deactivation and regulate the inhibitory effects of ethanol; and 4) ethanol effects are similar in HEK293 cells transfected with P2X4Rs and cultured rat hippocampal neurons transduced with P2X4Rs using a recombinant lentiviral system. Overall, these findings provide key information regarding the mechanism of ethanol action on ATP-gated currents in P2X4Rs and provide new insights into the biophysical properties of P2X4Rs.

Citing Articles

An integrated multi-omics analysis of sleep-disordered breathing traits implicates P2XR4 purinergic signaling.

Kurniansyah N, Wallace D, Zhang Y, Yu B, Cade B, Wang H Commun Biol. 2023; 6(1):125.

PMID: 36721044 PMC: 9889381. DOI: 10.1038/s42003-023-04520-y.

Short- and long-term administration of buprenorphine improved gene expression of PX and GABAA receptors in the hippocampus of methamphetamine rats.

Roshani S, Hatami Nemati H, Sadeghian R, Khoshsirat H Heliyon. 2022; 8(11):e11432.

PMID: 36444255 PMC: 9699974. DOI: 10.1016/j.heliyon.2022.e11432.

The P2X4 Receptor: Cellular and Molecular Characteristics of a Promising Neuroinflammatory Target.

Sophocleous R, Ooi L, Sluyter R Int J Mol Sci. 2022; 23(10).

PMID: 35628550 PMC: 9147237. DOI: 10.3390/ijms23105739.

Structural and Functional Features of the P2X4 Receptor: An Immunological Perspective.

Kanellopoulos J, Almeida-da-Silva C, Ruutel Boudinot S, Ojcius D Front Immunol. 2021; 12:645834.

PMID: 33897694 PMC: 8059410. DOI: 10.3389/fimmu.2021.645834.

Implication of Neuronal Versus Microglial P2X4 Receptors in Central Nervous System Disorders.

Duveau A, Bertin E, Boue-Grabot E Neurosci Bull. 2020; 36(11):1327-1343.

PMID: 32889635 PMC: 7674530. DOI: 10.1007/s12264-020-00570-y.

References

Cunha R, Ribeiro J . ATP as a presynaptic modulator. Life Sci. 2001; 68(2):119-37. DOI: 10.1016/s0024-3205(00)00923-1. View

Cheng M, Coalson R, Cascio M . Molecular dynamics simulations of ethanol binding to the transmembrane domain of the glycine receptor: implications for the channel potentiation mechanism. Proteins. 2007; 71(2):972-81. DOI: 10.1002/prot.21784. View

Li C, Peoples R, Weight F . Alcohol action on a neuronal membrane receptor: evidence for a direct interaction with the receptor protein. Proc Natl Acad Sci U S A. 1994; 91(17):8200-4. PMC: 44573. DOI: 10.1073/pnas.91.17.8200. View

Franks N, Lieb W . Molecular and cellular mechanisms of general anaesthesia. Nature. 1994; 367(6464):607-14. DOI: 10.1038/367607a0. View

Lovinger D . Developmental decrease in ethanol inhibition of N-methyl-D-aspartate receptors in rat neocortical neurons: relation to the actions of ifenprodil. J Pharmacol Exp Ther. 1995; 274(1):164-72. View

Khakh B . Molecular physiology of P2X receptors and ATP signalling at synapses. Nat Rev Neurosci. 2001; 2(3):165-74. DOI: 10.1038/35058521. View

Tabakoff B, Saba L, Printz M, Flodman P, Hodgkinson C, Goldman D . Genetical genomic determinants of alcohol consumption in rats and humans. BMC Biol. 2009; 7:70. PMC: 2777866. DOI: 10.1186/1741-7007-7-70. View

Wirkner K, Eberts C, Poelchen W, Allgaier C, Illes P . Mechanism of inhibition by ethanol of NMDA and AMPA receptor channel functions in cultured rat cortical neurons. Naunyn Schmiedebergs Arch Pharmacol. 2001; 362(6):568-76. DOI: 10.1007/s002100000262. View

Soto F, Garcia-Guzman M, Gomez-Hernandez J, Hollmann M, Karschin C, Stuhmer W . P2X4: an ATP-activated ionotropic receptor cloned from rat brain. Proc Natl Acad Sci U S A. 1996; 93(8):3684-8. PMC: 39672. DOI: 10.1073/pnas.93.8.3684. View

10.

Ren H, Honse Y, Peoples R . A site of alcohol action in the fourth membrane-associated domain of the N-methyl-D-aspartate receptor. J Biol Chem. 2003; 278(49):48815-20. DOI: 10.1074/jbc.M302097200. View

11.

Asatryan L, Popova M, Woodward J, King B, Alkana R, Davies D . Roles of ectodomain and transmembrane regions in ethanol and agonist action in purinergic P2X2 and P2X3 receptors. Neuropharmacology. 2008; 55(5):835-43. PMC: 2629788. DOI: 10.1016/j.neuropharm.2008.06.044. View

12.

Jelinkova I, Yan Z, Liang Z, Moonat S, Teisinger J, Stojilkovic S . Identification of P2X4 receptor-specific residues contributing to the ivermectin effects on channel deactivation. Biochem Biophys Res Commun. 2006; 349(2):619-25. DOI: 10.1016/j.bbrc.2006.08.084. View

13.

Peoples R, Weight F . Cutoff in potency implicates alcohol inhibition of N-methyl-D-aspartate receptors in alcohol intoxication. Proc Natl Acad Sci U S A. 1995; 92(7):2825-9. PMC: 42311. DOI: 10.1073/pnas.92.7.2825. View

14.

Li C, Peoples R, Weight F . Ethanol-induced inhibition of a neuronal P2X purinoceptor by an allosteric mechanism. Br J Pharmacol. 1998; 123(1):1-3. PMC: 1565146. DOI: 10.1038/sj.bjp.0701599. View

15.

Yu D, Zhang L, Eisele J, Bertrand D, Changeux J, Weight F . Ethanol inhibition of nicotinic acetylcholine type alpha 7 receptors involves the amino-terminal domain of the receptor. Mol Pharmacol. 1996; 50(4):1010-6. View

16.

Jasti J, Furukawa H, Gonzales E, Gouaux E . Structure of acid-sensing ion channel 1 at 1.9 A resolution and low pH. Nature. 2007; 449(7160):316-23. DOI: 10.1038/nature06163. View

17.

Kobayashi K, Fukuoka T, Yamanaka H, Dai Y, Obata K, Tokunaga A . Differential expression patterns of mRNAs for P2X receptor subunits in neurochemically characterized dorsal root ganglion neurons in the rat. J Comp Neurol. 2004; 481(4):377-90. DOI: 10.1002/cne.20393. View

18.

Colquhoun D . Binding, gating, affinity and efficacy: the interpretation of structure-activity relationships for agonists and of the effects of mutating receptors. Br J Pharmacol. 1998; 125(5):924-47. PMC: 1565672. DOI: 10.1038/sj.bjp.0702164. View

19.

Xiong K, Peoples R, Montgomery J, Chiang Y, Stewart R, Weight F . Differential modulation by copper and zinc of P2X2 and P2X4 receptor function. J Neurophysiol. 1999; 81(5):2088-94. DOI: 10.1152/jn.1999.81.5.2088. View

20.

Torres G, Egan T, Voigt M . Hetero-oligomeric assembly of P2X receptor subunits. Specificities exist with regard to possible partners. J Biol Chem. 1999; 274(10):6653-9. DOI: 10.1074/jbc.274.10.6653. View